ORCID Profile
0000-0002-4341-3265
Current Organisations
Zist Pishro Co
,
University of Melbourne
,
University of Adelaide
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Publisher: ZappyLab, Inc.
Date: 03-05-2020
DOI: 10.17504/PROTOCOLS.IO.BFWXJPFN
Abstract: A pelvic nerve array is custom designed for implantation onto the pelvic nerve of male rats. This device is used for long-term (recovery) experiments. The device can electrically stimulate the pelvic nerve as well as record electrically evoked or spontaneous neural activity. Previously, we have successfully implanted rats for 2 months with no reports of biological adverse complications or damage sustained to the array. The surgical procedure is performed under anesthesia and should incorporate all local requirements for standards of animal experimentation, including methods of anesthesia, surgical environment, and post-operative monitoring and care.
Publisher: Bioscientifica
Date: 15-05-2013
DOI: 10.1530/JOE-13-0056
Abstract: Neurturin (NTN) is a member of the glial cell line-derived neurotrophic factor (GDNF) family and signals through GDNF family receptor alpha 2 (GFRα2). We hypothesised that epithelial atrophy reported in the reproductive organs of Ntn ( Nrtn )- and Gfr α 2 ( Gfra2 )-deficient mice could be due to NTN affecting the hormonal environment. To investigate this, we compared the reproductive organs of Ntn - and Gfr α 2 -deficient male mice in parallel with an analysis of their circulating reproductive hormone levels. There were no significant structural changes within the organs of the knockout mice however, serum and intratesticular testosterone and serum LH levels were very low. To reconcile these observations, we tested androgen sensitivity by creating a dihydrotestosterone (DHT) cl (castration plus DHT implant) to create fixed circulating levels of androgens, allowing the evaluation of androgen-sensitive endpoints. At the same serum DHT levels, serum LH levels were lower and prostate and seminal vesicle weights were higher in the Ntn knockout (NTNKO) mice than in the wild-type mice, suggesting an increased response to androgens in the accessory glands and hypothalamus and pituitary of the NTNKO mice. Testicular and pituitary responsiveness was unaffected in the NTNKO males, as determined by the response to the human chorionic gonadotrophin or GNRH analogue, leuprolide, respectively. In conclusion, our results suggest that NTN inactivation enhances androgen sensitivity in reproductive and neuroendocrine tissues, revealing a novel mechanism to influence reproductive function and the activity of other androgen-dependent tissues.
Publisher: Elsevier
Date: 2006
Publisher: Wiley
Date: 19-11-2015
DOI: 10.1111/ASJ.12549
Abstract: The study examined the effects of blend of 80% canola oil and 20% palm oil (BCPO) on nutrient intake and digestibility, growth performance, rumen fermentation and fatty acids (FA) in goats. Twenty-four Boer bucks were randomly assigned to diets containing 0, 4 and 8% BCPO on a dry matter basis, fed for 100 days and slaughtered. Diet did not affect feed efficiency, growth performance, intake and digestibility of all nutrients except ether extract. Intakes and digestibilities of ether extract, unsaturated fatty acids (FA) and total FA were higher (P < 0.05) while digestibility of C18:0 was lower (P < 0.05) in oil-fed goats than the control goats. Total volatile FA, acetate, butyrate, acetate ropionate ratio and methane decreased (P < 0.05) with increasing BCPO but propionate, NH3 -N and rumen pH did not differ between diets. Ruminal concentration of C18:0, n-3 FA and total FA increased (P < 0.05) while C12:0, C14:0, C15:0 and n-6 FA decreased with increasing BCPO. Analysis of the FA composition of Triceps brachii muscle showed that concentrations of C16:0, C14:0 and C18:2n-6 were lower (P < 0.05) while C18:1n-9, C18:3n-3 and C20:5n-3 were higher in oil-fed goats compared with control goats. Dietary BCPO altered muscle lipids without having detrimental effects on nutrient intake and digestibility and growth performance in goats.
Publisher: Istanbul University
Date: 04-01-2016
Publisher: Wiley
Date: 05-11-2014
DOI: 10.1111/ASJ.12279
Abstract: The study appraised the effects of Carotino oil on in vitro rumen fermentation, gas production, metabolism and apparent biohydrogenation of oleic, linoleic and linolenic acids. Carotino oil was added to a basal diet (50% concentrate and 50% oil palm frond) at the rate of 0, 2, 4, 6 and 8% dry matter of the diet. Rumen inoculum was obtained from three fistulated Boer bucks and incubated with 200 mg of each treatment for 24 h at 39°C. Gas production, fermentation kinetics, in vitro organic matter digestibility (IVOMD), volatile fatty acids (VFA), in vitro dry matter digestibility (IVDMD), metabolizable energy and free fatty acids were determined. Carotino oil did not affect (P > 0.05) gas production, metabolizable energy, pH, IVOMD, IVDMD, methane, total and in idual VFAs. However, Carotino oil decreased (P < 0.05) the biohydrogenation of linoleic and linolenic acids but enhanced (P < 0.05) the biohydrogenation of oleic acid. After 24 h incubation, the concentrations of stearic, palmitic, pentadecanoic, myristic, myristoleic and lauric acids decreased (P < 0.05) while the concentration of linolenic, linoleic, oleic and transvaccenic acids and conjugated linoleic acid (CLAc9t11) increased (P < 0.05) with increasing levels of Carotino oil. Carotino oil seems to enhance the accumulation of beneficial unsaturated fatty acids without disrupting rumen fermentation.
Publisher: Elsevier BV
Date: 05-2016
Publisher: Wiley
Date: 15-10-1989
Abstract: Retrograde-tracing and immunohistochemical techniques were used in combination to investigate the types of putative transmitters in pelvic neurons that project to the bladder, colon or penis of rats. In addition, populations of axon varicosities associated with these neurons were characterized. Subpopulations of neurons in colchicine-treated major pelvic ganglia and accessory ganglia of male rats contained immunoreactivity (IR) for tyrosine hydroxylase (TH), vasoactive intestinal peptide (VIP), neuropeptide Y (NPY), or enkephalin (ENK), while types of immunoreactivity found in major groups of varicose axons were ENK, cholecystokinin (CCK), and somatostatin (SOM). Substance P (SP)-IR varicose axons were much less common. Bladder and colon neurons were similar in a number of ways. Many neurons contained NPY-IR (greater than or equal to 50%), fewer contained TH-IR (25-30%), and even fewer contained ENK-IR (5-15%) or VIP-IR (5-10%) many neurons were associated with baskets of ENK-IR varicosities (50-65%) and fewer neurons were surrounded by CCK- or SOM-IR varicosities (30-35%). Colon neurons differed from penis neurons in having a slightly larger proportion that contained ENK-IR (10-15%, compared with 1-3%). Penis neurons were markedly different from the other two groups in additional ways. More than 90% of them contained VIP-IR, whereas only 5-7% contained NPY-IR and none were immunoreactive for TH. Furthermore, although the proportion of penile neurons associated with many ENK-IR varicosities was similar to the bladder and colon neurons (45-50%), they were rarely seen close to CCK- or SOM-IR varicose axons. These studies describe similarities and differences in the histochemical properties of neurons which project to the bladder, colon, or penis and of the varicose axons associated with those neurons. This gives further insights into the possible transmitter mechanisms involved in the regulation of different pelvic functions.
Publisher: Elsevier BV
Date: 08-1994
Abstract: The structural and functional unit of the thyroid gland is the follicle, consisting of a closed lumen surrounded by a single layer of polarized epithelial cells. In this paper we have attempted to characterize the process of lumenal development when primary cultures of porcine thyroid cells reorganized to form follicles. Cells incubated with the loop diuretic, bumetanide, an inhibitor of NaK2Cl cotransport, aggregated but failed to form normal follicles. Laser scanning confocal microscopy combined with immunohistochemical markers of thyroid cell-surface proteins demonstrated that in the presence of bumetanide cells polarized and assembled ZO-1-containing tight junctions separating their apical and basolateral membrane domains. Cultures formed small lumena but their subsequent growth was inhibited by bumetanide. Electrophysiological studies confirmed that bumetanide-sensitive Cl- transport was the major contributor to the transepithelial electrical potential difference across the follicular wall after 48 h incubation. Other potential mechanisms did not contribute significantly to follicular lumenal growth. In particular, bumetanide did not affect cell proliferation and, in contrast to tissue follicles, thyroglobulin could not be detected within the lumena of cultured follicles. We conclude that thyroid follicular reorganization involves two distinct and separate phases of lumenal development: initial lumen formation which probably reflects the assembly of a specialized apical membrane domain and subsequent lumenal growth which is mediated by the inward transport of Cl- by polarized epithelial cells.
Publisher: Public Library of Science (PLoS)
Date: 05-07-2018
Publisher: Frontiers Media SA
Date: 2011
Publisher: Elsevier BV
Date: 09-2007
DOI: 10.1016/J.EXPNEUROL.2007.05.011
Abstract: Penile (cavernous) nerves are readily damaged during radical prostatectomy, invariably causing impotence. Erectile function can return, however this may take months or years and capacity often remains poor. Many studies have attempted to improve penile nerve regeneration but have not explored mechanisms underlying the delay in functional recovery. This is assumed to be due to slow growth of axons, although penile tissues also change following loss of erectile activity. We have asked whether delayed recovery of the nitrergic nerve-evoked erectile response is due to pre-synaptic (slow axonal growth) or post-synaptic (changes in tissue responsiveness) mechanisms. These components were dissected in vitro following penile nerve injury in adult rats. Following crush of both penile nerves, excellent regeneration of nitrergic axons occurred after 10-12 weeks but neurogenic relaxation of cavernosum muscle was still relatively poor. This was at least partly due to attenuated tissue responsiveness to nitric oxide (using sodium nitroprusside as a donor) from 3 weeks after injury. Western blotting also revealed a modest reduction of soluble guanylyl cyclase. A second model of penile nerve injury, unilateral cut, completely denervated one side but retained potential for penile erection. Some anatomical and functional recovery occurred after 9-11 weeks (probably due to sprouting from contralateral uninjured axons), but nitroprusside-evoked relaxations were unaltered from at least 3 weeks onward. These data suggest that erectile dysfunction following extensive nerve injury may be exacerbated by postsynaptic changes in nitric oxide signaling, even when nerve regeneration occurs. This may be prevented by continued activation of penile tissues to retain normal perfusion.
Publisher: ZappyLab, Inc.
Date: 17-12-2019
DOI: 10.17504/PROTOCOLS.IO.BAKMICU6
Abstract: This protocol is used for analysing expression pattens of immediate early gene products (e.g., c-Fos) in immunostained transverse sections of spinal cord.
Publisher: ZappyLab, Inc.
Date: 17-12-2019
DOI: 10.17504/PROTOCOLS.IO.BAKKICUW
Abstract: This protocol is used for immunohistochemical visualisation of immediate early gene expression (c-Fos or Egr-1) in cryosections of rat lumbosacral spinal cord. Free-floating sections are processed in a double labelling protocol to distinguish immediate early gene expression in different neurochemical classes of spinal cord neurons: ChAT [choline acetyltransferase]: preganglionic neurons TH [tyrosine hydroxyls]: dopaminergic neurons Pax2: inhibitory interneurons
Publisher: Inter-Research Science Center
Date: 09-08-2017
DOI: 10.3354/AB00678
Publisher: Medwell Publications
Date: 12-2012
Publisher: Society for Neuroscience
Date: 11-2021
DOI: 10.1523/ENEURO.0364-21.2021
Abstract: Sensorimotor circuits of the lumbosacral spinal cord are required for lower urinary tract (LUT) regulation as well as being engaged in pelvic pain states. To date, no molecular markers have been identified to enable specific visualization of LUT afferents, which are embedded within spinal cord segments that also subserve somatic functions. Moreover, previous studies have not fully investigated the patterning within or across spinal segments, compared afferent innervation of the bladder and urethra, or explored possible structural sex differences in these pathways. We have addressed these questions in adult Sprague Dawley rats, using intramural microinjection of the tract tracer, B subunit of cholera toxin (CTB). Afferent distribution was analyzed within in idual sections and 3D reconstructions from sections across four spinal cord segments (L5-S2), and in cleared intact spinal cord viewed with light sheet microscopy. Simultaneous mapping of preganglionic neurons showed their location throughout S1 but restricted to the caudal half of L6. Afferents from both LUT regions extended from L5 to S2, even where preganglionic motor pathways were absent. In L6 and S1, most afferents were associated with the sacral preganglionic nucleus (SPN) and sacral dorsal commissural nucleus (SDCom), with very few in the superficial laminae of the dorsal horn. Spinal innervation patterns by bladder and urethra afferents were remarkably similar, likewise the patterning in male and female rats. In conclusion, microscale to macroscale mapping has identified distinct features of LUT afferent projections to the lumbosacral cord and provided a new anatomic approach for future studies on plasticity, injury responses, and modeling of these pathways.
