ORCID Profile
0000-0001-9189-4430
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Publisher: Elsevier BV
Date: 12-1988
DOI: 10.1016/0165-0270(88)90166-5
Abstract: The aim of this study, performed on anaesthetized cats and rabbits, was to test the assumption that pressure microinjections of excitatory amino acids cause long-lasting excitation of neurones located close to the injection site. Unitary action potentials or antidromic field potentials were recorded from respiratory or 'reticular' neurones in the medulla oblongata and from phrenic motoneurones at different distances from the injection site. Injection of 10-150 nl (5-150 nmol) of L-glutamate or DL-homocysteic acid into these areas resulted in complex and widespread neuronal events. Generally, more distant neurones (500-1300 microns) were excited for variable periods of time (3-15 min), while neurones in the vicinity of the injection site (0-500 microns) showed, after a brief period of excitation time, a long-lasting (up to 30 min) decrease in excitability or silencing of discharge, probably due to a depolarizing block and disturbances in the ionic composition of the extracellular space. These findings show that interpretation of physiological responses following such injections should not be based on an assumption of local neuronal excitation. Some recommendations regarding the use of this technique are made.
Publisher: Elsevier BV
Date: 05-1999
DOI: 10.1016/S0896-6273(00)80762-X
Abstract: Several distinct mechanisms may cause synaptic depression, a common form of short-term synaptic plasticity. These include postsynaptic receptor desensitization, presynaptic depletion of releasable vesicles, or other presynaptic mechanisms depressing vesicle release. At the endbulb of Held, a fast central calyceal synapse in the auditory pathway, cyclothiazide (CTZ) abolished marked paired pulse depression (PPD) by acting presynaptically to enhance transmitter release, rather than by blocking postsynaptic receptor desensitization. PPD and its response to CTZ were not altered by prior depletion of the releasable vesicle pool but were blocked by lowering external calcium concentration, while raising external calcium enhanced PPD. We conclude that a major component of PPD at the endbulb is due to a novel, transient depression of release, which is dependent on the level of presynaptic calcium entry and is CTZ sensitive.
Publisher: Springer Science and Business Media LLC
Date: 11-09-2010
DOI: 10.1007/S10735-010-9293-3
Abstract: Impairment of the blood-brain barrier (BBB), the blood-cerebrospinal fluid (CSF) barrier and brain-CSF barrier has been implicated in neuropathology of several brain disorders, such as amyotrophic lateral sclerosis, cerebral edema, multiple sclerosis, neural inflammation, ischemia and stroke. Two-pore domain weakly inward rectifying K+ channel (TWIK)-related acid-sensitive potassium (TASK)-1 channels (K2p3.1 KCNK3) are among the targets that contribute to the development of these pathologies. For ex le TASK-1 activity is inhibited by acidification, ischemia, hypoxia and several signaling molecules released under pathologic conditions. We have used immuno-histochemistry to examine the distribution of the TASK-1 protein in structures associated with the BBB, blood-CSF barrier, brain-CSF barrier, and in the meninges of adult rat. Dense TASK-1 immuno-reactivity (TASK-1-IR) was observed in ependymal cells lining the fourth ventricle at the brain-CSF interface, in glial cells that ensheath the walls of blood vessels at the glio-vascular interface, and in the meninges. In these structures, TASK-1-IR often co-localized with glial fibrillary associated protein (GFAP) or vimentin. This study provides anatomical evidence for localization of TASK-1 K+ channels in cells that segregate distinct fluid compartments within and surrounding the brain. We suggest that TASK-1 channels, in coordination with other ion channels (e.g., aquaporins and chloride channels) and transporters (e.g., Na+-K+-ATPase and Na+-K+-2Cl⁻ and by virtue of its heterogeneous distribution, may differentially contribute to the varying levels of K+ vital for cellular function in these compartments. Our findings are likely to be relevant to recently reported roles of TASK-1 in cerebral ischemia, stroke and inflammatory brain disorders.
Publisher: Wiley
Date: 02-11-2020
DOI: 10.1002/AR.24542
Abstract: The total motor neuron (MN) somato‐dendritic surface area is correlated with motor unit type. MNs with smaller surface areas innervate slow (S) and fast fatigue‐resistant (FR) motor units, while MNs with larger surface areas innervate fast fatigue‐intermediate (FInt) and fast fatigable (FF) motor units. Differences in MN surface area (equivalent to membrane capacitance) underpin the intrinsic excitability of MNs and are consistent with the orderly recruitment of motor units (S FR FInt FF) via the Size Principle. In amyotrophic lateral sclerosis (ALS), large MNs controlling FInt and FF motor units exhibit earlier denervation and death, compared to smaller and more resilient MNs of type S and FR motor units that are spared until late in ALS. Abnormal dendritic morphologies in MNs precede neuronal death in human ALS and in rodent models. We employed Golgi‐Cox methods to investigate somal size‐dependent changes in the dendritic morphology of hypoglossal MNs in wildtype and SOD1 G93A mice (a model of ALS), at postnatal (P) day ~30 (pre‐symptomatic), ~P60 (onset), and ~P120 (mid‐disease) stages. In wildtype hypoglossal MNs, increased MN somal size correlated with increased dendritic length and spines in a linear fashion. By contrast, in SOD1 G93A mice, significant deviations from this linear correlation were restricted to the larger vulnerable MNs at pre‐symptomatic (maladaptive) and mid‐disease (degenerative) stages. These findings are consistent with excitability changes observed in ALS patients and in rodent models. Our results suggest that intrinsic or synaptic increases in MN excitability are likely to contribute to ALS pathogenesis, not compensate for it.
Publisher: Springer Science and Business Media LLC
Date: 26-11-2015
DOI: 10.1007/S00429-013-0673-9
Abstract: During mid to late embryonic development (E13 to birth in mice), the neuromotor system is refined by reducing motor neuron (MN) numbers and establishing nascent synaptic connections onto and by MNs. Concurrently, the response to GABAergic and glycinergic synaptic activity switches from postsynaptic excitation to inhibition. Our previous studies on mutant mice lacking glycinergic transmission or deficient in GABA suggests that altered MN activity levels during this developmental period differentially regulates MN survival and muscle innervation for respiratory and non-respiratory motor pools. To determine if combined loss of GABAergic and glycinergic transmission plays a similar or exaggerated role, we quantified MN number and muscle innervation in two respiratory (hypoglossal and phrenic) and two locomotor (brachial and lumbar) motor pools, in mice lacking vesicular inhibitory amino acid transporter, which display absent or severely impaired GABAergic and glycinergic neurotransmission. For respiratory MNs, we observed significant decreases in MN number (-20 % hypoglossal and -36 % phrenic) and diaphragm axonal branching (-60 %). By contrast, for non-respiratory brachial and lumbar MNs, we observed increases in MN number (+62 % brachial and +84 % lumbar) and axonal branching for innervated muscles (+123 % latissimus dorsi for brachial and +61 % gluteal for lumbar). These results show that combined absence of GABAergic and glycinergic neurotransmission causes distinct regional changes in MN number and muscle innervation, which are dependent upon the motor function of the specific motor pool.
