Synaptic segregation at the developing neuromuscular junction

Throughout the developing nervous system, competition between axons causes the permanent removal of some synaptic connections. In mouse neuromuscular junctions at birth, terminal branches of different axons are intermingled. However, during the several weeks after birth, these branches progressively segregated into nonoverlapping compartments before the complete withdrawal of all but one axon. Segregation was caused by selective branch atrophy, detachment, and withdrawal; the axon branches that were nearest to the competitor's branches were removed before the more distant branches were removed. This progression suggests that the signals that mediate the competitive removal of synapses must decrease in potency over short distances.     

2 types of neuromuscular junction in the pharyngeal retractor muscle of snails

The pharyngeal retractor muscle of Helix lucorum is innervated by two symmetrical nerves which contain axons of two types forming myoneural junctions with the muscle cells. Type I junctions correspond to thick axons. These axon terminals which contain a large number of spherical, clear vesicles (41 +/- 5 nm) and a smaller number of dense-cored vesicles (67 +/- 3 nm) make contacts mainly with noncontractile sarcoplasmic processes of muscle cells. Type II junctions correspond to thin axons containing many of granules. Their axon terminals contact with contractile parts of muscle cells and contain a heterogenous population of vesicles: small spherical clear vesicles (44 +/- 2 nm), dense-cored vesicles and numerous irregularly outlined granules with fine-granular content (135 +/- 5 nm). Space between muscle cell is usually wide (50 nm and more) with the exception of sarcoplasmic processes where the gap may be less than 10 nm.     

Analysis of synaptic transmission in the neuromuscular junction using a continuum finite element model

There is a steadily growing body of experimental data describing the diffusion of acetylcholine in the neuromuscular junction and the subsequent miniature endplate currents produced at the postsynaptic membrane. To gain further insights into the structural features governing synaptic transmission, we have performed calculations using a simplified finite element model of the neuromuscular junction. The diffusing acetylcholine molecules are modeled as a continuum, whose spatial and temporal distribution is governed by the force-free diffusion equation. The finite element method was adopted because of its flexibility in modeling irregular geometries and complex boundary conditions. The resulting simulations are shown to be in accord with experiment and other simulations.     

Differential susceptibility of diaphragm muscle fibers to neuromuscular transmission failure

The pattern of glycogen utilization was used to determine whether various muscle fiber types in the rat diaphragm are differentially susceptible to neuromuscular transmission failure. Muscle segments from the midcostal region were repetitively stimulated directly or via the phrenic nerve at 10 or 75 Hz. Muscle fiber types were classified histochemically as type I, IIa, or IIb. The amount of muscle fiber glycogen depletion with direct stimulation depended on stimulation rate (75 Hz > 10 Hz) and fiber type (IIb > IIa > I). However, with nerve stimulation, muscle fiber glycogen depletion did not display the same dependency on stimulation rate (10 Hz > 75 Hz), although with stimulation at 10 Hz, the same rank order of fiber depletion was observed (IIb > IIa > I). This rank order of depletion was reversed (I > IIa > IIb) during repetitive stimulation of the nerve at 75 Hz. By intermittently stimulating the muscle directly during continuous nerve stimulation, we determined that neuromuscular transmission failure contributed significantly to the force decline after 2 min of stimulation at 75 Hz but relatively little to the force decline after 2 min of stimulation at 10 Hz. A significantly greater fraction of the force decline could be attributed to neuromuscular transmission failure with repetitive bouts of stimulation at 10 Hz. We conclude that neuromuscular transmission failure causes a significant portion of the force decline after 8 min of stimulation at 10 and 75 Hz, that all diaphragm fiber types are susceptible to neuromuscular transmission failure, but that type IIb fibers are particularly susceptible at higher frequencies of stimulation.     