Publisher: Wiley
Date: 17-12-2008
DOI: 10.1002/CNE.21535
Publisher: Istanbul University
Date: 04-07-2016
Publisher: Springer Science and Business Media LLC
Date: 08-02-2016
Publisher: Wiley
Date: 28-01-2011
Publisher: MDPI AG
Date: 13-07-2015
Publisher: Wiley
Date: 05-2017
Publisher: Wiley
Date: 19-07-2007
DOI: 10.1002/CNE.21412
Abstract: Sprouting of peptidergic nociceptive and descending supraspinal projections to the dorsal horn following spinal cord injury (SCI) has been proposed as a mechanism of neuropathic pain. To identify structural changes that could initiate or maintain SCI pain, we used a complete transection model in rats to examine how structural remodeling in the dorsal horn rostral to the lesion relates to distance from injury, laminar region, and duration of injury. The major classes of C-fiber primary afferents differed greatly in their susceptibility to structural and chemical changes and their ability to undergo plasticity. Peptidergic primary afferents showed a widespread loss throughout the dorsal horn of segments approaching the injury site. Some of this loss may have been due to decreased neuropeptide expression. The reduction in peptidergic fibers was transient, indicating compensatory sprouting and perhaps also increased neuropeptide expression within the cord. Nonpeptidergic afferents expressing GFRalpha1 were largely unaffected by SCI. In contrast, in GFRalpha2-expressing nonpeptidergic afferents SCI caused a permanent loss of dorsal horn innervation. Unexpectedly, GFRalpha2 was transiently induced throughout deeper laminae but this was not due to upregulation of GFRalpha2 in dorsal root ganglia. We also observed permanent sprouting of catecholamine terminals of supraspinal origin. This was restricted to the superficial laminae. Our results show that SCI caused a loss of sensory input as well as structural remodeling such that the balance of nociceptive inputs and descending modulation was permanently altered. These changes may contribute to mechanisms rostral to the site of SCI that trigger and maintain neuropathic pain.
Publisher: Elsevier BV
Date: 05-2003
Publisher: Elsevier BV
Date: 08-2021
Publisher: Springer Science and Business Media LLC
Date: 02-10-2014
Publisher: No publisher found
Date: 1998
DOI: 10.1002/(SICI)1096-9861(19980713)396:4<429::AID-CNE2>3.0.CO;2-3 [PII]
Publisher: Wiley
Date: 08-07-1985
Abstract: The distributions of peptide-containing nerve fibers and cell bodies in the canine small intestine were determined with antibodies raised against seven peptides: enkephalin, gastrin-releasing peptide (GRP), neuropeptide Y, neurotensin, somatostatin, substance P, and vasoactive intestinal peptide (VIP). Immunoreactive nerve cell bodies and fibers were found for each peptide except neurotensin. In the muscle layers there were numerous substance P, VIP, and enkephalin fibers, fewer neuropeptide Y fibers, and very few GRP or somatostatin fibers. The mucosa contained many VIP and substance P fibers, moderate numbers of neuropeptide Y, somatostatin, and GRP fibers and rare enkephalin fibers. Nerve cell bodies reactive for each of the six neural peptides were located in both the myenteric and submucous plexuses. The distributions of nerve cell bodies and processes in the canine small intestine show many similarities with other mammals, for ex le, in the distributions of VIP, substance P, neuropeptide Y, and somatostatin nerves. There are some major differences, such as the presence in dogs of numerous submucosal nerve cell bodies with enkephalinlike immunoreactivity and of GRP-like immunoreactivity in submucous nerve cell bodies and mucosal fibers.
Publisher: Wiley
Date: 2000
DOI: 10.1002/1096-9861(20000904)424:4<577::AID-CNE2>3.0.CO;2-E
Publisher: Elsevier BV
Date: 06-2004
Publisher: The Royal Society
Date: 22-04-1995
Abstract: Platypus mucous gland electroreceptors differ from electroreceptors described for fish in that they lack an associated specialized sensory cell. Thus a bare nerve terminal is used to detect electrical stimuli, and also to generate local and action potentials. Previous studies have identified these terminals (an average of 16 per mucous gland), but had not shown whether the terminals have direct contact with the duct of the mucous gland. This poses the problem of how the electrical stimulus reaches the nerve terminals. This study demonstrates the portions of the nerve terminals responsible for electroreception, and shows how these portions use the surrounding epidermal tissue to overcome the combined problems of lacking a sensory cell and making physical contact with the conducting medium in the duct of the gland. A terminal axonal filament is described which accommodates for these problems, the terminal filament provides a low-resistance pathway for the electrical stimuli, and is embedded with its proximal and distal portions in high and low resistance epidermis, respectively. Lateral interactions occur between adjacent terminal filaments via a plexus that is directed circumferentially around the duct from the proximal portion of the terminal filament. These circumferential arbors form an interconnecting ring between all 16 terminal filaments, and may be used to lower the signal-to-noise ratio of the electroreceptor and thus enhance overall sensitivity.
Publisher: Elsevier BV
Date: 09-2017
Publisher: Elsevier BV
Date: 07-2022
Publisher: Wiley
Date: 1995
DOI: 10.1002/CM.970320408
Abstract: In epithelial cells interactions between the actin cytoskeleton and cell-cell junctions regulate paracellular permeability and participate in morphogenesis. We have studied the relationship between supracellular morphology and actin-junction interactions using primary cultures of porcine thyroid cells grown either as three-dimensional follicles or as open monolayers. Regardless of morphology, thyroid cells assembled occluding and adhesive junctions containing ZO-1 and E-cadherin, respectively, and showed F-actin staining in apical microvilli and a perijunctional ring. In monolayers, actin stress fibers were also observed in the apical and basal poles of cells, where they terminated in the vinculin-rich zonula adherens and in cell-substrate focal adhesions, respectively. Surprisingly, we were unable to detect vinculin localization in follicular cells, which also did not form stress fibers. Immunoblotting confirmed significantly greater vinculin in triton-insoluble fractions from monolayer cells compared with follicular cells. Incubation of monolayers with 8 chloro(phenylthio)-cyclic AMP decreased the level of immunodetectable vinculin in the zonula adherens, indicating that junctional incorporation of vinculin was regulated by cyclic AMP. In monolayer cultures, cytochalasin D (1 microM) cause actin filaments to aggregate associated with retraction of cells from one another and the disruption of cell junctions. Despite morphologically similar perturbations of actin organization in follicular cultures treated with cytochalasin D, junctional staining of ZO-1 and E-cadherin was preserved and cells remained adherent to one another. We conclude that in cultured thyroid cells structural and functional associations between actin filaments and cellular junctions differ depending upon the supracellular morphology in which cells are grown. One important underlying mechanism appears to be regulation of vinculin incorporation into adhesive junctions by cyclic AMP.
Publisher: Springer Science and Business Media LLC
Date: 1993
DOI: 10.1038/361310B0
Abstract: Sirtuin 1 (SIRT1) plays a very important role in a wide range of biological responses, such as metabolism, inflammation and cell apoptosis. Changes in the levels of SIRT1 have been detected in the brain after traumatic brain injury (TBI). Further, SIRT1 has shown a neuroprotective effect in some models of neuronal death however, its role and working mechanisms are not well understood in the model of TBI. This study aimed to address this issue. SIRT1-specific inhibitor (sirtinol) and activator (A3) were introduced to explore the role of SIRT1 in cell apoptosis. Results of the study suggest that SIRT1 plays an important role in neuronal apoptosis after TBI by inhibiting NF-κB, IL-6 and TNF-α deacetylation and the apoptotic pathway sequentially, possibly by alleviating neuroinflammation.
Publisher: Wiley
Date: 22-09-2017
DOI: 10.1080/08997659.2017.1374310
Abstract: Aqueous and methanol extracts of lemon Citrus limon peel, Euphorbia hirta (aerial parts), and fenugreek Trigonella foenum-graecum seeds were tested for their in vitro antimicrobial activities against the bacterium Aeromonas hydrophila. A swab paper disk method showed that the methanol extract of E. hirta (EHE) had the largest inhibition zone and the lowest minimal inhibitory concentration compared to all other herbal extracts. Based on these results, EHE was included in the diets of Sharptooth Catfish Clarias gariepinus at 0 (control), 2, 5, or 7 g/kg of diet (experiment 1). Each treatment was conducted in triplicate, with 30 fish (mean weight ± SE = 9.4 ± 0.4 g) in each replicate. After 30 d, the growth, feed intake, hepatosomatic index (HSI), and plasma biochemical parameters were measured. With a separate batch of Sharptooth Catfish, the efficacy of the EHE diets in conferring fish resistance to A. hydrophila over 30 d was compared to that of a diet containing oxytetracycline (OTC experiment 2). Six treatments were conducted in triplicate groups of 30 fish (mean weight ± SE = 9.0 ± 0.3 g) the Control fish were fed the control diet and were not injected with A. hydrophila, while the Control-AH and OTC-AH groups were infected with A. hydrophila and were fed either the control diet or the diet containing OTC at 1 g/199 g. The other three treatments included fish that were injected with A. hydrophila but fed diets with increasing EHE at 2, 5, or 7 g/kg. Experiment 1 showed no change to growth, feeding efficiency, HSI, or plasma biochemical parameters. In experiment 2, however, fish that were fed dietary EHE at 5 g/kg had significantly lower mortality than the Control-AH group, with further resistance observed for fish fed EHE at 7 g/kg. Dietary OTC was more effective than EHE as a prophylactic to A. hydrophila infection in Sharptooth Catfish. Nevertheless, EHE can potentially be a valuable dietary supplement to improve the resistance of Sharptooth Catfish to A. hydrophila infection. Received May 3, 2017 accepted August 24, 2017.
Publisher: Springer Science and Business Media LLC
Date: 02-06-2017
Publisher: Wiley
Date: 29-06-2020
DOI: 10.1111/JOA.13221
Publisher: Wiley
Date: 13-07-1998
DOI: 10.1002/(SICI)1096-9861(19980713)396:4<429::AID-CNE2>3.0.CO;2-3
Abstract: In recent years, considerable progress has been made in characterising the neural circuitry of the pelvic plexus, particularly in the male rat. However, the small ganglia on the adventitial surface of the rectum remain largely unstudied. We have used immunohistochemistry and retrograde tracing techniques to determine the content and projections of these neurons. The adventitial ganglia contain 600-1,000 neurons. All of these are immunoreactive for choline acetyltransferase, 44% are immunoreactive for calbindin, and 35% are immunoreactive for vasoactive intestinal peptide. Very few (1-5%) adventitial neurons contain tyrosine hydroxylase or neuropeptide Y. In contrast, most adventitial neurons are surrounded by varicose axons that do contain tyrosine hydroxylase or neuropeptide Y. Retrograde tracing studies showed that the primary targets of adventitial neurons within the bowel are the internal anal sphincter and the circular muscle directly adjacent to the sphincter. However, more adventitial neurons project out of the gut wall than to targets within the bowel. These are most likely to be viscerofugal and rectospinal neurons. Combining the immunohistochemical and tracing observations, these studies suggest that the rat adventitial ganglia do not represent an additional source of pelvic (autonomic postganglionic) neurons but, instead, that they are comprised primarily of viscerofugal and rectospinal neurons. This is very different from the adventitial rectal ganglia of the cat, which represent merely an extension of the pelvic plexus.