Publisher: Springer Science and Business Media LLC
Date: 14-09-2006
Publisher: Society for Neuroscience
Date: 06-01-2016
DOI: 10.1523/JNEUROSCI.1576-15.2016
Abstract: Emerging evidence suggests that central synaptic inputs onto motor neurons (MNs) play an important role in developmental regulation of the final number of MNs and their muscle innervation for a particular motor pool. Here, we describe the effect of genetic deletion of glycinergic neurotransmission on single MN structure and on functional excitatory and inhibitory inputs to MNs. We measured synaptic currents in E18.5 hypoglossal MNs from brain slices using whole-cell patch-cl recording, followed by dye-filling these same cells with Neurobiotin, to define their morphology by high-resolution confocal imaging and 3D reconstruction. We show that hypoglossal MNs of mice lacking gephyrin display increased dendritic arbor length and branching, increased spiny processes, decreased inhibitory neurotransmission, and increased excitatory neurotransmission. These findings suggest that central glycinergic synaptic activity plays a vital role in regulating MN morphology and glutamatergic central synaptic inputs during late embryonic development. SIGNIFICANCE STATEMENT MNs within the brainstem and spinal cord are responsible for integrating a erse array of synaptic inputs into discrete contractions of skeletal muscle to achieve coordinated behaviors, such as breathing, vocalization, and locomotion. The last trimester in utero is critical in neuromotor development, as this is when central and peripheral synaptic connections are made onto and from MNs. At this time-point, using transgenic mice with negligible glycinergic postsynaptic responses, we show that this deficiency leads to abnormally high excitatory neurotransmission and alters the dendritic architecture responsible for coherently integrating these inputs. This study compliments the emerging concept that neurodevelopmental disorders (including autism, epilepsy, and amyotrophic lateral sclerosis) are underpinned by synaptic dysfunction and therefore will be useful to neuroscientists and neurologists alike.
Publisher: Society for Neuroscience
Date: 22-10-2008
DOI: 10.1523/JNEUROSCI.1340-08.2008
Abstract: Distinguishing the primary from secondary effects and compensatory mechanisms is of crucial importance in understanding adult-onset neurodegenerative diseases such as amyotrophic lateral sclerosis (ALS). Transgenic mice that overexpress the G93A mutation of the human Cu-Zn superoxide dismutase 1 gene (hSOD1 G93A mice) are a commonly used animal model of ALS. Whole-cell patch-cl recordings from neurons in acute slice preparations from neonatal wild-type and hSOD1 G93A mice were made to characterize functional changes in neuronal activity. Hypoglossal motoneurons (HMs) in postnatal day 4 (P4)–P10 hSOD1 G93A mice displayed hyperexcitability, increased persistent Na + current (PC Na ), and enhanced frequency of spontaneous excitatory and inhibitory transmission, compared with wild-type mice. These functional changes in neuronal activity are the earliest yet reported for the hSOD1 G93A mouse, and are present 2–3 months before motoneuron degeneration and clinical symptoms appear in these mice. Changes in neuronal activity were not restricted to motoneurons: superior colliculus interneurons also displayed hyperexcitability and synaptic changes (P10–P12). Furthermore, in vivo viral-mediated GFP (green fluorescent protein) overexpression in hSOD1 G93A HMs revealed precocious dendritic remodeling, and behavioral assays revealed transient neonatal neuromotor deficits compared with controls. These findings underscore the widespread and early onset of abnormal neural activity in this mouse model of the adult neurodegenerative disease ALS, and suggest that suppression of PC Na and hyperexcitability early in life might be one way to mitigate or prevent cell death in the adult CNS.
Publisher: American Society for Pharmacology & Experimental Therapeutics (ASPET)
Date: 09-06-2005
Abstract: Tertiapin, a short peptide from honey bee venom, has been reported to specifically block the inwardly rectifying K(+) (Kir) channels, including G protein-coupled inwardly rectifying potassium channel (GIRK) 1+GIRK4 heteromultimers and ROMK1 homomultimers. In the present study, the effects of a stable and functionally similar derivative of tertiapin, tertiapin-Q, were examined on recombinant human voltage-dependent Ca(2+)-activated large conductance K(+) channel (BK or MaxiK alpha-subunit or hSlo1 homomultimers) and mouse inwardly rectifying GIRK1+GIRK2 (i.e., Kir3.1 and Kir3.2) heteromultimeric K(+) channels expressed in Xenopus oocytes and in cultured newborn mouse dorsal root ganglion (DRG) neurons. In two-electrode voltage-cl ed oocytes, tertiapin-Q (1-100 nM) inhibited BK-type K(+) channels in a use- and concentration-dependent manner. We also confirmed the inhibition of recombinant GIRK1+GIRK2 heteromultimers by tertiapin-Q, which had no effect on endogenous depolarization- and hyperpolarization-activated currents sensitive to extracellular alent cations (Ca(2+), Mg(2+), Zn(2+), and Ba(2+)) in defolliculated oocytes. In voltage-cl ed DRG neurons, tertiapin-Q voltage- and use-dependently inhibited outwardly rectifying K(+) currents, but Cs(+)-blocked hyperpolarization-activated inward currents including I(H) were insensitive to tertiapin-Q, baclofen, barium, and zinc, suggesting absence of functional GIRK channels in the newborn. Under current-cl conditions, tertiapin-Q blocked the action potential after hyperpolarization (AHP) and increased action potential duration in DRG neurons. Taken together, these results demonstrate that the blocking actions of tertiapin-Q are not specific to Kir channels and that the blockade of recombinant BK channels and native neuronal AHP currents is use-dependent. Inhibition of specific types of Kir and voltage-dependent Ca(2+)-activated K(+) channels by tertiapin-Q at nanomolar range via different mechanisms may have implications in pain physiology and therapy.
Publisher: Elsevier BV
Date: 05-2006
DOI: 10.1016/J.NEULET.2006.01.051
Abstract: We have compared the expression pattern of NMDA receptor subunits (NR1 and NR2A-D) and NR1 splice variants (NR1-1a/1b,-2a/2b,-3a/3b,-4a/4b) in motor neuron populations from adult Wistar rats that are vulnerable (hypoglossal, XII) or resistant (oculomotor, III) to death in amyotrophic lateral sclerosis (ALS). The major finding was higher levels of expression of the NR2B subunit in the hypoglossal nucleus. Quantitative real-time PCR showed that NR1 was expressed at a greater level than any of the NR2 subunits (>15 fold greater, P<or=0.05, n = 11 animals), while conventional RT-PCR showed no difference in NR1 splice variant expression (with all variants except NR1-3 detected in both nuclei n = 6 animals). Within III, the NR2B subunit was expressed 1.7 to 2.6-fold lower than the other NR2 subunits (P<or=0.05), while in XII all NR2 subunits were expressed at equal levels. When comparing levels between the 2 nuclei, mRNA for the NR2B subunit was expressed 2.1-fold higher in XII compared to III (P<or=0.05), while their was no difference in mRNA expression for the other subunits. Immunohistochemical analysis confirmed greater NR2B protein levels within in idual hypoglossal neurons compared to oculomotor neurons (1.8-fold greater, P<or=0.05, n = 5 animals). Lower expression of the NMDA NR2B subunit may constitute one factor conferring protection to oculomotor neurons in ALS.