Effects of cadmium and lead on adrenergic neuromuscular transmission in the rabbit

The effects of inorganic lead (PbCl2) and cadmium (DdCl2) on the pressor response of rabbit saphenous arteries produced by sympathetic nerve stimulation were examined. A 1- to 3-cm length of artery was removed, placed in a bath containing mammalian Ringer solution, and perfused with the same solution at a constant rate sufficient to maintain a 40-60 mmHg perfusion pressure. Increases in perfusion pressure resulting from electrical stimulation -f periarterial nerve endings were reduced or completely blocked by the addition of 5-20 muM lead or cadmium to the bathing solution for a period of 15-30 min. Responses to norepinephrine or to direct electrical stimulation of the muscle remained relatively unaffected. During lead or cadmium blockade, the response to nerve stimulation could be restored by a fourfold increase in calcium concentration. It is concluded that lead and cadmium reduce the response to sympathetic nerve stimulation primarily through an effect on presynaptic nerve terminals.     

Pathophysiology on the disorder of neuromuscular transmission defects

Neuromuscular transmission defects are often puzzling but challenging diseases for the pediatric and adult neurologists. Some of these disorders entail severe or even life-threatening disability: Most are treatable, but effective therapy requires precise diagnosis sometimes difficult to make especially in congenital defects. The diagnosis rests on the combination of clinical data, the electromyogram and additional studies that may include microelectrode analysis of neuromuscular transmission, ultrastructural and cytochemical studies of neuromuscular junction (NMJ) and biochemical/molecular genetic studies on muscle specimens. Understandably, these studies often depend on the collaboration of several investigators. In each myasthenic disorder, an abnormality affects neuromuscular transmission directly or causes secondary derangements that eventually affects transmission. Namely, lesions of both the presynaptic and/or postsynaptic area manifest a variety of clinical findings. Here was discussed the pathophysiology on myasthenia gravis and Lambert-Eaton myasthenic syndrome of abnormal autoimmunity, congenital myasthenic syndromes of a chromosomal defect, and poisoning of organophosphorus, botulism and some other toxins.     

Effects of mercuric chloride and methyl mercury on cholinergic neuromuscular transmission in the guinea-pig ileum

The effects of mercuric chloride (HgCl2) and methyl mercury (MeHg) were examined on basal mechanical activity and electrically-induced neurogenic cholinergic contractions (twitch contractions) in longitudinal muscle-myenteric plexus strips from guinea-pig distal ileum. Both compounds at 0.33 microM slightly enhanced the amplitude of twitch contractions in approximately 50% preparations. This effect was probably due to facilitation of acetylcholine (ACh) release since 0.1 and 1 microM mercurials increased electrically-evoked tritium outflow from [3H]choline preloaded muscle layer with attached myenteric plexus. Conversely, higher mercury concentrations inhibited twitch contractions (HgCl2 IC50 = 21.3 +/- 6.4 microM; MeHg IC50 = 45.1 +/- 5.5 microM), as well as contractions to exogenous ACh (0.1 microM) in resting preparations, and concomitantly increased the basal tone. The former effects possibly reflected an antimuscarinic activity of mercury, while the latter was related to alterations of calcium homeostasis in the effector cells. Indeed, the effect of HgCl2 on basal tone was antagonized by the Ca2+ entry blocker nifedipine (3, 10, 30 nM), indicating Hg-induced facilitation of Ca2+ influx through voltage-dependent channels. On the whole, our results suggest that cholinergic neuromuscular transmission and Ca(2+)-dependent mechanisms underlying smooth muscle contractility are targets for mercury toxicity in the intestine.     

Neural activity affects distribution of glutamate receptors during neuromuscular junction formation in Drosophila embryos