Publisher: Triveni Enterprises
Date: 15-09-2018
Publisher: Frontiers Media SA
Date: 2013
Publisher: Informa UK Limited
Date: 08-2012
DOI: 10.1080/03079457.2012.691155
Abstract: Stressors may influence chicken susceptibility to pathogens such as Salmonella enterica. Feed withdrawal stress can cause changes in normal intestinal epithelial structure and may lead to increased attachment and colonization of Salmonella. This study aimed to investigate modulatory effects of epigenetic modification by feed restriction on S. enterica serovar Enteritidis colonization in broiler chickens subjected to feed withdrawal stress. Chicks were ided into four groups: ad libitum feeding ad libitum feeding with 24-h feed withdrawal on day 42 60% feed restriction on days 4, 5, and 6 and 60% feed restriction on days 4, 5, and 6 with 24-h feed withdrawal on day 42. Attachment of S. Enteritidis to ileal tissue was determined using an ex vivo ileal loop assay, and heat shock protein 70 (Hsp70) expression was evaluated using sodium dodecyl sulphate-polyacrylamide gel electrophoresis and western blotting. Feed withdrawal stress increased S. Enteritidis attachment to ileal tissue. However, following feed withdrawal the epigenetically modified chickens had significantly lower attachment of S. Enteritidis than their control counterparts. A similar trend with a very positive correlation was observed for Hsp70 expression. It appears that epigenetic modification can enhance resistance to S. Enteritidis colonization later in life in chickens under stress conditions. The underlying mechanism could be associated with the lower Hsp70 expression in the epigenetically modified chickens.
Publisher: Elsevier BV
Date: 02-2009
DOI: 10.1016/J.EXPNEUROL.2008.10.012
Abstract: The pelvic ganglia provide autonomic innervation to pelvic viscera and urogenital organs. These neurons are susceptible to axonal injury during pelvic surgical procedures, yet their regenerative mechanisms are poorly understood. The AP-1 transcription factor component, c-Jun, has been strongly linked to regenerative events in injured sensory, sympathetic and somatic motor neurons and is considered to be required for regeneration. Our aims were: (1) to identify whether c-Jun was upregulated by injury in pelvic parasympathetic neurons, and (2) whether injury was required for c-Jun upregulation, by performing deafferentation (i.e., severance of lumbar and sacral spinal inputs), which elicits sprouting of axon collaterals from pelvic ganglion neurons but does not injure them. A week after penile nerve axotomy in rats and mice, upregulation of c-Jun occurred in numerous glia within pelvic ganglia and almost half of the retrogradely-labelled penis-projecting neurons but also occurred in many uninjured noradrenergic neurons. We also identified upregulation of c-Jun in many pelvic ganglion neurons and glia a week after deafferentation, suggesting that c-Jun expression is activated in sprouting but uninjured neurons. A c-Jun response was retained in injured or deafferented parasympathetic neurons in neurturin knockout mice. In summary, neurturin-independent c-Jun expression within pelvic ganglion neurons does not require a direct injury and may instead be causally linked to axonal sprouting, regardless of stimulus. This study revealed mechanisms involved in structural remodelling of pelvic autonomic nerve circuits that may be modulated to improve regenerative processes.
Publisher: Hindawi Limited
Date: 16-08-2018
DOI: 10.1111/ANU.12819
Publisher: Oxford University Press (OUP)
Date: 11-2009
DOI: 10.1111/J.1743-6109.2009.01415.X
Abstract: The metabolic syndrome is a cluster of cardiovascular risk factors that predispose toward the development of diseases such as diabetes. Erectile dysfunction (ED) is common in men with metabolic syndrome, but its etiology is poorly understood. Pro-erectile nitrergic nerves innervating penile erectile tissue are also susceptible to mechanical injury during pelvic surgical procedures, which can lead to sexual dysfunction. The aims of this article are: (i) to examine erectile function in an experimental model of metabolic syndrome, the phosphoenolpyruvate carboxykinase (PEPCK)-overexpressing rat and (ii) to study function and cavernous reinnervation after penile nerve crush injury, which permits regeneration, in transgenic rats. We analyzed the density of noradrenergic and nitrergic nerves and performed organ bath pharmacology to assess neurogenic activity. By analyzing changes in neural structure, function, and pharmacologic responses of cavernous tissue after nerve crush injury, we were able to reveal neurologic deficits in rats with metabolic syndrome. Animals with features of metabolic syndrome did not develop notable changes in cavernous autonomic nerve density or nerve-evoked smooth muscle activity. However, regeneration of nitrergic nerves after crush injury in transgenic rats was impaired compared with injured controls. This was manifested as a deficit in axon regrowth and responses to axon activation. However, unlike injured controls, injured PEPCK-overexpressing rats did not develop a reduced maximal response to the nitric oxide (NO) donor, sodium nitroprusside. This suggests preserved NO responsiveness in tissues from rats with metabolic syndrome, despite impaired regeneration and return of function. This study revealed that rats with features of metabolic syndrome display impaired cavernous nerve regeneration after penile nerve injury, but the degree of functional impairment may be attenuated due to reduced plasticity of NO signaling. This reinnervation deficit may be of clinical relevance for understanding why ED persists in some (particularly aged) men after pelvic surgery.
Publisher: Informa UK Limited
Date: 24-03-2016
Publisher: Springer Science and Business Media LLC
Date: 08-07-2019
Publisher: Oxford University Press (OUP)
Date: 23-06-2021
Publisher: Informa UK Limited
Date: 07-2013
DOI: 10.4161/ORG.25886
Publisher: No publisher found
Date: 2000
DOI: 10.1002/1096-9861(20000904)424:4<577::AID-CNE2>3.0.CO;2-E [PII]
Publisher: ZappyLab, Inc.
Date: 14-01-2019
DOI: 10.17504/PROTOCOLS.IO.W3GFGJW
Abstract: This collection describes the procedures required to label, visualise, characterise and quantify neurons that innervate the lower urinary tract tissues of adult male and female Sprague-Dawley rats. This collection includes protocols for: STAGE 1: Surgery to micro-inject fluorescent retrograde tracer dyes into one or more sites within the lower urinary tract STAGE 2: Intracardiac perfusion with fixative to preserve neural tissues of interest STAGE 3: Fluorescence immunohistochemistry of ganglion cryosections.
Publisher: Springer Science and Business Media LLC
Date: 10-2011
Publisher: Springer Science and Business Media LLC
Date: 08-01-2003
DOI: 10.1007/S00441-002-0673-1
Abstract: Pelvic ganglia are mixed sympathetic-parasympathetic ganglia and provide the majority of the autonomic innervation to the urogenital organs. Here we describe the structural and histochemical features of the major pelvic ganglion in the male mouse and compare two different mouse strains. The basic structural features of the ganglion are similar to those in the male rat. Almost all pelvic ganglion cells are monopolar and most are cholinergic. All contain either neuropeptide Y (NPY) or vasoactive intestinal peptide (VIP), or both peptides together. The peptide coexistence varies between strains, with C57BL/6 mice having similar proportions of neurons with NPY alone, VIP alone or both peptides. In contrast, virtually all pelvic neurons in the Quackenbush-Swiss (QS) strain express NPY, i.e. the level of VIP/NPY coexistence is much higher. Cholinergic axons provide the major nerve supply to epithelia of reproductive organs, bladder smooth muscle and, as described previously, penile erectile tissue. They also provide a minor component of the smooth muscle innervation of the prostate gland, seminal vesicles and vas deferens. Virtually all non-cholinergic pelvic ganglion cells are noradrenergic and contain NPY. Their major target is smooth muscle of reproductive organs. This study shows that the male mouse pelvic ganglion bears many similarities to that in the rat, but that VIP/NPY colocalisation is much more common in the mouse. We also show that there are differences in peptide expression in parasympathetic pelvic neurons between strains of mice. These studies provide the framework for future investigations on neural regulation of urogenital function, particularly in transgenic and knockout models.
Publisher: Media Peternakan
Date: 08-2018
Publisher: ZappyLab, Inc.
Date: 14-01-2019
DOI: 10.17504/PROTOCOLS.IO.W3DFGI6
Abstract: This protocol is used to visualise sensory and autonomic neurons innervating the bladder body (dome), bladder trigone or proximal urethra in an experimental adult male or female rat. The protocol is performed under anesthesia and should incorporate all local requirements for standards of animal experimentation, including methods of anesthesia, surgical environment, and post-operative monitoring and care.
Publisher: Frontiers Media SA
Date: 09-2015
Publisher: ZappyLab, Inc.
Date: 22-05-2020
DOI: 10.17504/PROTOCOLS.IO.BGRMJV46
Abstract: This collection describes the procedures required to implant a pelvic nerve array and bladder catheter into male Sprague-Dawley rats as well as cystometry and electrophysiological testing in awake animals. This collection includes protocols for: STAGE 1: Implantation of a pelvic nerve array in rats STAGE 2: Cystometry in awake rats STAGE 3: Electrophysiological recording of electrically-evoked compound action potentials
Publisher: IOP Publishing
Date: 15-05-2023
Abstract: Objective. Endovascular neuromodulation has attracted substantial interest in recent years as a minimally invasive approach to treat neurological disorders. In this study, we investigated with a computational model the feasibility of stimulating peripheral nerves with an endovascular stent-mounted electrode array. Approach. Anatomically realistic FEM models were constructed for the pudendal and vagal neurovascular bundles. The electromagnetic fields generated from electrical stimuli were computed using Sim4Life NEURON models to predict dynamic axonal responses. Main results. The models predict that the stimulation thresholds of the endovascular stent-electrode array configurations tested are comparable to that of ring electrodes and are dependent on the inter-electrode distance and orientation of the device. Arranging multiple electrodes along the longitudinal axis of the nerve lowers surface charge density without sacrificing axon recruitment, whereas arranging electrodes along the circumference of the blood vessel reduces the risk of misalignment but lowers axon recruitment. Significance. Overall, this study predicts that the endovascular stent-electrode array is a feasible stimulation option for peripheral nerves, and the electrode array can be flexibly optimized to achieve the lowest stimulation threshold.
Publisher: Springer Science and Business Media LLC
Date: 24-10-2017
Publisher: Wiley
Date: 11-06-2021
DOI: 10.1002/CNE.24949
Abstract: Storage and voiding of urine from the lower urinary tract (LUT) must be timed precisely to occur in appropriate behavioral contexts. A major part of the CNS circuit that coordinates this activity is found in the lumbosacral spinal cord. Immediate early gene (IEG) activity mapping has been widely used to investigate the lumbosacral LUT‐related circuit, but most reports focus on the effects of noxious stimulation in anesthetized female rats. Here we use c‐Fos and EGR‐1 (Zif268) activity mapping of lumbosacral spinal cord to investigate cystometry‐induced micturition in awake female and male rats. In females, after cystometry c‐Fos neurons in spinal cord segments L5–S2 were concentrated in the sacral parasympathetic nucleus (SPN), dorsal horn laminae II–IV, and dorsal commissural nucleus (SDCom). Comparisons of cystometry and control groups in male and female revealed sex differences. Activity mapping suggested dorsal horn laminae II–IV was activated in females but showed net inhibition in males. However, inhibition in male rats was not detected by EGR‐1 activity mapping, which showed low coexpression with c‐Fos. A class of catecholamine neurons in SPN and SDCom neurons were also more strongly activated by micturition in females. In both sexes, most c‐Fos neurons were identified as excitatory by their absence of Pax2 expression. In conclusion, IEG mapping in awake male and female rats has extended our understanding of the functional molecular anatomy of the LUT‐related circuit in spinal cord. Using this approach, we have identified sex differences that were not detected by previous studies in anesthetized rats.
Publisher: Elsevier BV
Date: 2018
Publisher: Hindawi Limited
Date: 23-02-2018
DOI: 10.1111/JAI.13650
Publisher: Ovid Technologies (Wolters Kluwer Health)
Date: 08-2013
Publisher: Elsevier BV
Date: 02-1994
Abstract: Adhesion to proteins of the extracellular matrix exerts a profound influence upon cell function and behavior. Similar adhesive interactions mediate the spreading of cultured cells upon artificial substrata. Recently we observed that thyrotropin (TSH) and intercellular contact regulated thyroid cell-substrate adhesion to inhibit cell spreading, but not initial attachment. This is a mechanism which preserves thyroid follicular differentiation in culture. In the present study we have investigated the role of cytoplasmic components in mediating thyroid cell adhesion to collagen. The earliest change associated with cell spreading was the accumulation of vinculin and phosphotyrosine in developing focal adhesions, which was followed by stress fiber and microtubule assembly. Genistein, an inhibitor of tyrosine kinases, and cytochalasin B inhibited cell spreading and focal adhesion formation without affecting initial attachment to substrate. In contrast microtubule disorganization by colchicine did not alter any parameter of thyroid cell-substrate adhesion. These observations indicate that protein tyrosine phosphorylation and dynamic microfilament integrity are essential for attached thyroid cells to spread upon substrate. They are therefore potential intracellular loci at which TSH and intercellular contact may regulate cell adhesion to extracellular matrix and influence thyroid cell behavior.