Publisher: Elsevier BV
Date: 11-1990
DOI: 10.1016/0006-8993(90)91807-S
Abstract: Extracellular recordings were made in the C5 segment of the spinal cord of anaesthetised cats from 129 units which showed respiratory phased discharge. The majority of recordings (88%) were thought to arise from the somata of respiratory spinal interneurons. Inspiratory units and expiratory units comprised 42% and 52% of all recorded units. A small number of postinspiratory units were also found (n = 5). Most units did not respond to electrical stimulation of the ipsilateral superior laryngeal (SLN) and phrenic nerves (PN), but a few expiratory (n = 2) and postinspiratory units (n = 1) were excited by SLN stimulation, while 6 inspiratory units had their discharge suppressed by the same stimulus. PN stimulation evoked a long latency (2-7 ms) burst of firing in 2 inspiratory and 1 expiratory interneurons. It is concluded that these respiratory interneurons may provide a segmental input to phrenic motoneurons, in addition to synaptic drives mediated by bulbospinal pathways.
Publisher: Wiley
Date: 10-1998
DOI: 10.1111/J.1440-1681.1998.TB02166.X
Abstract: 1. The central pattern generator (CPG) for respiration is located in the brainstem and produces rhythmic synaptic drive for motoneurons controlling respiratory muscles. Based on respiratory nerve discharge, the respiratory cycle can be ided into three phases: inspiration, postinspiration and stage 2 expiration. 2. Six basic types of respiratory neuron participate in respiratory rhythmogenesis. Their firing and membrane potential patterns are locked to different phases of the respiratory cycle. 3. In adult mammals, respiratory neurons are subject to excitatory and inhibitory synaptic inputs and show extensive synaptic interconnections that are mainly inhibitory. There are differences in the relative importance of excitatory and inhibitory synaptic drives and the neurotransmitters involved in respiratory rhythmogenesis in neonates compared with adults. 4. Respiratory neurons possess a number of intrinsic membrane currents that may be involved in central pattern generation, including low- and high-voltage-activated calcium, potassium, calcium-dependent potassium, sodium and mixed cationic currents. More quantitative information is needed about the distribution and characteristics of these ionic currents if we are to understand rhythmogenesis. 5. The two main theories for the origin of respiratory rhythm are those of pacemaker neuron-driven and synaptic network-driven CPG. Evidence derived from in vivo and in vitro experiments exists to support both of these theories. There may be a significant switch in the underlying mechanism driving the respiratory CPG during postnatal development.
Publisher: Elsevier BV
Date: 02-1994
DOI: 10.1016/0006-8993(94)91025-1
Abstract: The pontine respiratory neurons (PRG) in the 'pneumotaxic centre' have been hypothesized to contribute to phase-switching of neural respiratory activity, especially in terminating inspiration. To define the neural elements involved in phase-switching, we recorded respiratory neurons extra- and intracellularly in anesthetized cats with an intact central nervous system. In total, 54 neurons were recorded: 49 neurons with activity modulated by central respiratory rhythm (20 inspiratory, 17 postinspiratory and 12 expiratory) and 5 neurons with activity correlated to tracheal pressure. The recorded neurons were clustered in dorsolateral pontine tegmentum within the Kölliker-Fuse (KF) subnucleus of the parabrachial nuclei. Stable intracellular membrane potential was recorded in 11 of the 49 respiratory neurons (8 postinspiratory, 1 early inspiratory and 2 inspiratory). During continuous injection of chloride ions (n = 6), synaptic noise increased and IPSPs reversed, including a wave of IPSPs during stage-2 expiration in postinspiratory neurons. Further, relative input resistance varied through the respiratory cycle such that the least input resistance occurred during the neuron's (n = 5) quiescent period. No IPSPs nor EPSPs were evoked in pontine respiratory neurons by vagal stimulation. In conclusion, various types of respiratory neurons were recorded in the KF nucleus. Prominent excitatory and inhibitory postsynaptic activities were similar to those described for medullary neurons. These pontine respiratory neurons do not appear to receive a strong afferent input from the vagus. Rather, vagal afferent inputs seem to be directed towards non-respiratory neurons that are located more medially in the dorsal pons.
Publisher: Elsevier BV
Date: 10-2012
DOI: 10.1016/J.CLINPH.2012.01.028
Abstract: To assess the relationship between Bayesian MUNE and histological motor neuron counts in wild-type mice and in an animal model of ALS. We performed Bayesian MUNE paired with histological counts of motor neurons in the lumbar spinal cord of wild-type mice and transgenic SOD1(G93A) mice that show progressive weakness over time. We evaluated the number of acetylcholine endplates that were innervated by a presynaptic nerve. In wild-type mice, the motor unit number in the gastrocnemius muscle estimated by Bayesian MUNE was approximately half the number of motor neurons in the region of the spinal cord that contains the cell bodies of the motor neurons supplying the hindlimb crural flexor muscles. In SOD1(G93A) mice, motor neuron numbers declined over time. This was associated with motor endplate denervation at the end-stage of disease. The number of motor neurons in the spinal cord of wild-type mice is proportional to the number of motor units estimated by Bayesian MUNE. In SOD1(G93A) mice, there is a lower number of estimated motor units compared to the number of spinal cord motor neurons at the end-stage of disease, and this is associated with disruption of the neuromuscular junction. Our finding that the Bayesian MUNE method gives estimates of motor unit numbers that are proportional to the numbers of motor neurons in the spinal cord supports the clinical use of Bayesian MUNE in monitoring motor unit loss in ALS patients.
Publisher: MDPI AG
Date: 27-04-2021
Abstract: Amyotrophic lateral sclerosis (ALS) is a progressive neuromotor disease characterized by the loss of upper and lower motor neurons (MNs), resulting in muscle paralysis and death. Early cortical hyper-excitability is a common pathological process observed clinically and in animal disease models. Although the mechanisms that underlie cortical hyper-excitability are not completely understood, the molecular and cellular mechanisms that cause enhanced neuronal intrinsic excitability and changes in excitatory and inhibitory synaptic activity are starting to emerge. Here, we review the evidence for an anterograde glutamatergic excitotoxic process, leading to cortical hyper-excitability via intrinsic cellular and synaptic mechanisms and for the role of interneurons in establishing disinhibition in clinical and experimental settings. Understanding the mechanisms that lead to these complex pathological processes will likely produce key insights towards developing novel therapeutic strategies to rescue upper MNs, thus alleviating the impact of this fatal disease.
Publisher: American Physiological Society
Date: 08-2015
Abstract: The basolateral amygdala (BLA) is a complex brain region associated with processing emotional states, such as fear, anxiety, and stress. Some aspects of these emotional states are driven by the network activity of synaptic connections, derived from both local circuitry and projections to the BLA from other regions. Although the synaptic physiology and general morphological characteristics are known for many in idual cell types within the BLA, the combination of morphological, electrophysiological, and distribution of neurochemical GABAergic synapses in a three-dimensional neuronal arbor has not been reported for single neurons from this region. The aim of this study was to assess differences in morphological characteristics of BLA principal cells and interneurons, quantify the distribution of GABAergic neurochemical synapses within the entire neuronal arbor of each cell type, and determine whether GABAergic synaptic density correlates with electrophysiological recordings of inhibitory postsynaptic currents. We show that BLA principal neurons form complex dendritic arborizations, with proximal dendrites having fewer spines but higher densities of neurochemical GABAergic synapses compared with distal dendrites. Furthermore, we found that BLA interneurons exhibited reduced dendritic arbor lengths and spine densities but had significantly higher densities of putative GABAergic synapses compared with principal cells, which was correlated with an increased frequency of spontaneous inhibitory postsynaptic currents. The quantification of GABAergic connectivity, in combination with morphological and electrophysiological measurements of the BLA cell types, is the first step toward a greater understanding of how fear and stress lead to changes in morphology, local connectivity, and/or synaptic reorganization of the BLA.
