Somatic Motor Nerve Research Articles

Автономная диабетическая нейропатия

Diabetes mellitus (DM) is widespread in Russia, which has caused an increase in the frequency of complications in this nosology, including diabetic autonomic neuropathy (DАN). DАN determines an increase in mortality and a decrease in life expectancy. The purpose — to study the clinical manifestations of diabetic autonomic neuropathy. Results. In DAN, a lesion of various organ systems is observed. Manifestations of DAN are usually preceded by the development of typical signs of distal sensorimotor polyneuropathy. In some patients, manifestations of damage to the autonomic nervous system may prevail over damage to somatic sensory and motor nerve fibers. It should be borne in mind that in more than half of cases, DAN is accompanied by a fatal outcome with a duration of DM of more than 10 years. Cardiovascular autonomic neuropathy (CAN), which determines life expectancy in DM, is most often studied in the modern Russian and foreign medical science. The studies of other DAN manifestations are presented with much less frequency. DAN is accompanied by damage to all parts of the autonomic nervous system and determines an increase in mortality in patients with DM, which requires early detection of signs of diabetic autonomic neuropathy. Conclusion. Thus, DAN can cause an increase in mortality, a persistent decrease in working capacity in patients with DM, which requires general practitioners to be aware of the features of clinical signs of this nosology.

Bilateral Anatomical Variation in the Formation of Trunks of the Brachial Plexus - A Case Report

AbstractThis study presents a bilateral variation in the formation of trunks of brachial plexus in a male cadaver. The right brachial plexus was composed of six roots (C4-T1) and the left brachial plexus of five roots (C5-T1). Both formed four trunks thus changing the contributions of the anterior divisions of the cervical nerves involved in the formation of the cords and the five main somatic motor nerves for the upper limb. There are very few case reports in the scientific literature on this topic; thus making the present study very relevant.

Distribution of the creatine transporter throughout the human brain reveals a spectrum of creatine transporter immunoreactivity

ABSTRACTCreatine is a molecule that supports energy metabolism in cells. It is carried across the plasma membrane by the creatine transporter. There has been recent interest in creatine for its neuroprotective effects in neurodegenerative diseases and its potential as a therapeutic agent. This study represents the first systematic investigation of the distribution of the creatine transporter in the human brain. We have used immunohistochemical techniques to map out its location and the intensity of staining. The transporter was found to be strongly expressed, especially in the large projection neurons of the brain and spinal cord. These include the pyramidal neurons in the cerebral cortex, Purkinje cells in the cerebellar cortex, and motor neurons of the somatic motor and visceromotor cranial nerve nuclei and the ventral horn of the spinal cord. Many other neurons in the brain also had some degree of creatine transporter immunoreactivity. By contrast, the medium spiny neurons of the striatum and the catecholaminergic neurons of the substantia nigra and locus coeruleus, which are implicated in neurodegenerative diseases, showed a very low to almost absent level of immunoreactivity for the transporter. We propose that the distribution may reflect the energy consumption by different cell types and that the extent of creatine transporter expression is proportional to the cell's energy requirements. Furthermore, the distribution indicates that supplemented creatine would be widely taken up by brain cells, although possibly less by those cells that degenerate in Huntington's and Parkinson's diseases. J. Comp. Neurol. 523:699–725, 2015. © 2014 Wiley Periodicals, Inc.

Sensory Guillain-Barré syndrome: A case report.

A 58-year-old female exhibited the onset of symmetrical sensory abnormalities of the face and extremities. The neurological examination revealed normal muscle strength with abated or absent tendon reflexes. The patient experienced symmetrical glove- and stocking-type pinprick sensations in the distal extremities and a loss of temperature sensation, but had normal proprioception and vibration senses and joint topesthesia. The lumbar puncture showed protein cell separation at the fifth week after the onset of symptoms. At the same time-point, the electrophysiological examination showed demyelination changes involving the trigeminal nerve and the somatic motor nerve. Needle electromyography revealed normal results. The clinical symptoms ceased progression at the fourth week after symptom onset, and began to improve from the sixth. This case was considered to be sensory Guillain-Barré syndrome, which was characterized by its cranial nerve involvement.