Publisher: IEEE
Date: 07-2020
Publisher: ZappyLab, Inc.
Date: 15-12-2019
DOI: 10.17504/PROTOCOLS.IO.BAHZIB76
Abstract: This protocol is suitable for preserving tissues for anatomical studies of organs, ganglia, spinal cord or brain in adult rats. The protocol is performed under anesthesia and should incorporate all local requirements for standards of animal experimentation.
Publisher: ZappyLab, Inc.
Date: 10-2021
DOI: 10.17504/PROTOCOLS.IO.BYQCPVSW
Abstract: This protocol is used for immunohistochemical visualisation of cholera toxin subunit B within afferents innervating the lower urinary tract in cryosections of rat lumbosacral spinal cord. Free-floating sections are processed in a double labelling protocol to distinguish regions of innervation by these afferents. Cholera toxin B antibody [lower urinary tract afferents] Choline acetyltransferase antibody [preganglionic autonomic neurons and motoneurons]
Publisher: Elsevier BV
Date: 10-2006
DOI: 10.1016/J.NEUROSCIENCE.2006.06.031
Abstract: Developmentally, semaphorin 3A (sema3A) is an important chemorepellent that guides centrally projecting axons of dorsal root ganglion (DRG) neurons. Sema3A-mediated growth cone collapse can be prevented by cyclic GMP (cGMP) and nerve growth factor (NGF) in embryonic neurons. Sema3A may also play a role in directing regrowth of injured axons in adults, and interactions with neurotrophic factors near the injury site may determine the extent and targeting of both regenerative and aberrant growth. The aim of this study was to determine whether NGF, glial cell line-derived neurotrophic factor (GDNF) and neurturin (NTN) modulate sema3A-mediated growth cone collapse in cultured adult rat DRG neurons. Sema3A caused a significant increase in growth cone collapse, which was completely prevented by prior treatment with NGF, GDNF or NTN. Immunocytochemical experiments showed that sema3A-sensitive neurons were heterogeneous in their expression of neurotrophic factor receptors and responses to neurotrophic factors, raising the possibility of novel, convergent signaling mechanisms between these substances. Increasing cGMP levels caused growth cone collapse, whereas sema3A-mediated collapse was prevented by inhibition of guanylate cyclase or by increasing cyclic AMP levels. In conclusion, sema3A signaling pathways in adult neurons differ to those described in embryonic neurons. Three different neurotrophic factors each completely prevent sema3A-mediated collapse, raising the possibility of novel converging signaling pathways. These studies also show that there is considerable potential for neurotrophic factors to regulate sema3A actions in the adult nervous system. This may provide insights into the mechanisms underling misdirected growth and targeting of sensory fibers within the spinal cord after injury, that is thought to contribute to development of autonomic dysreflexia and neuropathic pain.
Publisher: Elsevier BV
Date: 2019
Publisher: ZappyLab, Inc.
Date: 17-12-2019
DOI: 10.17504/PROTOCOLS.IO.BAKIICUE
Abstract: This collection describes the procedures required to visualise and characterize synaptic boutons associated with functionally classified pelvic ganglion autonomic neurons of adult male and female Sprague-Dawley rats. This collection includes protocols for: STAGE 1: Surgery to micro-inject fluorescent retrograde tracer dyes into one or more sites within the lower urinary tract STAGE 2: Intracardiac perfusion with fixative to preserve neural tissues of interest STAGE 3: Immunohistochemical labeling of thick cryosections of pelvic ganglia STAGE 4: Confocal microscopy and image analysis of synaptic boutons associated with ganglion neurons.
Publisher: African Journals Online (AJOL)
Date: 1970
Abstract: The objective of this study was to determine the effect of pomegranate (Punica granatum L.) seed oil (PSO) on gas and methane (CH4) production, ruminal fermentation and microbial populations under in vitro conditions. Three treatments consisting of a control diet containing 10 mg tallow (CON) the control diet with 5 mg PSO + 5 mg tallow (MPSO) and the control diet containing 10 mg PSO (HPSO) were compared. Ten mg of the experimental fat/oil s les were inserted into a gas-tight 100 mL plastic syringe containing 30 mL of an incubation inoculum and 250 mg of a basic substrate of a hay/concentrate (1/1, w/w) mixture. In vitro gas production was recorded over 0, 2, 4, 6, 8, 10, 12 and 24 h of incubation. After 24 hours, incubation was stopped, and methane production, pH, volatile fatty acids (VFAs) and microbial counts were measured in the inoculant. Gas production at 4, 6, 8, 10, 12 and 24 h incubation, metabolizable energy and in vitro organic matter disappearance increased linearly and quadratically as level of PSO increased. Furthermore, the 10 mg PSO (HPSO) decreased CH4 production by 21.0% compared with the control (CON) group. There were no significant differences in total and in idual VFA concentrations between different levels of PSO, except for butyric acid. After 24 h of incubation, methanogenesis decreased in the HPSO compared with the MPSO and CON treatments. In addition, total bacteria and protozoa counts increased with rising PSO levels, while population methanogenesis declined significantly. These results suggested that PSO could reduce methane emissions, which might be beneficial to nutrient utilization and growth in ruminants.
Publisher: Elsevier BV
Date: 2004
Publisher: Wiley
Date: 08-2005
DOI: 10.1111/J.1460-9568.2005.04260.X
Abstract: Neurturin (NTN) is an important neurotrophic factor for parasympathetic neurons however, no studies to date have investigated the signalling mechanisms downstream of GFRalpha2 and Ret activation underlying this neurotrophic support. This is particularly important for pelvic parasympathetic neurons, which are prone to injury during surgical procedures such as prostatectomy, and where there are no current therapies for axonal regeneration. To address this issue we have cultured dissociated adult rat pelvic ganglion neurons and also examined the structural changes in pelvic ganglion neurons after axotomy. Axotomised penile neurons deprived of target-derived support had smaller somata than intact neurons. Studies of cultured adult pelvic ganglion neurons also demonstrated that NTN stimulated soma growth. Further experiments showed that NTN reduced the up-regulation of tyrosine hydroxylase expression in cultured pelvic parasympathetic neurons. NTN stimulated the extension of neurites in cultured parasympathetic, but not sympathetic, pelvic ganglion neurons. Inhibition of phosphatidylinositol 3-kinase prevented initiation of neurite outgrowth, whereas inhibition of the mitogen-activated protein kinase and the Src family kinase pathways disrupted NTN-stimulated microtubule assembly. Surprisingly, NTN did not activate the transcription factor cAMP-response element binding protein (CREB), which is typically involved in neurotrophic signalling in sympathetic neurons. This is the first study to identify signalling pathways activated by NTN in adult parasympathetic neurons. Our results may lead to a better understanding of regenerative mechanisms in parasympathetic neurons, especially for those innervating urogenital organs. Our results also indicate that neurotrophic signalling in parasympathetic neurons is different from that in other types of peripheral neurons.
Publisher: Wiley
Date: 15-06-1985
Abstract: The distribution of mucosal nerve fibres containing vasoactive intestinal peptide (VIP), substance P, somatostatin, neuropeptide Y (NPY), and enkephalinlike immunoreactivity was mapped by conventional immunohistochemical techniques throughout the mucosa of the esophagus, stomach, small and large intestines, and gall bladder. In addition, the distributions of endocrine cells immunoreactive for three peptides localized by these antisera (namely somatostatin, pancreatic polypeptide, and substance P) were recorded. Tissues from guinea pigs, rats, dogs, marmosets, and humans were studied. It was hoped that this information would enable possible target tissues and functional roles for the peptides to be identified. In the mucosa, peptide nerve fibres were found throughout the lamina propria, including some which were close to the epithelium and others associated with small blood vessels. Although there was a general similarity of peptide nerve distribution between regions and species, many small variations were observed. VIP and substance P fibres were the most prevalent nerve type NPY fibres were also usually quite common. The distribution of somatostatin fibres was extremely variable between regions and species, and enkephalin fibres were usually rare. Endocrine cells of open (flask- or pyramid-shaped) and closed (rounded) types were seen basal cytoplasmic processes (of variable length) were seen on many cells immunoreactive for somatostatin or pancreatic polypeptide. Epithelial cells immunoreactive for substance P were seen in the dog, marmoset, and human. The distributions and shapes of endocrine cells varied widely between areas and species. These studies provide a basis for the correlation of nerve distribution with pharmacological and physiological studies.
Publisher: IEEE
Date: 11-2021
Publisher: Public Library of Science (PLoS)
Date: 10-08-2011
Publisher: The Company of Biologists
Date: 15-05-2015
DOI: 10.1242/DEV.117903
Abstract: Malformation of the urogenital tract represents a considerable paediatric burden, with many defects affecting the lower urinary tract (LUT), genital tubercle and associated structures. Understanding the molecular basis of such defects frequently draws on murine models. However, human anatomical terms do not always superimpose on the mouse, and the lack of accurate and standardised nomenclature is h ering the utility of such animal models. We previously developed an anatomical ontology for the murine urogenital system. Here, we present a comprehensive update of this ontology pertaining to mouse LUT, genital tubercle and associated reproductive structures (E10.5 to adult). Ontology changes were based on recently published insights into the cellular and gross anatomy of these structures, and on new analyses of epithelial cell types present in the pelvic urethra and regions of the bladder. Ontology changes include new structures, tissue layers and cell types within the LUT, external genitalia and lower reproductive structures. Representative illustrations, detailed text descriptions and molecular markers that selectively label muscle, nerves/ganglia and epithelia of the lower urogenital system are also presented. The revised ontology will be an important tool for researchers studying urogenital development/malformation in mouse models and will improve our capacity to appropriately interpret these with respect to the human situation.
Publisher: ZappyLab, Inc.
Date: 14-12-2021
DOI: 10.17504/PROTOCOLS.IO.B2UEQETE
Abstract: This collection includes the protocols required to map the lower urinary tract afferent projections to the lumbosacral spinal cord of male and female Sprague-Dawley rats. Afferents can be visualized using 3D reconstruction of alternating sections in TissueMaker (MBF Bioscience), or through the immunolabelling and clearing method, iDISCO. The following protocols are performed, regardless of endpoint: STAGE 1: Use of cholera toxin subunit B to label neural projections to lower urinary tract organs. STAGE 2: Intracardiac perfusion with fixative for anatomical studies. The next set of protocols pertain to the 3D reconstruction of spinal cord from alternating sections. STAGE 3: Immunohistochemical labelling of lower urinary tract afferents in spinal cord. STAGE 4: Quantitation of lower urinary tract afferents in 3D reconstruction of lumbosacral spinal cord sections For the visualization of lower urinary tract afferents in the intact spinal cord, skip Stages 3 and 4, and instead use Stage 5: STAGE 5: Immunolabelling and clearing of intact spinal cord for visualization of lower urinary tract afferents
Publisher: No publisher found
Date: 1996
Publisher: ZappyLab, Inc.
Date: 10-2021
DOI: 10.17504/PROTOCOLS.IO.BYQDPVS6
Abstract: The whole-mount immunolabeling and clearing method (iDISCO) was used to visualize cholera toxin subunit B-labelled lower urinary tract afferents in the lumbosacral spinal cord of the rat. Imaging of spinal cord was performed on a light sheet microscope with a 12x lens. Concurrently, choline acetyltransferase identified preganglionic autonomic neurons and motoneurons within the spinal cord, which were used to confirm the rostrocaudal location of afferents.