Publisher: Society for Neuroscience
Date: 14-01-2015
DOI: 10.1523/JNEUROSCI.3483-14.2015
Abstract: Motor cortex layer V pyramidal neurons (LVPNs) regulate voluntary control of motor output and selectively degenerate (along with lower motor neurons) in amyotrophic lateral sclerosis. Using dye-filling and whole-cell patch cl ing in brain slices, together with high-resolution spinning disk confocal z -stack mosaics, we characterized the earliest presymptomatic cortical LVPN morphologic and electrophysiological perturbations in hSOD1 G93A (SOD1) mice to date. Apical dendritic regression occurred from postnatal day (P) 28, dendritic spine loss from P21, and increased EPSC frequency from P21 in SOD1 LVPNs. These findings demonstrate extensive early changes in motor cortex of the SOD1 mouse model, which thus recapitulates clinically relevant cortical pathophysiology more faithfully than previously thought.
Publisher: Society for Neuroscience
Date: 02-02-2005
DOI: 10.1523/JNEUROSCI.1786-04.2005
Abstract: GABAergic and glycinergic synaptic transmission is proposed to promote the maturation and refinement of the developing CNS. Here we provide morphological and functional evidence that glycinergic and GABAergic synapses control motoneuron development in a region-specific manner during programmed cell death. In gephyrin-deficient mice that lack all postsynaptic glycine receptor and some GABA A receptor clusters, there was increased spontaneous respiratory motor activity, reduced respiratory motoneuron survival, and decreased innervation of the diaphragm. In contrast, limb-innervating motoneurons showed decreased spontaneous activity, increased survival, and increased innervation of their target muscles. Both GABA and glycine increased limb-innervating motoneuron activity and decreased respiratory motoneuron activity in wild-type mice, but only glycine responses were abolished in gephyrin-deficient mice. Our results provide genetic evidence that the development of glycinergic and GABAergic synaptic inputs onto motoneurons plays an important role in the survival, axonal branching, and spontaneous activity of motoneurons in developing mammalian embryos.
Publisher: MDPI AG
Date: 19-07-2022
DOI: 10.3390/IJMS23147965
Abstract: Normal development and function of the central nervous system involves a balance between excitatory and inhibitory neurotransmission. Activity of both excitatory and inhibitory neurons is modulated by inhibitory signalling of the GABAergic and glycinergic systems. Mechanisms that regulate formation, maturation, refinement, and maintenance of inhibitory synapses are established in early life. Deviations from ideal excitatory and inhibitory balance, such as down-regulated inhibition, are linked with many neurological diseases, including epilepsy, schizophrenia, anxiety, and autism spectrum disorders. In the mammalian forebrain, GABA is the primary inhibitory neurotransmitter, binding to GABA receptors, opening chloride channels and hyperpolarizing the cell. We review the involvement of down-regulated inhibitory signalling in neurological disorders, possible mechanisms for disease progression, and targets for therapeutic intervention. We conclude that transgenic models of disrupted inhibitory signalling—in GAD67+/− and VGAT−/− mice—are useful for investigating the effects of down-regulated inhibitory signalling in a range of neurological diseases.
Publisher: Frontiers Media SA
Date: 11-2017
Publisher: Springer Science and Business Media LLC
Date: 27-03-2011
DOI: 10.1007/S00232-011-9359-5
Abstract: Calcium-activated chloride currents (CaCCs) are required for epithelial electrolyte and fluid secretion, fertilization, sensory transduction and excitability of neurons and smooth muscle. Defolliculated Xenopus oocytes express a robust CaCC formed by a heterologous group of proteins including transmembrane protein 16A (TMEM16A) and bestrophins. Penetratin, a 17-amino acid peptide, potentiated endogenous oocyte CaCCs by ~50-fold at 10 μM, recorded using a two-electrode voltage cl . CaCC potentiation was rapid and dose-dependent (EC50=3.2 μM). Penetratin-potentiated currents reversed at -18 mV and were dependent on the extracellular alent cations present, showing positive regulation by Ca2+ and Mg2+ but effective block by Zn2+ (IC50=5.9 μM). Extracellular Cd2+, Cu2+ and Ba2+ resulted in bimodal responses: CaCC inhibition at low but potentiation at high concentrations. Intracellular BAPTA injection, which prevents activation of CaCCs, and the Cl- channel blockers niflumic acid and DIDS significantly reduced potentiation. In contrast, the K+ channel blockers Cs+, TEA, tertiapin-Q and halothane had no significant effect. This pharmacological profile is consistent with penetratin potentiation of zinc-sensitive CaCCs that are activated by influx of extracellular Ca2+. These findings may stimulate basic research on CaCCs in native cells and may lead to development of novel therapeutics targeting disorders caused by insufficient chloride secretion.
Publisher: The Endocrine Society
Date: 12-2013
DOI: 10.1210/EN.2013-1570
Abstract: GH deficiency is thought to be involved in the pathogenesis of amyotrophic lateral sclerosis (ALS). However, therapy with GH and/or IGF-I has not shown benefit. To gain a better understanding of the role of GH secretion in ALS pathogenesis, we assessed endogenous GH secretion in wild-type and hSOD1G93A mice throughout the course of ALS disease. Male wild-type and hSOD1G93A mice were studied at the presymptomatic, onset, and end stages of disease. To assess the pathological features of disease, we measured motor neuron number and neuromuscular innervation. We report that GH secretion profile varies at different stages of disease progression in hSOD1G93A mice compared with age-matched controls, GH secretion is unchanged prior to the onset of disease symptoms, elevated at the onset of disease symptoms, and reduced at the end stage of disease. In hSOD1G93A mice at the onset of disease, GH secretion is positively correlated with the percentage of neuromuscular innervation but not with motor neuron number. Moreover, this occurs in parallel with an elevation in the expression of muscle IGF-I relative to controls. Our data imply that increased GH secretion at symptom onset may be an endogenous endocrine response to increase the local production of muscle IGF-I to stimulate reinnervation of muscle, but that in the latter stages of disease this response no longer occurs.
Publisher: Frontiers Media SA
Date: 22-03-2019
Publisher: Elsevier BV
Date: 2004
Publisher: Wiley
Date: 09-1998
DOI: 10.1111/J.1469-7793.1998.861BG.X
Abstract: 1. Developmental changes in litude and time course of single-fibre-evoked and spontaneous EPSCs mediated by AMPA and NMDA receptors at the endbulb-bushy cell synapse of rats from 4 to 22 days of age were recorded using whole-cell patch-cl methods in in vitro slices of cochlear nucleus. 2. The mean conductance of the AMPA component of evoked EPSCs increased by 66 %, while that of the NMDA component decreased by 61 %, for 12- to 18-day-old rats cf. 4- to 11-day-old rats. 3. The mean AMPA spontaneous EPSC conductance increased by 54 %, while mean NMDA spontaneous EPSC conductance decreased by 83 %, for 12- to 22-day-old rats cf. 4- to 11-day-old rats. The mean number of quanta contributing to peak evoked AMPA conductance also increased by 78 % in the older age group, after correction for the asynchrony of evoked quantal release. 4. The decay time constant of spontaneous AMPA EPSCs showed a small decrease in older animals, while the decay time constant of spontaneous NMDA EPSCs was markedly decreased in older animals. The decay time constants of evoked NMDA EPSCs showed a quantitatively similar decrease to that of spontaneous NMDA EPSCs. This suggests that AMPA receptor subunit composition is unlikely to undergo developmental change, while NMDA receptor subunit composition may be substantially altered during synaptic maturation. 5. These data are consistent with a developmentally increased efficacy of AMPA receptor-mediated synaptic transmission at the endbulb-bushy cell synapse, due to an increase in underlying AMPA-mediated quantal size and content during the same period as a transient co-localization of NMDA receptors.