Distribution of the creatine transporter throughout the human brain reveals a spectrum of creatine transporter immunoreactivity.

Creatine is a molecule that supports energy metabolism in cells. It is carried across the plasma membrane by the creatine transporter. There has been recent interest in creatine for its neuroprotective effects in neurodegenerative diseases and its potential as a therapeutic agent. This study represents the first systematic investigation of the distribution of the creatine transporter in the human brain. We have used immunohistochemical techniques to map out its location and the intensity of staining. The transporter was found to be strongly expressed, especially in the large projection neurons of the brain and spinal cord. These include the pyramidal neurons in the cerebral cortex, Purkinje cells in the cerebellar cortex, and motor neurons of the somatic motor and visceromotor cranial nerve nuclei and the ventral horn of the spinal cord. Many other neurons in the brain also had some degree of creatine transporter immunoreactivity. By contrast, the medium spiny neurons of the striatum and the catecholaminergic neurons of the substantia nigra and locus coeruleus, which are implicated in neurodegenerative diseases, showed a very low to almost absent level of immunoreactivity for the transporter. We propose that the distribution may reflect the energy consumption by different cell types and that the extent of creatine transporter expression is proportional to the cell's energy requirements. Furthermore, the distribution indicates that supplemented creatine would be widely taken up by brain cells, although possibly less by those cells that degenerate in Huntington's and Parkinson's diseases.

Immunostaining, Dehydration, and Clearing of Mouse Embryos for Ultramicroscopy

This protocol describes the preparation of mouse embryos for ultramicroscopy (UM), a powerful imaging technique that achieves precise and accurate three-dimensional (3D) reconstructions of intact macroscopic specimens with micrometer resolution. In UM, a specimen in the size range of ∼1-15 mm is illuminated perpendicular to the observation pathway by two thin counterpropagating sheets of laser light. In combination with fluorescein isothiocyanate (FITC) immunostaining, UM allows visualization of somatic motor and sensorial nerve fibers in whole mouse embryos. Even the fine branches of the sensomotoric fibers can be visualized over a distance of up to several millimeters. In this protocol, mouse embryos are fixed and immunostained in preparation for UM. Because UM requires the excitation light sheet to travel throughout the entire horizontal width of the specimen, specimens usually have to be rendered transparent before microscope inspection. Here, the embryos are dehydrated in ethanol and then cleared in a solution of benzyl alcohol and benzyl benzoate.

Differential Neuropathies in Hyperglycemic and Hypoglycemic Diabetic Rats

We investigated the effects of hyperglycemia and hypoglycemia on development of peripheral neuropathy in somatic motor and sensory nerves in type 1 diabetic BB/Wor rats. The animals were maintained in a hyper- or hypoglycemic state by treatment with insulin for 3 months. Nondiabetic siblings served as controls. Qualitative analysis of the gastrocnemius and sural nerves by light and electron microscopy revealed signs of Wallerian-type axonal degeneration and regeneration of large myelinated fibers in the hypoglycemic but not the hyperglycemic animals. Degeneration was more common in the gastrocnemius nerve than in the sural nerve. In hypoglycemic rats, myelinated fibers in both the gastrocnemius and sural nerves had significantly shorter internodes and smaller diameters. The decreased fiber diameter was related (r = -0.9) to the duration of severe hypoglycemia (</=2.5 mmol/L). Myelinated fiber occupancy was also decreased without any significant changes in fiber counts in both the gastrocnemius and sural nerves. In hyperglycemic rats, myelinated fibers in the sural nerve but not the gastrocnemius nerve had smaller diameters compared with controls. We conclude that hypoglycemia has a more severe impact on somatic motor nerves than on somatic sensory nerves, whereas hyperglycemia affects only somatic sensory nerves.