Publisher: Hindawi Limited
Date: 2014
DOI: 10.1155/2014/592709
Abstract: Two experiments were simultaneously conducted with Morus alba (white mulberry) foliage extract (MFE) as a growth promoter and treatment of Aeromonas hydrophila infection in separate 60 and 30 days trail (Experiments 1 and 2, resp.) in African catfish ( Clarias gariepinus ). In Experiment 1, four diets, control and control supplemented with 2, 5, or 7 g MFE/kg dry matter (DM) of diet, were used. In Experiment 2, fish were intraperitoneally infected with Aeromonas hydrophila and fed the same diets as experiment 1 plus additional two diets with or without antibiotic. Results of experiment 1 showed that growth was unaffected by dietary levels of MFE. Treatments with the inclusion of MFE at the levels of 5 and 7 g/Kg DM had no mortality. Red blood cells (RBC), albumin, and total protein were all higher for the treatments fed MFE (5 and 7 g/Kg DM). Results of experiment 2 showed RBC, hemoglobin, hematocrit, globulin, albumin, and total protein improved with the increase in MFE in the infected fish. The dietary MFE at the level of 7 g/kg DM reduced mortality rate. In conclusion, MFE at the level of 7 g/kg DM could be a valuable dietary supplement to cure the infected fish.
Publisher: Frontiers Media SA
Date: 24-10-2018
Publisher: Wiley
Date: 24-07-2014
DOI: 10.1002/CNE.23648
Publisher: Wiley
Date: 23-02-2009
DOI: 10.1002/CNE.21986
Abstract: Spinal cord injury commonly causes chronic, neuropathic pain. The mechanisms are poorly understood but may include structural plasticity within spinal and supraspinal circuits. Our aim was to determine whether structural remodeling within the dorsal horn rostral to an incomplete injury differs from a complete spinal cord transection. Four immunohistochemical populations of primary afferent C-fibers, and descending catecholamine and serotonergic projections, were examined in segments T9-T12 at 2 and 12 weeks after a T13 clip-compression injury in adult male rats. Dorsal root ganglia were also examined. Two weeks after injury, fibers immunoreactive for calcitonin gene-related peptide (CGRP) or GDNF-family receptors (GFRalpha1, GFRalpha2, GFRalpha3) showed distinct injury responses within the superficial dorsal horn. CGRP fibers decreased, but GFRalpha1, GFRalpha2 and GFRalpha3 fibers did not change. In contrast, all groups were decreased by 12 weeks after injury. Catecholamine fibers showed a decrease at 2 weeks followed by an increase in density at 12 weeks, whereas serotonergic fibers showed a decrease (restricted to deep dorsal horn) at 12 weeks. These results show that the dorsal horn of the spinal cord undergoes substantial structural plasticity rostral to a compression injury, with the most profound effect being a prolonged and possibly permanent loss of primary afferent fibers. This loss was more extensive and more prolonged than the loss that follows spinal cord transection. Our results provide further evidence that anatomical reorganization of sensory and nociceptive dorsal horn circuits rostral to an injury could factor in the development or maintenance of spinal cord injury pain.
Publisher: Elsevier BV
Date: 04-2015
DOI: 10.1016/J.EXPNEUROL.2015.02.026
Abstract: Following peripheral nerve injury, restoration of function may occur via the regeneration of injured axons or compensatory sprouting of spared axons. Injury to visceral nerves that control urogenital organs is a common consequence of pelvic surgery, however their capacity to reinnervate organs is poorly understood. To determine if and how sensory and motor connections to the bladder are re-established, a novel surgical model of visceral nerve injury was performed unilaterally in adult male Wistar rats. Bladder-projecting motor and sensory neurons in pelvic ganglia and lumbosacral dorsal root ganglia, respectively, were identified and characterised by retrograde tracing and immunofluorescence. Application of tracers ipsi- and contralateral to injury distinguished the projection pathways of new connections in the bladder. In naive animals, the majority of sensory and motor neurons project ipsilaterally to the bladder, while ~20 % project contralaterally and ~5 % bilaterally. Injured axons of motor neurons were unable to regenerate by 4weeks after transection. In contrast, by this time many injured sensory neurons regrew axons to reform a substantial plexus within the detrusor and suburothelial tissues. These regeneration responses were also indicated by upregulation of activating transcription factor-3 (ATF-3), which was sustained in motor neurons but transient in sensory bladder-projecting neurons. Axotomy had little or no effect on the survival of bladder-projecting sensory and motor neurons. We also found evidence that uninjured motor and sensory neurons develop additional projections to the denervated bladder tissue and return connectivity, likely by undergoing compensatory growth. In conclusion, our results show that visceral sensory and motor neurons have a different capacity to regenerate axons following axotomy, however in both components of the circuit uninjured bladder neurons spontaneously grow new axon collaterals to replace the lost terminal field within the organ. For a full functional recovery, understanding the environmental and cellular mechanisms that reduce the ability of pelvic ganglion cells to undergo axonal regeneration is needed.
Publisher: Wiley
Date: 2008
DOI: 10.1002/CNE.21593
Abstract: We have investigated the development of autonomic nerves in the urogenital tract of male mice and the effect of neurturin gene deletion on this process. At birth, autonomic innervation of the reproductive organs was sparse, but urinary bladder smooth muscle was well innervated. Further innervation of reproductive tissues occurred until P21, but noradrenergic axons established their complete terminal field later than nitrergic cholinergic axons: in adults the former are more prevalent, yet this became apparent only at P7 (vas deferens, seminal vesicles), P14 (prostate) or after P14 (penis). Neurturin was essential for initial projection of axons (mucosa of vas deferens), maintenance of terminal fields (prostate and seminal vesicles), or both functions (cavernosum of penis). In contrast, some targets (e.g., bladder muscle and suburothelium, vas deferens smooth muscle) were unaffected by neurturin gene deletion. Pelvic ganglion neurons more than doubled between birth and adulthood, probably as aresult of continued maturation of p75-positive undifferentiated neuronal precursors rather than cell ision. The adult number of neurons was achieved by P7 (sympathetic) or P21 (parasympathetic). In adult neurturin knockout mice, there were approximately 25% fewer parasympathetic neurons compared with wild types, because of failure of differentiation after P14. This study revealed the complexity of postnatal maturation of urogenital innervation, with each organ showing a distinct chronology of innervation and different requirement for neurturin. Our results also indicate that in adults there will be distinct differences in neurturin dependence between organs, such that proregenerative therapies may have to be tailored specifically for the nerve pathway of interest.
Publisher: Elsevier BV
Date: 03-2010
DOI: 10.1016/J.NEUROSCIENCE.2009.12.019
Abstract: A conditioning lesion improves regeneration of central and peripheral axons of dorsal root ganglion (DRG) neurons after a subsequent injury by enhancing intrinsic growth capacity. This enhanced growth state is also observed in cultured DRG neurons, which support a more sparsely and rapidly elongating mode of growth after a prior conditioning lesion in vivo. Here we examined differences in the capacity or requirements of specific types of sensory neurons for regenerative growth, which has important consequences for development of strategies to improve recovery after injury. We showed that after partial or complete injury of the sciatic nerve in mice, an elongating mode of growth in vitro was activated only in DRG neurons that did not express calcitonin gene-related peptide (CGRP) or bind Bandeiraea simplicifolia I-isolectin B4 (IB4). We also directly examined the response of conditioned sensory neurons to nerve growth factor (NGF), which does not enhance growth in injured peripheral nerves in vivo. We showed that after partial injury, NGF stimulated a highly branched and linearly restricted rather than elongating mode of growth. After complete injury, the function of NGF was impaired, which immunohistochemical studies of DRG indicated was at least partly due to downregulation of the NGF receptor, tropomyosin-related kinase A (TrkA). These results suggest that, regardless of the type of conditioning lesion, each type of DRG neuron has a distinct intrinsic capacity or requirement for the activation of rapidly elongating growth, which does not appear to be influenced by NGF.
Publisher: Wiley
Date: 23-05-2007
Publisher: Frontiers Media SA
Date: 09-03-2023
DOI: 10.3389/FNINS.2023.1072779
Abstract: A thorough understanding of the neuroanatomy of peripheral nerves is required for a better insight into their function and the development of neuromodulation tools and strategies. In biophysical modeling, it is commonly assumed that the complex spatial arrangement of myelinated and unmyelinated axons in peripheral nerves is random, however, in reality the axonal organization is inhomogeneous and anisotropic. Present quantitative neuroanatomy methods analyze peripheral nerves in terms of the number of axons and the morphometric characteristics of the axons, such as area and diameter. In this study, we employed spatial statistics and point process models to describe the spatial arrangement of axons and Sinkhorn distances to compute the similarities between these arrangements (in terms of first- and second-order statistics) in various vagus and pelvic nerve cross-sections. We utilized high-resolution transmission electron microscopy (TEM) images that have been segmented using a custom-built high-throughput deep learning system based on a highly modified U-Net architecture. Our findings show a novel and innovative approach to quantifying similarities between spatial point patterns using metrics derived from the solution to the optimal transport problem. We also present a generalizable pipeline for quantitative analysis of peripheral nerve architecture. Our data demonstrate differences between male- and female-originating s les and similarities between the pelvic and abdominal vagus nerves.
Publisher: Informa UK Limited
Date: 02-04-2016
Publisher: MyJove Corporation
Date: 07-03-2020
DOI: 10.3791/60904
Publisher: Hindawi Limited
Date: 18-04-2017
DOI: 10.1111/ARE.13266
Publisher: ZappyLab, Inc.
Date: 17-12-2019
DOI: 10.17504/PROTOCOLS.IO.BAKDICS6
Abstract: This protocol describes confocal microscopy and image analysis procedures for characterizing neuronal cell bodies and their associated synaptic boutons in thick (50 µm) cryosections. The protocol has been applied to rat pelvic ganglia, where neuronal cell bodies have been identified using immunohistochemical markers of specific neuron populations and/or fluorescent retrograde tracer.
Publisher: IOP Publishing
Date: 25-11-2021
Abstract: Objective. Neuromodulation of visceral nerves is being intensively studied for treating a wide range of conditions, but effective translation requires increasing the efficacy and predictability of neural interface performance. Here we use computational models of rat visceral nerve to predict how neuroanatomical variability could affect both electrical stimulation and recording with an experimental planar neural interface. Approach. We developed a hybrid computational pipeline, Vi sceral N erve E nsemble R ecording and S timulation (ViNERS), to couple finite-element modelling of extracellular electrical fields with biophysical simulations of in idual axons. Anatomical properties of fascicles and axons in rat pelvic and vagus nerves were measured or obtained from public datasets. To validate ViNERS, we simulated pelvic nerve stimulation and recording with an experimental four-electrode planar array. Main results. Axon diameters measured from pelvic nerve were used to model a population of myelinated and unmyelinated axons and simulate recordings of electrically evoked single-unit field potentials (SUFPs). Across visceral nerve fascicles of increasing size, our simulations predicted an increase in stimulation threshold and a decrease in SUFP litude. Simulated threshold changes were dominated by changes in perineurium thickness, which correlates with fascicle diameter. We also demonstrated that ViNERS could simulate recordings of electrically-evoked compound action potentials (ECAPs) that were qualitatively similar to pelvic nerve recording made with the array used for simulation. Significance. We introduce ViNERS as a new open-source computational tool for modelling large-scale stimulation and recording from visceral nerves. ViNERS predicts how neuroanatomical variation in rat pelvic nerve affects stimulation and recording with an experimental planar electrode array. We show ViNERS can simulate ECAPS that capture features of our recordings, but our results suggest the underlying NEURON models need to be further refined and specifically adapted to accurately simulate visceral nerve axons.
Publisher: Informa UK Limited
Date: 2014
Publisher: ZappyLab, Inc.
Date: 10-2021
DOI: 10.17504/PROTOCOLS.IO.BYQFPVTN
Abstract: This protocol details the 3D reconstruction of the lumbosacral spinal cord using alternating cryosections, and then goes through the steps required to quantify lower urinary tract afferents. Using TissueMaker (MBF Bioscience), images of alternating sections can be ordered and aligned prior to the production of a single image stack. In Neurolucida 360 (MBF Bioscience), regions of interest can be defined within the image stack, and the bouton-like immunolabelling of cholera toxin B can be segmented. Once saved, this data can then be extracted using Neurolucida Explorer (MBF Bioscience).