Publisher: Ovid Technologies (Wolters Kluwer Health)
Date: 03-2004
DOI: 10.1097/00001756-200403010-00011
Abstract: Developmental expression of two-pore domain potassium (2P K) channels, TASK-1 and TREK-1, was investigated in the rat cochlea at onset of hearing and after maturity using RT-PCR and immunocytochemistry. TASK-1 and TREK-1 mRNAs were detected by RT-PCR at postnatal day (P) 9-12. TASK-1 like immunoreactivity (LIR) in the P13 cochlea was observed in Deiters', pillar, Claudius' and outer sulcus cells, spiral limbus fibrocytes, and neuroglia. At P13, TREK-1-LIR was more wide-spread, and included sensory and supporting cells of the organ of Corti, spiral ganglion, stria vascularis, Reissner's membrane, inner and outer sulcus cells, connective and support tissues surrounding modiolus. By P105 the pattern of TASK-1- and TREK-1-LIR became limited to a subset of the above structures, suggesting developmental regulation. During postnatal development, TASK-1 may be important in the onset (around P11) and maturation (by P22) of endocochlear potential and hearing. The distribution of TASK-1 and TREK-1 suggest a role in K cycling and homeostasis. As TASK-1 and TREK-1 are inhibited by local anesthetics at doses used to treat tinnitus, 2P K channels may also be important in cochlear dysfunction.
Publisher: University of Toronto Press Inc. (UTPress)
Date: 07-2007
Abstract: There is a tendency for students from different nationalities to remain within groups of similar cultural backgrounds. The study reported here used group project work to encourage integration and cooperative learning between Australian students and Asian (Southeast Asian) international students in the second year of a veterinary science program. The group project involved an oral presentation during a second-year course (Structure and Function), with group formation engineered to include very high, high, moderate, and low achievers (based on previous grades). One Asian student and three Australian students were placed in each group. Student perceptions of group dynamics were analyzed through a self-report survey completed at the end of the presentations and through group student interviews. Results from the survey were analyzed by chi-square to compare the responses between Asian and Australian students, with statistical significance accepted at p 0.05. There were too few Asian students for statistical analysis from a single year therefore, the results from two successive years, 2004 (N = 104 26% Asian) and 2005 (N = 105 20% Asian), were analyzed. All participating students indicated in the interviews that the project was worthwhile and a good learning experience. Asian students expressed a greater preference for working in a group than for working alone (p = 0.001) and reported more frequently than Australian students that teamwork produces better results (p = 0.01). Australian students were more likely than Asian students to voice their opinion in a team setting (p = 0.001), while Asian students were more likely to depend on the lecturer for directions (p = 0.001). The results also showed that group project work appeared to create an environment that supported learning and was a successful strategy to achieve acceptance of cultural differences.
Publisher: Springer Science and Business Media LLC
Date: 04-08-2016
Publisher: Wiley
Date: 04-01-2000
DOI: 10.1046/J.1440-1681.2000.03192.X
Abstract: 1. Shifts in behavioural state are controlled by reciprocal changes in discharge of cholinergic and aminergic groups of brain‐stem ontine neurons. During rapid eye movement (REM) sleep, cholinergic neurons are most active and aminergic neurons are least active. 2. Significant changes occur in the central control of breathing during REM sleep respiration rate increases in frequency and variability, brain‐stem respiratory neuron discharge is generally enhanced and the outputs of some respiratory motor neuron pools are depressed. 3. Hypoglossal motor neurons (HM) control tongue movement and their depression during REM sleep has been implicated in obstructive sleep apnoea. The cellular basis of HM depression has been investigated in vitro and may be due to enhanced activation of cholinergic receptors or decreased activation of aminergic receptors. 4. In vitro preparations that show respiratory rhythmogenesis possess advantages for the investigation of the neurochemical basis of state‐dependent changes in respiration. Cholinergic changes in respiratory modulation of HM recorded in rhythmic brain‐stem slices from mice depend on the site of activation of cholinergic receptors.
Publisher: Walter de Gruyter GmbH
Date: 1993
Publisher: MDPI AG
Date: 19-12-2017
Publisher: Wiley
Date: 17-01-2011
Publisher: Elsevier BV
Date: 03-2016
DOI: 10.1016/J.EXPNEUROL.2016.12.004
Abstract: There is an emerging body of evidence that glycinergic and GABAergic synaptic inputs onto motor neurons (MNs) help regulate the final number of MNs and axonal muscle innervation patterns. Using mutant glutamate decarboxylase 67 (GAD67) and vesicular inhibitory amino acid transporter (VGAT) deficient mice, we describe the effect that deficiencies of presynaptic GABAergic and/or glycinergic release have on the post-synaptic somato-dendritic structure of motor neurons, and the development of excitatory and inhibitory synaptic inputs to MNs. We use whole-cell patch cl recording of synaptic currents in E18.5 hypoglossal MNs from brainstem slices, combined with dye-filling of these recorded cells with Neurobiotin™, high-resolution confocal imaging and 3-dimensional reconstructions. Hypoglossal MNs from GAD67- and VGAT-deficient mice display decreased inhibitory neurotransmission and increased excitatory synaptic inputs. These changes are associated with increased dendritic arbor length, increased complexity of dendritic branching, and increased density of spiny processes. Our results show that presynaptic release of inhibitory amino acid neurotransmitters are potent regulators of hypoglossal MN morphology and key regulators of synaptic inputs during this critical developmental time point.
Publisher: Elsevier BV
Date: 07-2002
DOI: 10.1016/S1569-9048(02)00043-5
Abstract: Respiration is altered during different stages of the sleep-wake cycle. We review the contribution of cholinergic systems to this alteration, with particular reference to the role of muscarinic acetylcholine receptors (MAchRs) during rapid eye movement (REM) sleep. Available evidence demonstrates that MAchRs have potent excitatory effects on medullary respiratory neurones and respiratory motoneurones, and are likely to contribute to changes in central chemosensitive drive to the respiratory control system. These effects are likely to be most prominent during REM sleep, when cholinergic brainstem neurones show peak activity levels. It is possible that MAchR dysfunction is involved in sleep-disordered breathing, such as obstructive sleep apnea.