Peripheral nerve functions may deteriorate parallel to the progression of microangiopathy in diabetic patients

Our aim was to obtain information to help in the early detection of impaired nerve function and to assess the severity of diabetic symmetric polyneuropathy (DPN). Various somatic and autonomic nerve functions in 40 diabetics and 20 age-matched healthy volunteers were evaluated using six objective examinations: nerve conduction study, quantitative vibratory perception threshold, heart rate variability, Valsalva test, head-up tilt and quantitative sudomotor axonal reflex test (QSART). The diabetics were divided into three groups according to the severity of their microangiopathy. The nerve function data and level of impairment were compared between a healthy control and three diabetic groups. The relationships between nerve function data and clinical background were also examined using multivariate analysis. Results were as follows: (1) all nerve dysfunctions seemed to develop parallel to the progression of microangiopathy, (2) reduced nerve conduction velocity and elevated vibratory perception thresholds in the feet might be early detectable signs of DPN, (3) vasomotor and sudomotor sympathetic functions and cardiovagal functions seemed to deteriorate with the appearance of microangiopathy, (4) lowered compound muscle action potential seemed to appear at the advanced microangiopathic condition, (5) hypohydrosis was closely related to diabetic foot ulcers. In conclusion, nerve dysfunction in diabetics might generally progress with microangiopathy from somatic sensory nerve dysfunction to autonomic nerve dysfunction and then to somatic motor nerve dysfunction. Sympathetic sudomotor dysfunction might be a sensitive predictor of diabetic foot ulcer.

An electrophysiological study on the artificial somato-autonomic pathway for inducing voiding

To investigate the possibility of regeneration of somatic motor nerve to replace splanchnic nerve and the electrophysiologic characters of the regenerated nerve. An artificial somato-autonomic reflex pathway was established by intradural microanastomosis of L(4) ventral root (VR) to L(6)VR at the left side in 12 male Wistar rats. Then the L(4)VR proximal to the anastomosis was stimulated by silver electrode and the evoked potentials were recorded on the distal end to the anastomosis, pelvic nerve and postganlionic fibers of the major pelvic ganglia (MPG). Cystometrography was used to record the intravesical pressure. Hexamethonium, a cholinergic ganglion blocker, was given directly on the pelvic ganglion so as to observe the change of the intravesical pressure evoked by stimulation of the nerves. Another 12 rats were used as controls. (1) In the experimental group, stimulation of the L(4)VR proximal end to the anastomosis evoked potentials on the distal end, the pelvic nerve, and the postganglionic fibers of the MPG, and induced bladder contraction. Stimulation of the contralateral sciatic nerve failed to evoke change of intravesical pressure. In the control group stimulation of the L(4)VR or sciatic nerve failed to evoke potentials on the postganglionic fibers of pelvic nerve and change of intravesical pressure. (2) Stimulation of the ipsilateral sciatic nerve led to an increase of intravesical pressure. (3) After the use of hexamethonium stimulation of the ipsilateral sciatic nerve and proximal end of L(4)-L(6) anastomosis failed to evoke change of intravesical pressure. (4) The conduction velocity of the regenerated motor axons was 33.3 m/s +/- 6.9m/s, significantly higher than that of the control group (11.6 m/s +/- 1.6 m/s). Somatic motor axons can regenerate to the MPG and reinnervate the bladder and the impulses from the somatic motor neurons can initiate voiding.

Neurovascular Alignment in Adult Mouse Skeletal Muscles

Muscle blood flow increases with motor unit recruitment. The physical relationships between somatic motor nerves, which control muscle fiber contraction, and arterioles, which control microvascular perfusion, are unexplored. The authors tested the hypothesis that motor axons align with arterioles in adult skeletal muscle. Transgenic mice (C57BL/6 background, n = 5; 10 months of age) expressing yellow fluorescent protein in all motor nerves underwent vascular casting (Microfil). Excised epitrochlearis, gracilis, gluteus maximus, and spinotrapezius muscles were imaged at 380x and 760x and a computer-integrated tracing system (Neurolucida) was used to acquire 3-dimensional digital renderings of entire arteriolar and neural networks within each muscle. Arteriolar networks were typically approximately 3-fold longer than neural networks. Nerves coursed with arterioles until terminating at motor endplates. Across muscles, proximity analyses revealed that approximately 75% of total nerve length (9.8-48.8 mm) lay within 200 microm of the nearest arteriole (diameters of 15-60 microm). Somatic motor nerves and arterioles align closely within adult mammalian skeletal muscle. Understanding the signals governing neurovascular alignment may hold important clues for the advancement of tissue engineering and regeneration.