Publisher: Elsevier BV
Date: 2005
DOI: 10.1016/J.NEUROSCIENCE.2004.11.043
Abstract: DCC (deleted in colorectal cancer)-the receptor of the netrin-1 neuronal guidance factor-is expressed and is active in the central nervous system (CNS) during development, but is down-regulated during maturation. The substantia nigra contains the highest level of netrin-1 mRNA in the adult rodent brain, and corresponding mRNA for DCC has also been detected in this region but has not been localized to any particular neuron type. In this study, an antibody raised against DCC was used to determine if the protein was expressed by adult dopamine neurons, and identify their distribution and projections. Significant DCC-immunoreactivity was detected in midbrain, where it was localized to ventrally displaced A9 dopamine neurons in the substantia nigra, and ventromedial A10 dopamine neurons predominantly situated in and around the interfascicular nucleus. Strong immunoreactivity was not detected in dopamine neurons found elsewhere, or in non-dopamine-containing neurons in the midbrain. Terminal fields selectively labeled with DCC antibody corresponded to known nigrostriatal projections to the dorsolateral striatal patches and dorsomedial shell of the accumbens, and were also detected in prefrontal cortex, septum, lateral habenular and ventral pallidum. The unique distribution of DCC-immunoreactivity in adult ventral midbrain dopamine neurons suggests that netrin-1/DCC signaling could function in plasticity and remodeling previously identified in dopamine projection pathways. In particular, a recent report that DCC is regulated through the ubiquitin-proteosome system via Siah/Sina proteins, is consistent with a potential involvement in genetic and sporadic forms of Parkinson's disease.
Publisher: Elsevier BV
Date: 02-2017
Publisher: Springer Science and Business Media LLC
Date: 30-10-2015
Publisher: Elsevier BV
Date: 02-2017
Publisher: Frontiers Media SA
Date: 2017
Publisher: Springer Science and Business Media LLC
Date: 12-2009
Publisher: The Royal Society
Date: 29-07-1998
Abstract: The electroreceptors located in the bill skin of the platypus are modified secretory glands. The electroreceptive nerve terminals form bare endings in close proximity to the duct of these glands. In this study, we describe the autonomic innervation of the glands and a separate specialized autonomic innervation of the epidermal portion of the glandular duct. A range of immunohistochemical labels showed that the gland cells of the electroreceptors have a non–noradrenergic (putative parasympathetic) innervation. Phalloidin labelling revealed a ‘phincter’ of epidermal luminal cells that labelled strongly for actin. These actin–dense keratinocytes were seen to have a noradrenergic (putative sympathetic) innervation. Fine–diameter sensory fibres containing substance P (presumably C–fibre thermoreceptors or polymodal nociceptors) were observed to terminate in the superficial epidermis surrounding the pore of the gland. When the bill of the platypus is dry these pores were closed. However, when room temperature water was washed over the bill, the pores opened. It is proposed that this autonomic and sensory innervation, along with the actin sphincter, mediates the opening and closing of the pores. By doing this, the platypus prevents the desiccation of the bare electrosensory nerve terminals when it is out of the water, and it may also be a way to regulate the impedance of the internal electrical circuit presented to the water at the pores.
Publisher: ZappyLab, Inc.
Date: 17-08-2020
DOI: 10.17504/PROTOCOLS.IO.BJVXKN7N
Abstract: A pelvic nerve array is custom designed for implantation onto the pelvic nerve of male rats. This device is used for long-term (recovery) experiments. The device can electrically stimulate the pelvic nerve as well as record electrically evoked or spontaneous neural activity. Previously, we have successfully implanted rats for 2 months with no reports of biological adverse complications or damage sustained to the array. The surgical procedure is performed under anesthesia and should incorporate all local requirements for standards of animal experimentation, including methods of anesthesia, surgical environment, and post-operative monitoring and care.
Publisher: Oxford University Press (OUP)
Date: 12-2017
DOI: 10.1016/J.JSXM.2017.10.071
Abstract: Controversy exists in the literature regarding the presence or absence of an anatomic “G-spot.” However, few studies have examined the detailed topographic or histologic anatomy of the putative G-spot location. To determine the anatomy of the anterior vaginal wall and present detailed, systematic, accessible findings from female cadaveric dissections to provide anatomic clarity with respect to this location. Systematic anatomic dissections were performed on 13 female cadavers (32–97 years old, 8 fixed and 5 fresh) to characterize the gross anatomy of the anterior vaginal wall. Digital photography was used to document dissections. Dissection preserved the anterior vaginal wall, urethra, and clitoris. In 9 cadavers, the vaginal epithelial layer was reflected to expose the underlying urethral wall and associated tissues. In 4 cadavers, the vaginal wall was left intact before preservation. Once photographed, 8 specimens were transversely sectioned for macroscopic inspection and histologic examination. The presence or absence of a macroscopic anatomic structure at detailed cadaveric pelvis dissection that corresponds to the previously described G-spot and gross anatomic description of the anterior vaginal wall. Deep to the lining epithelium of the anterior vaginal wall is the urethra. There is no macroscopic structure other than the urethra and vaginal wall lining in the location of the putative G-spot. Specifically, there is no apparent erectile or “spongy” tissue in the anterior vaginal wall, except where the urethra abuts the clitoris distally. The absence of an anatomic structure corresponding to the putative G-spot helps clarify the controversy on this subject. Limitations to this study include limited access to specimens immediately after death and potential for observational bias. In addition, age, medical history, and cause of death are not publishable for privacy reasons. However, it is one of the most thorough and complete anatomic evaluations documenting the anatomic detail of the anterior vaginal wall. The G-spot, in its current description, is not identified as a discrete anatomic entity at macroscopic dissection of the urethra or vaginal wall. Further insights could be provided by histologic study.
Publisher: Elsevier BV
Date: 2018
DOI: 10.1016/J.FOODCHEM.2017.07.061
Abstract: A 9-week study was conducted to compare dietary corn starch (CS) or tapioca starch (TS), with or without being pre-gelatinized (PG), on the growth, feeding efficiencies, plasma and muscle biochemistry, intestinal short chain fatty acids (SCFA), and liver glycogen of triplicate groups of 20 red hybrid tilapia (Orecohromis sp.). Various pellet characteristics were evaluated, along with their surface and cross sectional microstructure. The PG diets had significantly higher water stability, bulk density, and protein solubility, along with a smoother surface. Tilapia fed the TS diet had lower growth than had all other tilapia, but were significantly improved when diet was pre-gelatinized. In the PG treatments, intestinal SCFA significantly decreased while plasma glucose, cholesterol and triglycerides, as well as liver glycogen, significantly increased. Fish fed the CS diet had significantly more long chain polyunsaturated fatty acid than had those fed by other treatments. Pre-gelatinization may improve fish productivity and offer greater flexibility during aquafeed production.
Publisher: ZappyLab, Inc.
Date: 17-12-2019
DOI: 10.17504/PROTOCOLS.IO.BAKXICXN
Abstract: This collection describes the procedures required to visualize and characterize lumbosacral spinal neurons that are activated by cystometry of awake adult male and female Sprague-Dawley rats. This collection includes protocols for: STAGE 1: Surgery to cannulate the bladder, followed by recovery then cystometry STAGE 2: Intracardiac perfusion with fixative to preserve the spinal cord tissue STAGE 3: Immunohistochemical labelling of spinal cord sections to visualise immediate early gene expression in specific spinal regions and neuronal populations STAGE 4: Microscopy and image analysis to assess patterns of immediate early gene expression in different spinal cord regions
Publisher: Hindawi Limited
Date: 2015
DOI: 10.1155/2015/897639
Abstract: Probiotics are live microorganisms that exert beneficial effects on the host, when administered in adequate amounts. Mostly, probiotics affect the gastrointestinal (GI) tract of the host and alter the composition of gut microbiota. Nowadays, the incidence of hip fractures due to osteoporosis is increasing worldwide. Ovariectomized (OVX) rats have fragile bone due to estrogen deficiency and mimic the menopausal conditions in women. Therefore, this study aimed to examine the effects of Bifidobacterium longum ( B. longum ) on bone mass density (BMD), bone mineral content (BMC), bone remodeling, bone structure, and gene expression in OVX rats. The rats were randomly assigned into 3 groups (sham, OVX, and the OVX group supplemented with 1 mL of B. longum 10 8 –10 9 colony forming units (CFU)/mL). B. longum was given once daily for 16 weeks, starting from 2 weeks after the surgery. The B. longum supplementation increased ( p 0.05 ) serum osteocalcin (OC) and osteoblasts, bone formation parameters, and decreased serum C-terminal telopeptide (CTX) and osteoclasts, bone resorption parameters. It also altered the microstructure of the femur. Consequently, it increased BMD by increasing ( p 0.05 ) the expression of Sparc and Bmp-2 genes. B. longum alleviated bone loss in OVX rats and enhanced BMD by decreasing bone resorption and increasing bone formation.
Publisher: Elsevier BV
Date: 2018
Publisher: Elsevier BV
Date: 12-2017
Publisher: Hindawi Limited
Date: 28-07-2018
DOI: 10.1111/ANU.12806
Publisher: Elsevier
Date: 2018
Publisher: Society for Neuroscience
Date: 2020
DOI: 10.1523/ENEURO.0397-19.2019
Abstract: Visceral sensory neurons encode distinct sensations from healthy organs and initiate pain states that are resistant to common analgesics. Transcriptome analysis is transforming our understanding of sensory neuron subtypes but has generally focused on somatic sensory neurons or the total population of neurons in which visceral neurons form the minority. Our aim was to define transcripts specifically expressed by sacral visceral sensory neurons, as a step towards understanding the unique biology of these neurons and potentially leading to identification of new analgesic targets for pelvic visceral pain. Our strategy was to identify genes differentially expressed between sacral dorsal root ganglia (DRG) that include somatic neurons and sacral visceral neurons, and adjacent lumbar DRG that comprise exclusively of somatic sensory neurons. This was performed in adult and E18.5 male and female mice. By developing a method to restrict analyses to nociceptive Trpv1 neurons, a larger group of genes were detected as differentially expressed between spinal levels. We identified many novel genes that had not previously been associated with pelvic visceral sensation or nociception. Limited sex differences were detected across the transcriptome of sensory ganglia, but more were revealed in sacral levels and especially in Trpv1 nociceptive neurons. These data will facilitate development of new tools to modify mature and developing sensory neurons and nociceptive pathways.
Publisher: Springer Science and Business Media LLC
Date: 27-03-2017
DOI: 10.1007/S10695-017-0365-0
Abstract: Dietary organic acids are increasingly being investigated as a potential means of improving growth and nutrient utilization in aquatic animals. A 9-week study was performed to compare equal amounts (2%) of different organic acids (sodium butyrate, acetate, propionate, or formate) on the growth, muscle proximate composition, fatty acid composition, cholesterol and lipid peroxidation, differential cell counts, plasma biochemistry, intestinal short-chain fatty acid (SCFA) level, and liver histopathology to red hybrid tilapia (Oreochromis sp.) (initial mean weight of 2.87 g). A second experiment was performed to determine their effects on lipid peroxidation and trimethylamine (TMA) when added at 1% to tilapia meat and left out for 24 h. The results of the first experiment showed no treatment effect to growth, feeding efficiencies, or muscle fatty acid composition, but all dietary organic acids significantly decreased intestinal SCFA. Dietary butyrate and propionate significantly decreased muscle lipid peroxidation compared to the control group, but the dietary formate treatment had the lowest lipid peroxidation compared to all treatments. Muscle crude protein and lipid in tilapia fed the formate diet were significantly lower and higher, respectively, and showed evidence of stress based on the differential cell counts, significantly higher plasma glucose and liver glycogen, as well as inflammatory responses in the liver. Although a potential benefit of dietary organic acids was a reduction to lipid peroxidation, this could be accomplished post-harvest by direct additions to the meat. In addition, inclusions of butyrate and propionate to tilapia meat significantly decreased TMA, which might be a more cost-effective option to improve the shelf life of tilapia products.
Publisher: Elsevier BV
Date: 2004
DOI: 10.1016/J.NEUROSCIENCE.2004.08.019
Abstract: Actions of gonadal steroids have not been widely investigated in the peripheral nervous system, although many dorsal root ganglion (DRG) and autonomic pelvic ganglion (PG) neurons express estrogen receptors (ERs). We have studied the effects of 17beta-estradiol exposure on cultured DRG and PG neurons from adult rats. Western blotting analysis of DRG extracts detected phosphorylation of ERK1 and ERK2 (extracellular signal-regulated kinases) that peaked 10 min after exposure to 17beta-estradiol. These extracts contain both neurons and glia therefore, to determine if this response occurred in DRG neurons, we developed an immunocytochemical method to specifically measure activation in in idual neurons. These measurements showed that estradiol increased phosphorylation of CREB (cyclic AMP response-element binding protein), which was consistently blocked by the ERK pathway inhibitor PD98059 but not by the inhibitors of phosphatidylinositol 3-kinase, wortmannin and LY294002. 17beta-Estradiol activation of CREB in DRG neurons was reduced by the ER antagonist, ICI182780. In contrast, in PG neurons estradiol did not affect CREB phosphorylation, highlighting a difference in E2 responses in different populations of peripheral neurons. This study has shown that estrogens can rapidly activate signaling pathways associated with CREB-mediated transcriptional regulation in sensory neurons. As these pathways also mediate many effects of neurotrophic factors, changes in estrogen levels (e.g. during puberty, pregnancy or menopause) could have broad-ranging genomic and non-genomic actions on urogenital pain sensation and reflex pathways.