Publisher: Elsevier BV
Date: 05-1992
DOI: 10.1016/0306-4522(92)90349-7
Abstract: Intra-axonal recordings were made from 24 afferent fibres of the superior laryngeal nerve in and around the nucleus tractus solitarius, in 26 pentobarbitone-anaesthetized cats. Conduction velocity ranged from 15 to 38 m/s. Four afferents were injected with horseradish peroxidase. They showed dense terminal arborization in the region of the ventral and ventrolateral subnuclei of the nucleus tractus solitarius, both rostral and caudal to the obex. Six other intra-axonal recordings were thought to originate from axons of neurons postsynaptic to superior laryngeal afferents one of these was injected with horseradish peroxidase and showed a similar arborization pattern to that of the afferent axons. In the same region, intracellular recordings were made from 124 neurons which responded to superior laryngeal nerve stimulation with excitatory postsynaptic potentials (mean latency 2.7 +/- 1.0 ms). Ninety-nine of these neurons were thought to receive a monosynaptic input. The stimulation threshold evoking these responses was similar to that which inhibited phrenic nerve discharge. Eleven of the monosynaptically excited neurons were injected with horseradish peroxidase. They had fusiform or stellate somata and simple dendritic trees, radiating mainly in the transverse plane. In one experiment, in which both a superior laryngeal nerve afferent fibre and a neuron were labelled, afferent terminal varicosities were found in close apposition with the postsynaptic membrane of the injected neuron. Four of 14 (29%) tested neurons could be antidromically activated from the C3 spinal segment. The stimulus thresholds and onset latencies of the responses of superior laryngeal nerve afferents and medullary neurons to stimulation of the superior laryngeal nerve are consistent with their involvement in the reflex inhibition of respiratory neurons evoked by superior laryngeal nerve stimulation.
Publisher: Elsevier BV
Date: 02-2018
DOI: 10.1016/J.NEULET.2018.11.024
Abstract: The airway vagal preganglionic neurons (AVPNs) in the external formation of the nucleus ambiguus (eNA) can be separated into inspiratory-activated AVPNs (IA-AVPNs) and inspiratory-inhibited AVPNs (II-AVPNs). IA-AVPNs are activated by excitatory presynaptic inputs during inspiratory bursts, but the composition and the roles of these excitatory inputs still remain obscure. II-AVPNs are inhibited by inhibitory presynaptic inputs but whether these inhibitory inputs are regulated by excitatory inputs is also unclear. In the current study, AVPNs were retrogradely fluorescent labeled. The IA-AVPNs were discriminated from II-AVPNs by their different synaptic inputs during inspiratory bursts. The excitatory inputs to IA-AVPNs and the presynaptic regulation of II-AVPNs were examined by whole-cell patch cl ing. Topical application of 6-Cyano-7-nitroquinoxaline-2,3-dione (CNQX) to the recorded IA-AVPNs almost abolished the tonic EPSCs during inspiratory intervals, inhibited the phasic excitatory currents during inspiratory bursts and attenuated the phasic inspiratory inward currents (PIICs) driven by central inspiratory activity. Blockade of α4β2 and α7 nicotinic acetylcholine receptors (nAChRs) respectively inhibited PIICs in some IA-AVPNs. Carbenoxolone, a gap junction uncoupler, partly inhibited the PIICs of IA-AVPNs. Focal application of CNQX to the II-AVPNs significantly inhibited the frequency, peak litude and area of the phasic inspiratory outward currents (PIOCs). These findings demonstrated that glutamatergic non-NMDA receptors played a predominant role in the excitatory drive to the IA-AVPNs, and that α4β2, α7 nAChRs and gap junctions were also rhythmically activated by central inspiratory activity. Additionally, glycinergic neurons making inhibitory inputs to the II-AVPNs were pre-synaptically facilitated by excitatory glutamatergic synaptic inputs.
Publisher: Elsevier BV
Date: 2020
Publisher: Wiley
Date: 22-06-2004
DOI: 10.1002/SYN.20044
Abstract: N-ethylmaleimide (NEM) has been used extensively in biochemical assays as an inhibitor of the NEM sensitive fusion protein (NSF). However, examination of the effect of NEM on transmitter release in more physiologically relevant preparations has proved inconclusive. In the present study, we have examined the effect of low concentrations of NEM on synaptic transmission in intact nerve-muscle preparations from toads (Bufo marinus). Under conditions of low transmitter release probability (0.3 mM calcium, 1 mM magnesium), treatment with NEM (10 microM) caused a significant increase in the litude of stimulus-evoked endplate potentials (EPPs) and a significant increase in the frequency of spontaneously occurring miniature EPPS (MEPPS) without affecting the litude of MEPPs. When the calcium concentration in the bath was raised to 4 mM, 10 microM NEM had no effect on EPP litude. Under these conditions, NEM treatment reduced paired pulse facilitation and increased depression during stimulus trains. Treatment with NEM also resulted in a significant decrease in the synaptic delay. The effects of NEM on transmitter release in the present study were not due to inactivation of G-proteins. The results of the present study show a calcium-dependent facilitation of stimulus-evoked transmitter release by NEM. These results are discussed in terms of the possible sites of NEM action leading to the observed changes in transmitter release.
Publisher: Elsevier BV
Date: 05-1989
DOI: 10.1016/0006-8993(89)90530-1
Abstract: Synaptic responses evoked in phrenic motoneurones (PMNs) by stimulation of the superior laryngeal nerve (SLN) were analysed in anaesthetised cats. Stimulation of the SLN was followed by inhibition of ipsilateral phrenic nerve discharge with the latency of 9.5 +/- 2.3 ms (mean +/- S.D.) and hyperpolarizations of ipsilateral PMN membrane potentials (latency, 8.4 +/- 2.1 ms) which were observed after stimuli applied both during inspiration and expiration. During the injection of Cl ions, the hyperpolarizations were either reversed or flattened in all 28 tested PMNs, thus indicating a direct inhibition. The possibility that the inhibitory postsynaptic potentials are produced by segmental respiratory interneurones is discussed.
Publisher: Elsevier BV
Date: 08-1994
DOI: 10.1016/0304-3940(94)90065-5
Abstract: Short-latency excitatory postsynaptic potentials (EPSPs), evoked by electrical stimulation lateral to the hypoglossal motor nucleus, were recorded from rat hypoglossal motoneurons (HMs) in brainstem slices. EPSPs were markedly suppressed or abolished by kynurenic acid (1 mM), showing that they were glutamatergic. The adenosine receptor agonist 2-chloro-N6-cyclopentyladenosine (CCPA, 100 nM) reduced EPSP litude to 42% of control, while the agonist 2-chloroadenosine (2-CA, 0.5-50 microM) caused a dose-dependent reduction of the EPSP. The adenosine receptor antagonist 8-cyclopentyl-1, 3-dipropylxanthine (DPCPX, 0.1-1 microM) increased the EPSP litude to 124% of control, and blocked EPSP reduction by CCPA or 2-CA. CCPA, 2-CA and DPCPX did not significantly alter HM input resistance or membrane potential. These data indicate that excitatory glutamatergic inputs to rat HMs are modulated by adenosine A1 receptors, most probably at a presynaptic site. This modulation may be especially significant in hypoxic responses of HMs.
Publisher: Hindawi Limited
Date: 2016
DOI: 10.1155/2016/3423267
Abstract: Motoneurons develop extensive dendritic trees for receiving excitatory and inhibitory synaptic inputs to perform a variety of complex motor tasks. At birth, the somatodendritic domains of mouse hypoglossal and lumbar motoneurons have dense filopodia and spines. Consistent with Vaughn’s synaptotropic hypothesis, we propose a developmental unified-hybrid model implicating filopodia in motoneuron spinogenesis/synaptogenesis and dendritic growth and branching critical for circuit formation and synaptic plasticity at embryonic renatal/neonatal period. Filopodia density decreases and spine density initially increases until postnatal day 15 (P15) and then decreases by P30. Spine distribution shifts towards the distal dendrites, and spines become shorter (stubby), coinciding with decreases in frequency and increases in litude of excitatory postsynaptic currents with maturation. In transgenic mice, either overexpressing the mutated human Cu/Zn-superoxide dismutase ( hSOD 1 G 93 A ) gene or deficient in GABAergic/glycinergic synaptic transmission (gephyrin, GAD-67, or VGAT gene knockout), hypoglossal motoneurons develop excitatory glutamatergic synaptic hyperactivity. Functional synaptic hyperactivity is associated with increased dendritic growth, branching, and increased spine and filopodia density, involving actin-based cytoskeletal and structural remodelling. Energy-dependent ionic pumps that maintain intracellular sodium/calcium homeostasis are chronically challenged by activity and selectively overwhelmed by hyperactivity which eventually causes sustained membrane depolarization leading to excitotoxicity, activating microglia to phagocytose degenerating neurons under neuropathological conditions.