Editorial

Editorial

Cutaneous sympathetic motor rhythms during a fever-like response induced by prostaglandin E 1

Neuronal population discharges within the CNS and in somatic and sympathetic motor nerves often display oscillations. Peripheral oscillations may provide a window into central mechanisms, as they often show coherence with population activity of subsets of central neurones. The reduction in heat loss through the cutaneous circulation during fever may be mediated via sympathetic premotor neurones not utilised during normal temperature regulation. Consequently, here we assessed, in anaesthetised rats, whether the frequency signature of population sympathetic discharge observed in neurones innervating the tail (thermoregulatory) circulation changed during a fever-like response induced by intracerebroventricular injection of prostaglandin E 1. We found that when core temperature was raised to 38.8–40.5°C sympathetic activity was abolished. Following administration of prostaglandin (400 ng or 1 μg per rat), activity was restored to levels seen prior to heating (154±53.5%; n=10). Injection of vehicle had no effect ( n=7). Prior to heating when most animals were in central apnoea (14/18) two peaks were observed in autospectra of sympathetic activity: one at 0.68–0.93 Hz (T-peak) and another at the frequency of ventilation (2 Hz). Central respiratory drive was recruited during hyperthermia where it was 1:2 locked to the frequency of ventilation and following prostaglandin administration, an additional peak in sympathetic autospectra was seen at this frequency. Time-evolving spectra indicated that this peak resulted from the dynamic locking of the ‘T-peak’ to central respiratory drive. Our data show that during a fever-like response the dominant oscillations in sympathetic activity controlling a thermoregulatory circulation and their dynamic coupling to respiratory-related inputs are similar to those seen under normal conditions. Therefore, during this fever-like response, the neural substrate(s) underlying the oscillations is not reconfigured and remains capable of sculpturing the pattern of sympathetic neuronal discharge that may be regulated by several descending pathways.

Synchronization of somato-sympathetic outflows during exercise: role for a spinal rhythm generator.