Publisher: Ovid Technologies (Wolters Kluwer Health)
Date: 10-2010
DOI: 10.1016/J.PAIN.2010.07.007
Abstract: Spinal cord injury (SCI) is a major cause of persistent neuropathic pain of central origin. Recent evidence suggests neuropathic pain in clinically complete SCI patients correlates with limited sensory function below the lesion (sensory discomplete). On this basis we examined if the onset of mechanical hyperalgesia was different in rodents after a severe incomplete clip-compression SCI versus a complete spinal cord transection at thoracic segment T13. Above-level withdrawal behaviors evoked by forepaw stimulation provided evidence of mechanical hyperalgesia after incomplete but not complete SCI, whereas below-level responses evoked by hindpaw stimulation revealed hypersensitivity after both injuries. The latency of the above-level response was 4-5 wks but was longer after a moderate clip-compression injury. Mechanical hyperalgesia was fully reversed by three analgesic drugs used in treating neuropathic SCI pain, but their duration of action differed significantly, showing a rank order of amitriptyline (24-48 h)≫morphine (6 h)>gabapentin (2 h). Evidence of central sensitization in cervical spinal cord segments that receive sensory projections from the forelimbs was provided by immunohistochemistry for Zif268, a functional marker of neuroplasticity. Zif268-immunoreactive neurons in laminae I/II increased in response to repetitive noxious forepaw stimulation in the incomplete SCI group, and this response was reduced in the complete transection and sham-operated groups. These data are consistent with the hypothesis that neuropathic pain of cord origin is more likely to develop after SCI when there is an incomplete loss of axons traversing the lesion.
Publisher: Wiley
Date: 04-10-2021
DOI: 10.1002/NAU.24807
Publisher: ZappyLab, Inc.
Date: 14-01-2019
DOI: 10.17504/PROTOCOLS.IO.W3EFGJE
Abstract: This protocol is used for studies of neurons containing retrograde tracer dye in adult rat dorsal root ganglia (DRG) or pelvic ganglia (PG synonym, major pelvic ganglia). Cryosections of ganglia are mounted directly on slides for immunohistochemical staining. Antibodies have been selected to distinguish different neurochemical classes of PG neurons or to specifically identify subclasses of sensory neurons that are myelinated (immunoreactive for NF200 [neurofilament, 200 kD]) or nociceptive (immunoreactive for TRPV1).
Publisher: ZappyLab, Inc.
Date: 16-12-2019
DOI: 10.17504/PROTOCOLS.IO.BAKCICSW
Abstract: This protocol describes immunohistochemical procedures applied to thick (50 µm) cryosections mounted directly on slides. It is used when the structures to be analysed are too large to remain intact within thin (10-20 µm) cryosections. Antibodies have been selected to distinguish different neurochemical classes of autonomic ganglion neurons and synaptic boutons associated with these neurons. The protocol can also be used to characterize neurons containing retrograde tracer.
Publisher: Frontiers Media SA
Date: 17-12-2020
DOI: 10.3389/FNINS.2020.619275
Abstract: Bioelectronic medical devices are well established and widely used in the treatment of urological dysfunction. Approved targets include the sacral S3 spinal root and posterior tibial nerve, but an alternate target is the group of pelvic splanchnic nerves, as these contain sacral visceral sensory and autonomic motor pathways that coordinate storage and voiding functions of the bladder. Here, we developed a device suitable for long-term use in an awake rat model to study electrical neuromodulation of the pelvic nerve (homolog of the human pelvic splanchnic nerves). In male Sprague-Dawley rats, custom planar four-electrode arrays were implanted over the distal end of the pelvic nerve, close to the major pelvic ganglion. Electrically evoked compound action potentials (ECAPs) were reliably detected under anesthesia and in chronically implanted, awake rats up to 8 weeks post-surgery. ECAP waveforms showed three peaks, with latencies that suggested electrical stimulation activated several subpopulations of myelinated A-fiber and unmyelinated C-fiber axons. Chronic implantation of the array did not impact on voiding evoked in awake rats by continuous cystometry, where void parameters were comparable to those published in naïve rats. Electrical stimulation with chronically implanted arrays also induced two classes of bladder pressure responses detected by continuous flow cystometry in awake rats: voiding contractions and non-voiding contractions. No evidence of tissue pathology produced by chronically implanted arrays was detected by immunohistochemical visualization of markers for neuronal injury or noxious spinal cord activation. These results demonstrate a rat pelvic nerve electrode array that can be used for preclinical development of closed loop neuromodulation devices targeting the pelvic nerve as a therapy for neuro-urological dysfunction.
Publisher: Elsevier BV
Date: 08-2019
DOI: 10.1016/J.MEATSCI.2019.04.008
Abstract: This study investigated the carcass characteristics, physico-chemical properties, storage stability and cholesterol content of meat from goats fed with different levels of naturally-produced lovastatin used to mitigate enteric methane production. Twenty intact Saanen male goats of 5-6 months old with initial live weight of 25.8 ± 4.0 kg were randomly allotted into four dietary treatments containing 0 (Control), 2 (Low), 4 (Medium) and 6 mg (High) per kg live weight (LW) of naturally-produced lovastatin for 12 consecutive weeks. No differences were found in all the parameters measured except for full LW, hot and cold carcass weight, shear force, color and cholesterol content among the treatment groups. Aging had significant effects on all the parameters measured in this study except a* (redness) of meat. Meat s les in the Medium and High treatments were of higher lightness and yellowness, more tender and lower cholesterol levels. We conclude that, in addition to mitigate enteric methane emissions, dietary supplementation of naturally-produced lovastatin at 4 mg/kg LW could be a feasible feeding strategy to produce tender meat containing lower cholesterol.
Publisher: Wiley
Date: 08-08-1993
Abstract: The distribution of neurons and endocrine cells containing various peptides or catecholamines was examined in the digestive tracts of the echidna (Tachyglossus aculeatus) and the platypus (Ornithorhynchus anatinus). Comparisons were made with published studies in other species in order to obtain a broader view of the phylogenetic distribution and possible functions of gut peptides and catecholamines. Further comparisons between the echidna and platypus were made in light of their different dietary features and gut histology. The distribution of neurons and axons containing catecholamines or various peptides resembled that in other species (such as the frequent appearance of axons containing substance P and vasoactive intestinal peptide in the intestinal mucosa, and axons containing substance P or enkephalins in the circular muscle). In both species, the stomach histologically resembles the esophagus, being aglandular and lined with stratified squamous epithelium. Innervation of these two organs was similar but not identical, with a greater array of peptides found in the gastric muscle. The intestinal mucosa was densely innervated in both species. The platypus small intestine is unusual in having a thick and deeply folded mucosa (but no villi), in which the superficial epithelium is absent or incomplete at many sites many axons travel close to these luminal surfaces. Many (putative noradrenergic) axons associated with blood vessels contained neuro-peptide Y, but there was no evidence for intrinsic catecholamine-containing neurons. Somatostatin and cholecystokinin were present in some endocrine cells, but unlike many mammals, absent in neuronal tissue. These studies have shown that there are many strong similarities between monotremes and other mammals in the distribution and array of peptides found within nervous and endocrine tissues of the digestive tract. However, numerous small differences of the echidna and platypus innervation may be correlated with their different digestive structures.
Publisher: Wiley
Date: 2005
DOI: 10.1002/CNE.20552
Abstract: Enkephalin (ENK)-immunoreactive (IR) axons occur in regions containing spinal autonomic neurons and endogenous opiates contribute to spinal regulation of bladder function. To identify possible spinal sites of opiate action, we used immunocytochemistry for ENK with retrograde tracing from the major pelvic ganglion (MPG), a key location for postganglionic neurons controlling pelvic viscera, with cholera toxin B subunit (CTB) or CTB-horseradish peroxidase (CTB-HRP). We compared the relationship of ENK-IR axons with sympathetic preganglionic neurons (SPNs) projecting to the MPG between intact spinal cords and cords with 2- or 11-week complete transections between thoracic segments 4 and 5. By light microscopy, sections of intact cord showed dense networks of ENK-IR axons surrounding CTB-IR SPNs in the intermediolateral cell column (IML), intercalated nucleus, and central autonomic area of lower thoracic and upper lumbar cord. This staining pattern was similar in rats with 2- or 11-week transections. Ultrastructurally, ENK-IR axons formed synapses on SPNs in all three autonomic subnuclei of intact cord. In the IML, ENK-IR varicosities contributed 52% of the synapses on the somata of MPG-projecting SPNs. In 2-week transected cord, synapses from ENK-IR axons persisted on SPNs and the proportion of input to IML SPNs had increased to 67%, probably reflecting loss of supraspinal input. These results suggest that endogenous opioids could play a major role in controlling sympathetic outflow to the bladder through a direct action on SPNs. The persistence of the dense ENK innervation after complete cord transection indicates that the ENK-IR input to SPNs arises predominantly from intraspinal sources.
Publisher: ZappyLab, Inc.
Date: 10-2021
DOI: 10.17504/PROTOCOLS.IO.BYQBPVSN
Abstract: This protocol is used to visualise sensory and autonomic neurons innervating organs of the lower urinary tract in an experimental adult male or female rat. The protocol is performed under anesthesia and should incorporate all local requirements for standards of animal experimentation, including methods of anesthesia, surgical environment, and post-operative monitoring and care.
Publisher: Elsevier
Date: 2006
Publisher: ZappyLab, Inc.
Date: 14-01-2019
DOI: 10.17504/PROTOCOLS.IO.W3FFGJN
Abstract: This protocol is suitable for preserving tissues for anatomical studies of organs, ganglia, spinal cord or brain in adult rats. The protocol is performed under anesthesia and should incorporate all local requirements for standards of animal experimentation.
Publisher: Wiley
Date: 22-05-1992
Abstract: Many visceral afferent neurons contain peptides, which have been proposed as histochemical markers for nerve pathways of particular targets or as transmitter candidates. The former possibility was investigated in the present study. Primary afferent neurons which project to the urinary bladder, distal colon or penis of rats, and the colon of cats were labelled with retrogradely transported fluorescent dyes (Fast Blue, True Blue, or Fluoro Gold). One to six weeks after dye injection into the organs, lumbosacral dorsal root ganglia were removed, treated with colchicine, and processed for immunohistochemical identification of five peptides. Dye-labelled neurons were distributed in an organ-specific manner in the lower lumbosacral ganglia, where colon afferent neurons were almost exclusively found in S1 ganglia, penis neurons primarily in L6, and bladder neurons at both levels. Substance P- (SP), calcitonin gene-related peptide-(CGRP), vasoactive intestinal peptide- (VIP), enkephalin- (ENK), and somatostatin- (SOM) immunoreactivity (IR) were detected in neurons in all lumbosacral ganglia but only some of these peptides were present in a large percentage of labelled neurons. The numbers of peptide-containing neurons innervating each organ were CGRP greater than SP greater than VIP greater than ENK greater than SOM however some differences were observed in the relative proportions of these neuronal populations between upper lumbar and lower lumbosacral ganglia and between different organs. The major difference seen at the upper lumbar level was amongst the SP-IR neurons, which were common (25-30%) amongst bladder and colon afferent neurons but absent in penis neurons.(ABSTRACT TRUNCATED AT 250 WORDS)
Publisher: The Endocrine Society
Date: 10-07-2008
DOI: 10.1210/EN.2008-0278
Abstract: There is mounting evidence that estrogens act directly on the nervous system to affect the severity of pain. Estrogen receptors (ERs) are expressed by sensory neurons, and in trigeminal ganglia, 17β-estradiol can indirectly enhance nociception by stimulating expression and release of prolactin, which increases phosphorylation of the nociceptor transducer transient receptor potential vanilloid receptor 1 (TRPV1). Here, we show that 17β-estradiol acts directly on dorsal root ganglion (DRG) sensory neurons to reduce TRPV1 activation by capsaicin. Capsaicin-induced cobalt uptake and the maximum TRPV1 current induced by capsaicin were inhibited when isolated cultured DRGs neurons from adult female rats were exposed to 17β-estradiol (10–100 nm) overnight. There was no effect of 17β-estradiol on capsaicin potency, TRPV1 activation by protons (pH 6–4), and P2X currents induced by α,β-methylene-ATP. Diarylpropionitrile (ERβ agonist) also inhibited capsaicin-induced TRPV1 currents, whereas propylpyrazole triol (ERα agonist) and 17α-estradiol (inactive analog) were inactive, and 17β-estradiol conjugated to BSA (membrane-impermeable agonist) caused a small increase. TRPV1 inhibition was antagonized by tamoxifen (1 μm), but ICI182870 (10 μm) was a potent agonist and mimicked 17β-estradiol. We conclude that TRPV1 in DRG sensory neurons can be inhibited by a nonclassical estrogen-signalling pathway that is downstream of intracellular ERβ. This affects the vanilloid binding site targeted by capsaicin but not the TRPV1 activation site targeted by protons. These actions could curtail the nociceptive transducer functions of TRPV1 and limit chemically induced nociceptor sensitization during inflammation. They are consistent with clinical reports that female pelvic pain can increase after reductions in circulating estrogens.