The prevalence of diseases associated with obesity, such as cardiovascular disease and diabetes mellitus, is higher in the spinal cord injury (SCI) population. Specifically, the mortality rate for cardiovascular disease is 228% higher in the SCI population. In addition, 100% of SCI individuals have osteoporosis in the paralysed extremities. These diseases are related to physical activity level, the level of the spinal cord lesion, and time post injury. Physically active SCI men and women have above-average fat mass (16 to 24% and 24 to 32%, respectively, compared with 15% for able-bodied men and 23% for able-bodied women), while sedentary SCI individuals have 'at-risk' levels of body fat (above 25% and 32%, respectively). The proportions and densities of the 3 main constituents comprising the fat-free body (mineral, protein and water) are altered following SCI. Bone mineral content decreases by 25 to 50%, and the magnitude of reduction is dependent on the level, completeness and duration of SCI. Because of denervation resulting in skeletal muscle atrophy, total body protein reduces by 30%, and total body water relative to bodyweight decreases by 15% following SCI. Indirect methods based on 2-component body composition models assume constant proportions and densities of mineral, protein, and water in the fat-free body. As a result, prediction equations based on 2-component models yield invalid estimates of fat and fat-free mass in the SCI population. Therefore, future research needs to directly quantify the proportions and densities of the constituents of the fat-free body in the SCI population relative to age, sex, physical activity level, level of the spinal cord lesion and time post injury, and to develop equations based on multicomponent body composition models.     

Contribution of mechanical activity and electrical activity to cellular electrical uncoupling in ischemic rabbit papillary muscle

We separated mechanical and electrical activity during ischemia and we assessed the contribution of each of these variables to ischemia induced cellular electrical uncoupling. In two groups of rabbit papillary muscles, mechanical and electrical activity at the onset of ischemia was abolished. We determined how this affected the time of onset of uncoupling. Also, action potential duration (APD80), conduction velocity and extracellular pH were recorded. In the first group, the BDM group, mechanical arrest was achieved with 10 mM BDM (2,3-butanedione monoxime). This had no effect on APD80 and conduction velocity. In the second group, the quiescent group, electrical as well as mechanical arrest was obtained by not stimulating the muscles. The effect of electrical arrest was estimated as the difference between this group and the BDM group. Mechanical arrest delayed uncoupling significantly (control: 14.6 +/- 0.9 (min), mean +/- S.E.M., n = 10; BDM: 19.3 +/- 1.1, n = 10; P < 0.025). Electrical arrest caused an even greater delay (quiescent: 41.6 +/- 4.6, n = 7; P < 0.001 v control and BDM). In the BDM group, these effects were not associated with a difference of APD80 during ischemia, but with a smaller decrease of conduction velocity. Delay of uncoupling correlated with delay of extracellular acidosis. We conclude: (1) mechanical and electrical activity both are significant determinants of ischemia induced uncoupling; (2) delay of uncoupling is associated with delay of extracellular acidosis, indicating a reduced rate of metabolism.     

Nerve activity but not intracellular calcium determines the time course of endocytosis at the frog neuromuscular junction

We used FM1-43 imaging and intracellular recordings of synaptic potentials to measure the time course of endocytosis in frog motor nerve terminals following tetanic nerve stimulation, and we used fura-2 imaging of intraterminal Ca2+ concentration to compare endocytic rate and [Ca2+]i. Following a 30 Hz tetanus, endocytosis declined exponentially with a time constant that depended on the duration of stimulation. The level of [Ca2+]i rose from a resting value of about 100 nM to more than 500 nM during 30 Hz stimulation, and rapidly declined to 200-250 nM after stimulation. [Ca2+]i returned to resting level with a time course that, like endocytosis, depended on the duration of tetanic stimulation. However, the rate of [Ca2+]i recovery was much slower than the rate of endocytosis, leading to the conclusion that endocytic rate is not determined solely by the instantaneous level of [Ca2+]i.     