The synchronization of somatic motor outflow and sympathetic efferent activity during exercise has important functional consequences. For example, a certain degree of sympathetic vasoconstriction to active skeletal muscle vascular beds may be required to prevent an excessive decrease in vascular resistance during intense dynamic exercise involving a large skeletal muscle mass. Moreover, the redistribution of blood flow away from non-exercising vascular beds to contracting skeletal muscles is necessary to meet their increased metabolic demands. Historically, the nervous system has been separated arbitrarily into a volitional (somatic) and a reflex (autonomic) control system that does not necessitate volitional input (Koizumi & Brooks, 1972). However, during exercise this partition of the somatic and autonomic control systems is replaced by an overlapping control system that influences both somatic and sympathetic outflows in a parallel fashion. Previous work has suggested that the integration and co-ordination of the somatic and sympathetic motor responses during exercise were mediated by a supraspinal neural mechanism referred to as central command (Waldrop et al. 1996). However, there is an accumulating body of evidence, including papers on the role of the spinal cord in the regulation of sympathetic outflow (Weaver & Stein, 1989), suggesting that rhythmic neural activity destined for somatomotor and sympathetic outflows may be under the control of intrinsic neuronal circuitry in the spinal cord. In this issue of The Journal of Physiology, the paper by Chizh et al. (1998) demonstrates for the first time that some components of somatic motor and sympathetic neural outflows are synchronized by a spinal mechanism. The authors studied the relationship between sympathetic and somatic motor outflows from the thoraco-sacral spinal cord using an isolated trunk-hindquarters preparation of the adult mouse. They found that some of the activity in both somatic and sympathetic outflows was coupled. However, since there was also sympathetic neural activity that showed burst discharges in the absence of somatic motor activity, the spinal mechanism responsible for generating sympathetic burst activity was not the same mechanism that was responsible for somato-sympathetic coupling. Therefore, there remained spinal mechanisms that were capable of generating bursting activity independently of somatic and sympathetic activities. When NMDA was added to the perfusate to induce rhythmic somatic motor activity, a dose-dependent bursting pattern in sympathetic efferent and somatic motor nerves was evoked that was coupled at similar frequencies. Correlational analyses found a strong coherence between somatic motor outflow and sympathetic efferent discharge when triggered from peaks in somatic motor activity. However, this synchronization was not found when triggered from the peaks of sympathetic discharge. This finding led Chizh et al. (1998) to hypothesize that the generating mechanism responsible for somatic locomotor activity was the same mechanism responsible for coupling somatic outflow to sympathetic activity. These findings support the notion that the spinal cord possesses the necessary neuronal ‘machinery’ for generating and coupling sympathetic discharge with somatic motor outflow independently of supraspinal structures (i.e. the spinal rhythm generator represents a spinal central command). Important adjustments occur in the heart and blood vessels during exercise in order to provide the appropriate oxygen delivery to match the metabolic demand of contracting skeletal muscle. The coupling of somatic motor and sympathetic outflows is mediated by supraspinal neural mechanisms (supraspinal central command) in humans and animals (Fig. 1). In humans, intense intermittent static exercise produced a one-to-one synchronization of somatic motor activity with muscle sympathetic nerve activity that was purportedly directed to skeletal muscle vasculature (Victor et al. 1995). This synchronization persisted in the presence of both partial neuromuscular blockade (i.e. to increase central command) and after local anaesthetic block of the arm nerves to eliminate afferent feedback from skeletal muscle. Likewise, electrical and chemical stimulation of the hypothalamic locomotor region in decorticate cats after neuromuscular blockade evokes fictive locomotion that produces increases in arterial blood pressure (Waldrop et al. 1996) and synchronization of sympathetic nerve activity with hindlimb motor nerve activity (Hajduczok et al. 1991). And, as seen in humans, the synchronization of somatic motor activity and sympathetic discharge was unaffected by peripheral feedback mechanisms. These findings suggest that central command signals of supraspinal origin can regulate and co-ordinate sympathetic and somatomotor outflows during exercise. Figure 1 Synchronization of somatic motor outflow and sympathetic efferent activity during exercise is mediated by supraspinal and spinal mechanisms. The important work of Chizh et al. (1998) extends our understanding of the synchronization of somatic motor activity and sympathetic outflow by highlighting the role of a spinal somato-sympathetic generator (Fig. 1). Also, the coupling of somato-sympathetic neural outflows in the spinal cord may represent an exquisite mechanism to finely tune sympathetic discharge to the segmental level of somatic motor outflow during exercise. Finally, {fontsize the degree to which supraspinal central command signals influence the somato-sympathetic spinal generator needs to be studied to further our understanding of somato-sympathetic coupling during volitional exercise.

Pax-6 is involved in the specification of hindbrain motor neuron subtype.

Pax-6 is a member of the vertebrate Pax gene family, which is structurally related to the Drosophila pair-rule gene, paired. In mammals, Pax-6 is expressed in several discrete domains of the developing CNS and has been implicated in neural development, although its precise role remains elusive. We found a novel Small eye rat strain (rSey2) with phenotypes similar to mouse and rat Small eye. Analyses of the Pax-6 gene revealed one base (C) insertion in an exon encoding the region downstream of the paired box of the Pax-6 gene, resulting in generation of truncated protein due to the frame shift. To explore the roles of Pax-6 in neural development, we searched for abnormalities in the nervous system in rSey2 homozygous embryos. rSey2/rSey2 exhibited abnormal development of motor neurons in the hindbrain. The Islet-1-positive motor neurons were generated just ventral to the Pax-6-expressing domain both in the wild-type and mutant embryos. However, two somatic motor (SM) nerves, the abducent and hypoglossal nerves, were missing in homozygous embryos. By retrograde and anterograde labeling, we found no SM-type axonogenesis (ventrally growing) in the mutant postotic hindbrain, though branchiomotor and visceral motor (BM/VM)-type axons (dorsally growing) were observed within the neural tube. To discover whether the identity of these motor neuron subtypes was changed in the mutant, we examined expression of LIM homeobox genes, Islet-1, Islet-2 and Lim-3. At the postotic levels of the hindbrain, SM neurons expressed all the three LIM genes, whereas BM/VM-type neurons were marked by Islet-1 only. In the Pax-6 mutant hindbrain, Islet-2 expression was specifically missing, which resulted in the loss of the cells harboring the postotic hindbrain SM-type LIM code (Islet-1 + Islet-2 + Lim-3). Furthermore, we found that expression of Wnt-7b, which overlapped with Pax-6 in the ventrolateral domain of the neural tube, was also specifically missing in the mutant hindbrain, while it remained intact in the dorsal non-overlapping domain. These results strongly suggest that Pax-6 is involved in the specification of subtypes of hindbrain motor neurons, presumably through the regulation of Islet-2 and Wnt-7b expression.