Publisher: Springer Science and Business Media LLC
Date: 11-04-2019
Publisher: Springer Science and Business Media LLC
Date: 11-2017
Publisher: Elsevier BV
Date: 02-2004
Publisher: Wiley
Date: 03-06-1996
DOI: 10.1002/(SICI)1096-9861(19960603)369:3<372::AID-CNE4>3.0.CO;2-2
Abstract: Outpatient or ambulatory treatment for prolonged air leak (PAL) has been reported previously in various studies. Evidence regarding efficiency and safety is nevertheless poor. This report describes the experience of 10 years ambulatory care with a digital chest drain system monitored by specialized nurses in our centre. The aim of the study is to give further insights in the effectiveness and safety of this treatment. Retrospective data of 10 years ambulatory care for PAL were examined. One hundred and forty patients with PAL after pneumothorax or pulmonary surgery were included. A total of 140 patients with PAL were included. Treatment was successful in 112 patients (80.0%). Hospital readmission was necessary in 33 patients (23.6%) and 28 (20.0%) of them received additional treatment. Additional treatment consisted of video-assisted thoracoscopic surgery (VATS) in 19 patients (13.6%), new chest tube placement in 8 patients (5.7%) and pleurodesis (with talc slurry) in 1 patient (0.7%). Minor complications occurred in 10 patients (7.1%), major complications requiring readmission occurred in 14 patients (10.0%). Ambulatory treatment of PAL with a digital monitoring device resulted in a high success rate with a limited complication rate.
Publisher: Elsevier BV
Date: 08-2007
Publisher: Springer Science and Business Media LLC
Date: 23-05-2006
Publisher: Elsevier BV
Date: 02-2018
Publisher: The Company of Biologists
Date: 07-2021
DOI: 10.1242/DMM.049068
Abstract: Benign prostatic hyperplasia/lower urinary tract dysfunction (LUTD) affects nearly all men. Symptoms typically present in the fifth or sixth decade and progressively worsen over the remainder of life. Here, we identify a surprising origin of this disease that traces back to the intrauterine environment of the developing male, challenging paradigms about when this disease process begins. We delivered a single dose of a widespread environmental contaminant present in the serum of most Americans [2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD), 1 µg/kg], and representative of a broader class of environmental contaminants, to pregnant mice and observed an increase in the abundance of a neurotrophic factor, artemin, in the developing mouse prostate. Artemin is required for noradrenergic axon recruitment across multiple tissues, and TCDD rapidly increases prostatic noradrenergic axon density in the male fetus. The hyperinnervation persists into adulthood, when it is coupled to autonomic hyperactivity of prostatic smooth muscle and abnormal urinary function, including increased urinary frequency. We offer new evidence that prostate neuroanatomical development is malleable and that intrauterine chemical exposures can permanently reprogram prostate neuromuscular function to cause male LUTD in adulthood.
Publisher: ZappyLab, Inc.
Date: 17-12-2019
DOI: 10.17504/PROTOCOLS.IO.BAKJICUN
Abstract: This protocol is used for bladder cannulation and cystometry in an experimental adult male or female rat. The surgery is performed under anesthesia and should incorporate all local requirements for standards of animal experimentation, including methods of anesthesia, surgical environment, and post-operative monitoring and care.
Publisher: American Physiological Society
Date: 2003
Abstract: Androgens have potent effects on the maturation and maintenance of a number of neural pathways involved in reproductive behaviors in males. Most studies in this area have focused on central pathways, but androgen receptors are expressed by many peripheral neurons innervating reproductive organs, and previous studies have demonstrated structural and chemical changes in these neurons at puberty and after castration. We have performed the first electrophysiological comparison of pelvic autonomic ganglion neurons in male rats before and after puberty and following pre- or postpubertal castration. Studies were performed in vitro on intact ganglia with hypogastric and pelvic nerves attached to allow synaptic activation of sympathetic or parasympathetic neurons, respectively. Pelvic ganglion neurons underwent many changes in their passive and active membrane properties over the pubertal period, and some of these changes were dependent on exposure to circulating androgens. The most pronounced steroid-dependent effects were on membrane capacitance (soma size) in sympathetic neurons and duration of the action potential afterhyperpolarization in tonic neurons. Our study also showed that rat pelvic ganglion cells and their synaptic inputs were more erse than previously reported. In conclusion, this study demonstrated that rat pelvic ganglion neurons undergo considerable postnatal changes in their electrophysiological properties. The steroid dependence of some of these changes indicates that circulating androgens may influence reproductive behaviors at many locations within the nervous system not just in the brain and spinal cord.
Publisher: Oxford University Press (OUP)
Date: 03-11-2020
Abstract: Do menstrual cycle-dependent changes occur in the histological appearance of superficial peritoneal endometriotic lesions, and are they equivalent to those observed in the eutopic endometrium? Only a small subset of superficial peritoneal endometriotic lesions exhibits some histological features in phase with menstrual cycle-related changes observed in eutopic endometrium. Endometriotic lesions are frequently described as implants that follow menstrual cycle-related changes in morphology, as per the eutopic endometrium. This concept has been widely accepted despite the lack of conclusive published evidence. This was a retrospective cohort study of 42 patients, from across the menstrual cycle, with surgically and histologically confirmed endometriosis. Patients were a subset selected from a larger endometriosis study being conducted at the Royal Women’s Hospital, Melbourne since 2012. Histological features of epithelium, stroma and gland morphology were examined in haematoxylin and eosin stained sections of superficial peritoneal endometriotic lesions and matched eutopic endometrium (menstrual: n = 4, proliferative: n = 11, secretory: n = 17, hormone-treated: n = 10). At least two biopsies (average = 4, range = 2–8 biopsies) and a matched endometrial s le were analysed for each patient and results were presented per endometriotic gland profile (n = 1051). Data were analysed using mixed effects logistic regression to account for multiple patients and multiple endometriotic biopsies, each with multiple endometriotic gland profiles. This model also enabled analysis of endometriotic lesions versus eutopic endometrium. There was considerable inter- and intra-patient variability in the morphology of superficial peritoneal endometriotic lesions. Menstrual cycle-associated changes were only observed for some features in a subset of endometriotic gland profiles. The proportion of endometriotic gland profiles with epithelial mitoses significantly increased in the proliferative phase (18% of gland profiles) relative to the menstrual phase (0% of endometriotic gland profiles) (odds ratios (OR) 9.30 95% confidence intervals (CI) = 3.71–23.32 P & 0.001). Fewer blood-filled gland lumens were observed in the secretory phase (45% of endometriotic gland profiles) compared to the menstrual phase (67% of endometriotic gland profiles) (OR, 0.30 95% CI = 0.11–0.79 P = 0.015). The features of the eutopic endometrium analysed in this study did not reflect the results in matched endometriotic lesions (P & 0.05). Not applicable. This study focused on features observed in sections of superficial peritoneal lesions and these may differ from features of deep infiltrating endometriosis or ovarian endometriomas. Cycle phases were limited to menstrual, proliferative and secretory phases to allow appropriate statistical modelling. This study highlights heterogeneity in the histological characteristics of superficial peritoneal lesions. It challenges the assumption that lesion morphology consistently reflects menstrual cycle-associated changes. Research reported in this publication was supported in part by National Health and Medical Research Council (NHMRC) project grants GNT1012245, GNT1105321 and GNT1026033 (P.A.W.R., J.E.G. and S.J.H.-C.). There are no competing interests.
Publisher: ZappyLab, Inc.
Date: 12-12-2021
DOI: 10.17504/PROTOCOLS.IO.B2SSQEEE
Abstract: This protocol is suitable for preserving tissues for ultrastructural neuroanatomical studies of peripheral nerves, ganglia, spinal cord or brain in adult rats. The protocol is performed under anesthesia and should incorporate all local requirements for standards of animal experimentation.
Publisher: Elsevier BV
Date: 03-2015
DOI: 10.1016/J.MCN.2015.03.004
Abstract: Neurotrophic factors have been intensively studied as potential therapeutic agents for promoting neural regeneration and functional recovery after nerve injury. Artemin is a member of the glial cell line-derived neurotrophic factor (GDNF) family of ligands (GFLs) that forms a signalling complex with GFRα3 and the tyrosine kinase Ret. Systemic administration of artemin in rodents is reported to facilitate regeneration of primary sensory neurons following axotomy, improve recovery of sensory function, and reduce sensory hypersensitivity that is a cause of pain. However, the biological mechanisms that underlie these effects are mostly unknown. This study has investigated the biological significance of the colocalisation of GFRα3 with TrkA (neurotrophin receptor for nerve growth factor [NGF]) in the peptidergic type of unmyelinated (C-fibre) sensory neurons in rat dorsal root ganglia (DRG). In vitro neurite outgrowth assays were used to study the effects of artemin and NGF by comparing DRG neurons that were previously uninjured, or were axotomised in vivo by transecting a visceral or somatic peripheral nerve. We found that artemin could facilitate neurite initiation but in comparison to NGF had low efficacy for facilitating neurite elongation and branching. This low efficacy was not increased when a preconditioning in vivo nerve injury was used to induce a pro-regenerative state. Neurite initiation was unaffected by artemin when PI3 kinase and Src family kinase signalling were blocked, but NGF had a reduced effect.
Publisher: ZappyLab, Inc.
Date: 13-01-2019
DOI: 10.17504/PROTOCOLS.IO.W2XFGFN
Abstract: This protocol is used to visualise sensory and autonomic neurons innervating the bladder body (dome), bladder trigone or proximal urethra in an experimental adult male or female rat. The protocol is performed under anesthesia and should incorporate all local requirements for standards of animal experimentation, including methods of anesthesia, surgical environment, and post-operative monitoring and care.
Publisher: MDPI AG
Date: 13-04-2023
Abstract: Environmental pollution has increased over the past few decades, posing serious risks to all biological systems, including the poultry sector. Oxidative stress in chickens caused by dietary, environmental, and pathological variables influences how well chickens perform as well as the quality of meat and eggs. Lead (Pb) and cadmium (Cd) are two ex les of heavy metals that are harmful for chicken health. They can cause oxidative stress by increasing the production of reactive oxygen species (ROS) and reactive nitrogen species (RNS) and blocking antioxidants from protecting cells from increased amounts of free radicals. The oxidative state of heavy metals, their interactions with endogenous antioxidants, and chemical processes all affect how hazardous they are to the body. Today, scientists have investigated and applied a variety of nutritional tactics to lessen the harmful effects of oxidative stress on animal health brought on by heavy metals. Researchers have recently become interested in the chemicals because of their chelating and growth-stimulating functions, as well as the antioxidant qualities of useful plant components. The deleterious consequences of oxidative stress induced by two heavy metals on chickens is discussed in this review, along with phytogenic use as a potential intervention strategy to lessen these effects and maintain the redox equilibrium in poultry.
Publisher: Wiley
Date: 06-2006
Location: Iran (Islamic Republic of)
No related grants have been discovered for Janet Keast.