An improved method for the simultaneous demonstration of mRNA and esterase activity at the human neuromuscular junction

Vulvar vestibulitis, a subset of vulvodynia, is present in 15% of patients in a general gynecologic practice. Only a few studies have focused on pathologic features of vulvar vestibulitis and none have included a control group. Punch biopsies from the vulvar vestibule of 12 patients with an age range of 22 to 51 years (mean 28 years) and 12 age-matched controls were analyzed for histopathologic features and investigated for the role of probable etiologic factors including human papillomavirus (HPV). A chronic inflammatory infiltrate was present in all specimens from patients with vestibulitis, and was composed predominantly of T-lymphocytes with a small number of B cells and an admixture of plasma cells, mast cells, and occasional monocytes. T-helper suppressor ratio was normal. The infiltrate was mild in 5 patients, moderate in 1, and severe in 6. Minor vestibular glands were observed in 8 (66%) patients and were associated with a periglandular inflammatory infiltrate. Squamous metaplasia was observed in 4 (44%) patients. Epithelial hyperplasia was present in 10 (83%) patients with mild dysplasia in 2 (16%). Immunohistochemistry for immunoglobulins IgG, IgA, and IgM showed the presence of IgG-positive plasma cells in 75% of patients, suggesting chronic irritation, but an autoimmune etiology cannot be excluded or confirmed. Biopsies of control cases did not show any inflammatory infiltrate. In situ hybridization for HPV 6, 11, 16, and 18 was negative in the patient group as well as in the control group. We conclude that histopathologic abnormalities in patients with vulvar vestibulitis are the result of a chronic inflammatory reaction of the mucosa of the vulvar vestibule, for which the cause remains unclear.     

Formation of the neuromuscular junction: molecules and mechanisms

The vertebrate skeletal neuromuscular junction is the site at which motor neurons communicate with their target muscle fibers. At this synapse, as at synapses throughout the nervous system, efficient and appropriate communication requires the formation and precise alignment of specializations for transmitter release in the axon terminal with those for transmitter detection in the postsynaptic cell. Classical developmental studies demonstrate that synapse formation at the neuromuscular junction is a mutually inductive event; neurons induce postsynaptic differentiation in muscle cells and myofibers induce presynaptic differentiation in motor axon terminals. More recent experiments indicate that Schwann cells, which cap axon terminals, also play an active role in the formation and maintenance of the neuromuscular junction. Here, we review recent advances in the identification of molecules mediating such inductive interactions and the mechanisms by which they produce their effects. Although our discussion concerns events at developing neuromuscular junctions, it seems likely that similar molecules and mechanisms may act at neuron-neuron synapses in the peripheral as well as the central nervous system.     

Mucin histochemistry of the developing gastroesophageal junction

Mucin histochemistry was performed on the squamocolumnar junction (Z-line) of the gastroesophageal region of 49 autopsied previable fetuses, stillbirths, infants, and young children, using alcian blue pH 2.5/periodic acid-Schiff (PAS) and alcian blue pH 1.0/PAS stains. Sialylated and neutral mucins were present in most and sulfated mucin in many cases. In only one fetus was a heterotopic focus of goblet cells found in the distal esophagus. This study confirms that the presence of acidic mucins in columnar epithelium, without goblet cells, at the Z-line and adjacent cardia is common in this age group. Undue reliance on mucin stains to identify metaplastic columnar epithelium, in the absence of goblet cells, may result in overdiagnosis of Barrett's esophagus in children.     

Analysis of muscle electrical activity during indirect rhythmic stimulation

A mathematical model of transformation of the electrical signals in the peripheral neuromuscular junction, and a method of analysing the experimental data obtained in experiments on the muscle and muscle fibers of rat diaphragm in indirect rhythmic stimulation is described. The parameters characterizing the muscle by frequency potentiation and depression properties were determined as a result of the EMG analysis.     