Quantal Transmitter Release onto Different Patterns of Receptor Distributions at Somatic Neuromuscular Junctions

The effect of different distributions of acetylcholine receptors at the active zone of somatic motor-nerve terminals on the amplitude-frequency histogram of spontaneous excitatory currents due to the secretion of a quantum of transmitter has been analysed by numerically solving the diffusion and binding equations. The time course and amplitude of the spontaneous currents for different arrangements of receptors has been determined. Organization of receptors into stripes of different patterns beneath the active zone, which is known to occur, does not affect the amplitude or temporal characteristics of the spontaneous currents. Increases in the amount of transmitter in a quantum released onto the stripes does not introduce any subnit structure into the spontaneous currents. It is concluded that if subunits are detectable in measured amplitude–frequency histograms of spontaneous potentials they arise because of presynaptic rather than postsynaptic factors.

Synaptic organization of amphibian sympathetic ganglia

The synaptic organization of the amphibian sympathetic ganglia was studied, especially in the last two abdominal paravertebral ganglia of the frog. These ganglia appear to form a monosynaptic relay, not containing interneurons. They consist of two systems working in parallel: the principal neurons, by far the most numerous, and a small number of chromaffin (i.e., SIF) cells, usually arranged in clusters. Each principal neuron is innervated by a preganglionic branch forming a set of cholinergic synapses which exhibit classical ultrastucture. The only peculiarity is the presence of a subsynaptic apparatus in a variable percentage of synaptic complexes. Electro-physiological studies have demonstrated that synaptic transmission is due to ACh release and involves several postsynaptic potentials. Moreover, the principal neurons are of two types, B and C, whose preganglionic axons and their own axons have different conduction velocities. C neurons tend to be small in diameter, and B neurons are larger, but the size distribution of the two populations overlaps. More recently, it was demonstrated that these two neuronal systems have different immunocytochemical features. The C preganglionic fibers contain an LHRH-like peptide, which is responsible for late synaptic events. The B preganglionic fibers contain CGRP, whose role has not yet been established. The principal neurons all contain adrenaline, but neuropeptide Y is also present in C neurons and could be a second transmitter at peripheral junctions. SP-containing fibers also pass through the ganglia, but give rise to intraganglionic synapses only rarely, except in the celiac plexus. Galanin can coexist with neuropeptide Y in certain C neurons. Numerous principal neurons are immunoreactive for VIP. Chromaffin cells contain noradrenaline and metenkephalin, and some contain SP or LHRH; they are endocrine cells controlled by preganglionic fibers and can have a modulatory effect on principal neurons endowed with appropriate receptors. The accessibility of frog abdominal ganglia and the anatomical separation of B and C preganglionic fiber pathways provide interesting systems in which to carry out experimentation on the stability and specificity of synaptic contacts. After postganglionic axotomy, the majority of synapses disappear by disruption of synaptic contacts. There is a certain discrepancy between the recovery of synaptic transmission and the reappearance of morphologically identifiable synapses, suggesting that a certain amount of transmission is possible at contacts devoid of synaptic complexes. The selective deafferentation of B or C neurons showed that the subsynaptic apparati are mainly found at B neuron synapses. The course of reinnervation following selective deafferentation reveals the existence of different specificities at B and C synapses: C neurons are easily reinnervated by B preganglionic fibers, whereas C fibers appear fairly ineffective at reinnervating B neurons, even after a long interval. Attempts were made to reinnervate ganglionic neurons with somatic motor nerve fibers. Reinnervation was achieved only rarely, and it is concluded that the ganglionic synapses in the frog have a higher specificity and lower plasticity than in mammals. © 1996 Wiley-Liss, Inc.

Synaptic organization of amphibian sympathetic ganglia.