Publisher: Ovid Technologies (Wolters Kluwer Health)
Date: 06-2004
DOI: 10.1097/01.WNR.0000127462.15985.DC
Abstract: Developmental changes in expression of two-pore domain K+ channels, TASK-1 and TREK-1, were investigated in the juvenile (postnatal day 13 P13) and adult (P105) rat brain stem and cerebellum using immunohistochemistry. In the juvenile, extensive TASK-1-like immunoreactivity (TASK-1-LIR) was seen among glial cells in the white matter (e.g., radial glia), which showed marked reduction in the adult. In contrast, TASK-1-LIR in neurons including cerebellar Purkinje and granule cells, hypoglossal and facial motoneurons, and ventrolateral medulla neurons was increased in the adult. TASK-1-LIR in neuroglia surrounding peripheral axons of cranial nerves was persistent. TREK-1-LIR was similar between ages, although TREK-1-LIR was neuronal and present only in juvenile cerebellar external germinal layer. Present results suggest roles for TASK-1 and K+ homeostasis in neuro-glial interaction, neurogenesis, differentiation, migration, axon guidance, synaptogenesis and myelination.
Publisher: Frontiers Media SA
Date: 2013
Publisher: American Physiological Society
Date: 10-2012
DOI: 10.1152/JAPPLPHYSIOL.00699.2011
Abstract: In brain stem slices from neonatal ( postnatal days 0–4) CD-1 mice, muscarinic ACh receptors (MAChRs) increased rhythmic inspiratory-related and tonic hypoglossal nerve discharge and depolarized single hypoglossal motoneurons (HMs) via an inward current without changing input resistance. These responses were blocked by the MAChR antagonist 1,1-dimethyl-4-diphenylacetoxypiperidinium iodide (4-DAMP 100 nM). MAChRs shifted voltage-dependent activation of the hyperpolarization-activated cation current to more positive levels. MAChRs increased the HM repetitive firing rate and decreased rheobase, with both effects being blocked by 4-DAMP. Muscarinic agonists reduced the afterhyperpolarization of single action potentials (APs), suggesting that small-conductance Ca 2+ -dependent K + current inhibition increased the HM firing rate. Muscarinic agonists also reduced the AP litude and slowed its time course, suggesting that MAChRs inhibited voltage-gated Na + channels. To compare muscarinic excitation of single HMs to muscarinic excitatory effects on motor output in thicker brain stem slices requiring higher extracellular K + for rhythmic activity, we tested the effects of muscarinic agonists on single HM excitability in high-K + artificial cerebrospinal fluid (aCSF). In high-K + aCSF, muscarinic agonists still depolarized HMs and altered AP size and shape, as in standard aCSF, but did not increase the steady-state firing rate, decrease afterhyperpolarization, or alter threshold potential. These results indicate that the basic cellular response of HMs to muscarinic receptors is excitatory, via a number of distinct mechanisms, and that this excitatory response will be largely preserved in rhythmically active brain stem slices.
Publisher: Frontiers Media SA
Date: 05-12-2017
Publisher: Frontiers Media SA
Date: 13-08-2019
Publisher: The Endocrine Society
Date: 23-05-2012
DOI: 10.1210/EN.2011-2171
Abstract: GH deficiency has been found in subjects with amyotrophic lateral sclerosis (ALS). Disrupted endocrine function could contribute to the progressive muscle loss and hypermetabolism seen in ALS. It is not possible to study all the elements of the GH-IGF-I axis in ALS patients. Consequently, it remains unclear whether dysfunctional GH secretion contributes to disease pathogenesis and why GH and IGF-I directed treatment strategies are ineffective in human ALS. The hSOD1G93A transgenic mouse model is useful for the detailed investigation of the pathogenesis of ALS. We report that symptomatic male hSOD1G93A transgenic mice exhibit a deficiency in GH secretion similar to that seen in human ALS. Further characterization of the GH-IGF-I axis in hSOD1G93A mice reveals central and peripheral abnormalities that are not found in wild-type age-matched controls. Specifically, we observe aberrant endogenous pulsatile GH secretion, reduced pituitary GH content, and decreased circulating levels of IGF-I, indicating global GH deficiency in hSOD1G93A mice. Furthermore, a reduction in the expression of the IGF-I receptor α-subunit in skeletal muscle and lumbar spinal cords of hSOD1G93A mice suggests impaired IGF-I signaling within these tissues. This is the first account of disrupted GH secretion in a transgenic mouse model of ALS. These observations are essential for the development of effective GH and IGF-I targeted therapies in ALS.
Publisher: Springer Science and Business Media LLC
Date: 17-10-2015
Publisher: Ovid Technologies (Wolters Kluwer Health)
Date: 12-1994
DOI: 10.1097/00001756-199412300-00010
Abstract: Intracellular pH (pHi) and membrane potential (Em) were measured in vivo in expiratory neurones and glial cells in the medulla of anaesthetized cats using double-barrelled H(+)-sensitive microelectrodes. In glial cells, stimulation of spinal pathways evoked a depolarization of up to 12 mV litude and an increase of pHi (7.25 +/- 0.15) by maximally 0.1 pH unit. IN expiratory neurones, pHi (7.15 +/- 0.18) fell by up to 0.2 pH unit during inspiratory inhibition. In axons of expiratory neurones, pHi remained unaffected during rhythmic action potential discharges. We suggest that the glial alkalinization is due to activation of Na+/HCO3- cotransport, whereas the neuronal acidification is caused by efflux of HCO3- via receptor-coupled anion channels.
Publisher: Wiley
Date: 30-09-2019
DOI: 10.1002/AR.24255
Abstract: The motor neuron (MN) soma surface area is correlated with motor unit type. Larger MNs innervate fast fatigue-intermediate (FInt) or fast-fatiguable (FF) muscle fibers in type FInt and FF motor units, respectively. Smaller MNs innervate slow-twitch fatigue-resistant (S) or fast fatigue-resistant (FR) muscle fibers in type S and FR motor units, respectively. In amyotrophic lateral sclerosis (ALS), FInt and FF motor units are more vulnerable, with denervation and MN death occurring for these units before the more resilient S and FR units. Abnormal MN dendritic arbors have been observed in ALS in humans and rodent models. We used a Golgi-Cox impregnation protocol to examine soma size-dependent changes in the dendritic morphology of lumbar MNs in SOD1
Publisher: Oxford University Press (OUP)
Date: 22-12-2017
DOI: 10.1093/HMG/DDW371
Abstract: Ataxia-telangiectasia (A-T), an autosomal recessive disease caused by mutations in the ATM gene is characterised by cerebellar atrophy and progressive neurodegeneration which has been poorly recapitulated in Atm mutant mice. Consequently, pathways leading to neurodegeneration in A-T are poorly understood. We describe here the generation of an Atm knockout rat model that does not display cerebellar atrophy but instead paralysis and spinal cord atrophy, reminiscent of that seen in older patients and milder forms of the disorder. Loss of Atm in neurons and glia leads to accumulation of cytosolic DNA, increased cytokine production and constitutive activation of microglia consistent with a neuroinflammatory phenotype. Rats lacking ATM had significant loss of motor neurons and microgliosis in the spinal cord, consistent with onset of paralysis. Since short term treatment with steroids has been shown to improve the neurological signs in A-T patients we determined if that was also the case for Atm-deficient rats. Betamethasone treatment extended the lifespan of Atm knockout rats, prevented microglial activation and significantly decreased neuroinflammatory changes and motor neuron loss. These results point to unrepaired damage to DNA leading to significant levels of cytosolic DNA in Atm-deficient neurons and microglia and as a consequence activation of the cGAS-STING pathway and cytokine production. This in turn would increase the inflammatory microenvironment leading to dysfunction and death of neurons. Thus the rat model represents a suitable one for studying neurodegeneration in A-T and adds support for the use of anti-inflammatory drugs for the treatment of neurodegeneration in A-T patients.