The synaptic organization of the amphibian sympathetic ganglia was studied, especially in the last two abdominal paravertebral ganglia of the frog. These ganglia appear to form a monosynaptic relay, not containing interneurons. They consist of two systems working in parallel: the principal neurons, by far the most numerous, and a small number of chromaffin (i.e., SIF) cells, usually arranged in clusters. Each principal neuron is innervated by a preganglionic branch forming a set of cholinergic synapses which exhibit classical ultrastructure. The only peculiarity is the presence of a subsynaptic apparatus in a variable percentage of synaptic complexes. Electrophysiological studies have demonstrated that synaptic transmission is due to ACh release and involves several postsynaptic potentials. Moreover, the principal neurons are of two types, B and C, whose preganglionic axons and their own axons have different conduction velocities. C neurons tend to be small in diameter, and B neurons are larger, but the size distribution of the two populations overlaps. More recently, it was demonstrated that these two neuronal systems have different immunocytochemical features. The C preganglionic fibers contain an LHRH-like peptide, which is responsible for late synaptic events. The B preganglionic fibers contain CGRP, whose role has not yet been established. The principal neurons all contain adrenaline, but neuropeptide Y is also present in C neurons and could be a second transmitter at peripheral junctions. SP-containing fibers also pass through the ganglia, but give rise to intraganglionic synapses only rarely, except in the celiac plexus. Galanin can coexist with neuropeptide Y in certain C neurons. Numerous principal neurons are immunoreactive for VIP. Chromaffin cells contain noradrenaline and metenkephalin, and some contain SP or LHRH; they are endocrine cells controlled by preganglionic fibers and can have a modulatory effect on principal neurons endowed with appropriate receptors. The accessibility of frog abdominal ganglia and the anatomical separation of B and C preganglionic fiber pathways provide interesting systems in which to carry out experimentation on the stability and specificity of synaptic contacts. After postganglionic axotomy, the majority of synapses disappear by disruption of synaptic contacts. There is a certain discrepancy between the recovery of synaptic transmission and the reappearance of morphologically identifiable synapses, suggesting that a certain amount of transmission is possible at contacts devoid of synaptic complexes. The selective deafferentation of B or C neurons showed that the subsynaptic apparati are mainly found at B neuron synapses. The course of reinnervation following selective deafferentation reveals the existence of different specificities at B and C synapses: C neurons are easily reinnervated by B preganglionic fibers, whereas C fibers appear fairly ineffective at reinnervating B neurons, even after a long interval. Attempts were made to reinnervate ganglionic neurons with somatic motor nerve fibers. Reinnervation was achieved only rarely, and it is concluded that the ganglionic synapses in the frog have a higher specificity and lower plasticity than in mammals.

Quantal transmitter release at somatic motor-nerve terminals: stochastic analysis of the subunit hypothesis

Here we analyze the problem of determining whether experimentally measured spontaneous miniature end-plate currents (MEPCs) indicate that quanta are composed of subunits. The properties of MEPCs at end plates with or without secondary clefts at the neuromuscular junction are investigated, using both stochastic and deterministic models of the action of a quantum of transmitter. It is shown that as the amount of transmitter in a quantum is increased above about 4000 acetylcholine (ACh) molecules there is a linear increase in the size of the MEPC. It is possible to then use amplitude-frequency histograms of such MEPCs to detect a subunit structure, as there is little potentiation effect above 4000 ACh molecules. Autocorrelation and power spectral analyses of such histograms establish that their subunit structure can be detected if the coefficient of variation of the subunit size is less than about 0.12 or, if electrical noise is added, about 0.1. Positive gradients relate the rise time and half-decay times of MEPCs to their amplitude, even in the absence of potentiating effects; these gradients are shallower at motor nerve terminals that possess secondary clefts. The effect of asynchronous release of subunits is also investigated. The criteria determined by this analysis for identifying a subunit composition in the quantum are applied to an amplitude-frequency histogram of MEPCs recorded from a small group of active zones at a visualized amphibian motor-nerve terminal. This did not provide evidence for a subunit structure.

Probabilistic secretion of quanta at somatic motor-nerve terminals : the fusion-pore model, quantal detection and autoinhibition

The probability of detecting first, second, and later quanta secreted at release sites of a motor-nerve terminal during the early release period following a nerve impulse has been addressed. The possibility that early quantal release autoinhibits later quantal release during this period has also been ascertained. In this investigation, a model for the secretion of a quantum at a release site is developed in which, following the influx and diffusion of calcium ions to a release site protein associated with synaptic vesicles, kappa steps of association of the ions with the protein then occur at rate alpha. The release site protein then undergoes a conformational change which may not go on to completion if calcium ions dissociate from the protein at rate gamma. If this process does reach completion then a fusion-pore between the vesicle and the presynaptic membrane is created; this happens at rate delta. Key assumptions of this fusion-pore model are that the quantal secretions from each site are independent of each other, and that there is a large number of vesicles, each with a small probability of secretion, so that the number of secretions is Poisson in nature. These assumptions allow analytical expressions to be obtained for predicting the times at which first, second and later quanta are secreted during the early release period following an impulse. To test the model, experiments were performed in which the times of first, second and later quantal releases were determined at discrete regions along the length of visualized motor-terminal branches in toad (Bufo marinus) muscles. Estimates of model rate constants and of kappa from the times for first quantal secretions failed to give satisfactory predictions of the observed times of later secretions. Therefore, either the model fails, or the procedure used for detecting later quantal events as a consequence of their being masked by earlier quantal events is inadequate. To solve this detection problem, a two-dimensional analysis of the spread of charge following the secretion of a quantum at a random site on the motor-terminal branch has been done. This allows determination of the probability that later quanta will be detected following secretion of earlier quanta. The detection model was then incorporated into the fusion-pore model to predict the times at which second and later quanta occur during the early release period, based on the estimates of the model parameters derived from the analysis of first quantal releases. Good estimates were now obtained for the observed times of second and later quantal releases, indicating that appropriate procedures must be adopted for adequate detection of quantal secretions. Furthermore, the experiments provide support for the fusion-pore model. It has been suggested that the binomial nature of quantal release from the entire motor-nerve terminal may be explained if early quantal release inhibits later quantal release during the early quantal release phase (M. R. Bennett & J. Robinson 1990, Proc. R. Soc. Lond. B 239, 329-358). Although the fusion-pore detection error model gave good predictions of the observed times of first, second and later quantal releases, these may be improved if a model for autoinhibition is included. In this model the first quantum was taken as giving rise to an inhibition of secretion that propagates to surrounding release sites with a constant velocity, v. A combined model incorporating the fusion-pore detection error model and that for autoinhibition was then used to predict second and later quantal latencies, by using the first quantal latencies to determine the estimates for the parameters in the combined model. When this analysis was done on the times for quantal secretion at sites on thirteen different motor-nerve terminals, the value of v was estimated as zero in each case, so that no autoinhibitory effect was observed.

Effects of acrylamide on cotransmission in perivascular sympathetic and sensory nerves

The effects of chronic administration of acrylamide on sympathetic and sensory nerves were examined in the mesenteric artery of rabbits. The noradrenaline (NA) content of the artery was significantly decreased and the total contractile response to electrical field stimulation (4–64 Hz) markedly reduced in the acrylamide group. This was not due to an impairment of the contractility of the smooth muscle or to alterations in the postjunctional receptors. At 16 Hz, only the purinergic component of sympathetic cotransmission was significantly reduced by acrylamide. At 64 Hz, both the purinergic and the adrenergic components were significantly decreased. Field stimulation of the artery pretreated with guanethidine and precontracted with NA produced a frequency-dependent relaxation which was prevented by capsaicin and thus mediated by perivascular sensory nerves. In contrast to its effects on sympathetic cotransmission, acrylamide resulted in a trend, although not significant, towards increased responses at each frequency studied (2–16 Hz). 2-Methylthio-ATP (2Mc-S-ATP) caused significantly greater relaxation following acrylamide treatment while vasodilator responses to calcitonin gene-related peptide and substance P were unchanged. It is concluded that, in addition to its known action in producing neuropathy in myelinated somatic motor and sensory nerves, acrylamide causes damage to unmyelinated perivascular sympathetic fibres. Purinergic mechanisms may be particularly susceptible to acrylamide since both the purinergic component of sympathetic vasoconstriction and the relaxation in response to 2Mc-S-ATP were affected by acrylamide treatment.