Publisher: Public Library of Science (PLoS)
Date: 15-02-2013
Publisher: American Physiological Society
Date: 04-2019
Abstract: The effect of capsaicin on glycinergic synaptic transmission to juvenile rat hypoglossal motor neurons in acute brainstem slices was evaluated in the presence of TTX. Capsaicin caused a robust decrease in miniature IPSC frequency, litude, and half-width, showing that this effect is independent of action potential generation. In the presence of capsazepine, a classic TRPV1 antagonist, capsaicin was still able to reduce spontaneous inhibitory postsynaptic current (IPSC) litude and frequency. We further investigated whether the effect of capsaicin on glycinergic transmission to hypoglossal motor neurons is pre- or postsynaptic in nature by recording pairs of evoked IPSCs. Interestingly, capsaicin also reduced evoked IPSC litude without affecting paired-pulse ratio, indicating a postsynaptic mechanism of action. Significant reduction was also observed in evoked IPSC half-width, rise time, and decay tau. We also show that capsaicin does not have any effect on either transient (It) or sustained (Is) potassium currents. Finally, we also show that the hyperpolarization-activated cationic current (Ih) also remains unchanged after capsaicin application. NEW & NOTEWORTHY Capsaicin reduces the litude of quantal and evoked glycinergic inhibitory neurotransmission to brainstem motor neurons without altering activity-dependent transmitter release. This effect of capsaicin is not due to activation of TRPV1 receptors, as it is not blocked by capsazepine, a TRPV1 receptor antagonist. Capsaicin does not alter voltage-dependent potassium current or the hyperpolarization-activated cationic current in brainstem motor neurons.
Publisher: Springer Science and Business Media LLC
Date: 29-11-2016
DOI: 10.1038/SREP37968
Abstract: Layer V pyramidal neurons (LVPNs) within the motor cortex integrate sensory cues and co-ordinate voluntary control of motor output. In amyotrophic lateral sclerosis (ALS) LVPNs and spinal motor neurons degenerate. The pathogenesis of neural degeneration is unknown in ALS 10% of cases have a genetic cause, whereas 90% are sporadic, with most of the latter showing TDP-43 inclusions. Clinical and experimental evidence implicate excitotoxicity as a prime aetiological candidate. Using patch cl and dye-filling techniques in brain slices, combined with high-resolution confocal microscopy, we report increased excitatory synaptic inputs and dendritic spine densities in early presymptomatic mice carrying a TDP-43 Q331K mutation. These findings demonstrate substantive alterations in the motor cortex neural network, long before an overt degenerative phenotype has been reported. We conclude that increased excitatory neurotransmission is a common pathophysiology amongst differing genetic cases of ALS and may be of relevance to the 95% of sporadic ALS cases that exhibit TDP-43 inclusions.
Publisher: Wiley
Date: 19-04-2013
DOI: 10.1111/JVP.12055
Abstract: Alfaxalone (3α-hydroxy-5α-pregnane-11, 20-dione) is a neuroactive steroid with anaesthetic properties and a wide margin of safety. The pharmacokinetic properties of alfaxalone administered intravenously and intraperitoneally in rats (n = 28) were investigated. Mean t(1/2elim) for 2 and 5 mg/kg i.v. was 16.2 and 17.6 min, respectively, but could not be estimated for IP dosing, due to sustained plasma levels for up to 60 min after injection. Clp for i.v. injection was calculated at 57.8 ± 23.6 and 54.3 ± 6.8 mL/min/kg, which were 24.5% and 23% of cardiac output, respectively. The observed C(max) was 3.0 mg/L for IP administration, and 2.2 ± 0.9 and 5.2 ± 1.3 mg/L for 2 and 5 mg/kg i.v. administration, respectively. AUC(0-60) was 96.2 min.mg/L for IP dosing. The relative bioavailability for IP dosing was 26% and 28% compared to i.v. dosing. Differences in t(1/2elim) and Cl(p) from previous pharmacokinetic studies in rats are likely due to variations in alfaxalone formulation rather than sex differences. Alfaxan® given IP caused sustained levels of alfaxalone, no apnoea and longer sleep times than i.v. dosing, although immobilization was not induced in 30% of rats given Alfaxan® IP. A pharmacodynamic study of the effects of combining IP injection of Alfaxan® with other premedication agents is worthwhile, to determine whether improved anaesthesia induction could ultimately provide an alternative anaesthetic regimen for rats.
Publisher: American Physiological Society
Date: 09-2013
Abstract: Riluzole is the sole treatment for amyotrophic lateral sclerosis (ALS), but its therapeutically relevant actions on motor neurons are not well defined. Whole cell patch-cl recordings were made from hypoglossal motor neurons (HMs, n = 25) in brain stem slices from 10- to 23-day-old rats anesthetized with pentobarbital sodium to investigate the hypothesis that riluzole inhibits HMs by multiple mechanisms. Riluzole (20 μM) hyperpolarized HMs by decreasing an inward current, inhibited voltage-gated persistent Na + and Ca 2+ currents activated by slow voltage r s, and negatively shifted activation of the hyperpolarization-activated cationic current ( I H ). Repetitive firing of HMs was strongly inhibited by riluzole, which also increased action potential threshold voltage and rheobase and decreased litude and maximum rise slope but did not alter the maximal afterhyperpolarization litude or decay time constant. HM rheobase was inversely correlated with persistent Na + current density. Glutamatergic synaptic transmission was inhibited by riluzole by both pre- and postsynaptic effects. Riluzole decreased activity-dependent glutamate release, as shown by decreased litude of evoked and spontaneous excitatory postsynaptic currents (EPSCs), decreased paired-pulse ratio, and decreased spontaneous, but not miniature, EPSC frequency. However, riluzole also decreased miniature EPSC litude and the inward current evoked by local application of glutamate onto HMs, suggesting a reduction of postsynaptic glutamate receptor sensitivity. Riluzole thus has a marked inhibitory effect on HM activity by membrane hyperpolarization, decreasing firing and inhibiting glutamatergic excitation by both pre- and postsynaptic mechanisms. These results broaden the range of mechanisms controlling motor neuron inhibition by riluzole and are relevant to researchers and clinicians interested in understanding ALS pathogenesis and treatment.
No related grants have been discovered for Mark Bellingham.