Effects of nicotine on neocortical electrical activity in rats

The present study investigates the effects of acute and repeated nicotine i.p. treatment on cortical EEG activity. Nicotine at 0.3 and 0.9 mg/kg, but not at 0.1 mg/kg, decreased high voltage spindles (HVSs). Nicotine at 2.7 mg/kg suppressed HVSs completely. Mecamylamine, a nicotinic cholinergic antagonist, increased HVSs at 5 and 7.5 mg/kg. Nicotine blocked the HVS induction induced by mecamylamine. Mecamylamine at 1.25 mg/kg antagonized the HVS suppressing action of nicotine at 0.3 mg/kg. The muscarinic cholinergic antagonist, scopolamine (0.2 mg/kg), increased the 1 to 20 Hz amplitude sum value, and this increase was blocked to some extent by the highest dose of nicotine (2.7 mg/kg). However, nicotine did not block the effect of a higher scopolamine (2.0 mg/kg) dose on the sum amplitude values. Mecamylamine at 2.5 and 7.5 mg/kg blocked the effect of nicotine at 2.7 mg/kg on the EEG sum amplitude values in scopolamine (0.2 mg/kg)-treated rats. The peripherally acting nicotinic and muscarinic cholinergic antagonists, hexamethonium and scopolamine methylbromide, had no effect on spectral EEG and HVS values. In quisqualic acid nucleus basalis-lesioned rats, a frontal cortical choline acetyltransferase depletion (-72%) and slowing of the EEG was observed. Nicotine could not restore EEG activity in nucleus basalis-lesioned rats. After repeated (10 days, three injections/day) administration of nicotine, no tolerance to the effects of either nicotine (0.9 mg/kg) on spontaneously occurring HVSs or nicotine (2.7 mg/kg) on the EEG change induced by scopolamine was observed. The present results show that nicotinic receptor stimulation desynchronizes neocortical EEG activity in normal animals, but this action disappears in basal forebrain-lesioned animals. Therefore, it is likely that the effects of nicotine in reversing EEG and behavioral abnormalities observed in Alzheimer's disease may be limited if the basal forebrain cell loss is extensive.     

Effects of L-NG-nitro arginine on cholinergic transmission in the gastric muscle of the rabbit

In the circular muscle of the rabbit gastric corpus, the nitric oxide-synthesis inhibitor L-NG-nitro arginine (L-NOARG), enhanced the neurally-induced cholinergic responses evoked by electrical field stimulation (EFS) and ganglionic stimulating agents (nicotine, dimethylphenyl piperazinium iodide). The muscular contractions caused by acetylcholine (Ach) and methacholine were not influenced by the nitric oxide-synthesis inhibitor. The nitric oxide-releasing compound sodium nitroprusside (SNP) did not affect the Ach-induced muscular responses. Our results suggest that L-NOARG enhances gastric cholinergic responses by removing an inhibitory influence exerted at the prejunctional level in the nerve-muscle pathway.     

Effects of pemoline on spontaneous and event-related electrical activity of the brain

The effect of pemoline on the electrical activity of the brain (electroencephalogram, EEG) was studied in relation to time since sleep and time of day in 6 healthy subjects carrying out periods of work lasting 18 h. Power of the spontaneous EEG increased with time since sleep and amplitude of the P3 event-related response decreased. The changes may be interpreted as the reduction in alertness with time awake. In contrast, pemoline decreased power of the spontaneous EEG and increased the amplitude the P3 response, effects that are consistent with improved alertness. The changes in brain activity were paralleled by effects on performance, in terms of percentage of correct responses and reaction time. Performance decreased with time awake, and was improved by pemoline compared with placebo. The drug counteracted the adverse effects of time since sleep, with the beneficial effect of the drug persisting over the 18-hour period of work. The findings emphasise that spontaneous and event-related activity of the EEG may be used both to complement measures of performance in the laboratory and to assess behaviour in occupational situations where performance testing is impractical.     

Effects of neuromuscular blocking agents on excitatory transmission and gamma-aminobutyric acidA-mediated inhibition in the rat hippocampal slice

BACKGROUND: Although neuromuscular blocking agents do not cross the blood-brain barrier, they may penetrate the central nervous system under particular circumstances and eventually cause neurotoxic consequences. METHODS: The effects of neuromuscular blocking agents on excitatory and inhibitory transmission in area CA1 of rat hippocampal slices were investigated using extracellular and intracellular recording techniques. RESULTS: Application of atracurium in the perfusion medium resulted in a dose-dependent enhancement of excitatory synaptic responses averaging 48.7 +/- 4.3% at a concentration of 10 nM. This effect was correlated with an increase in the size of the presynaptic fiber volley. Laudanosine, but not pancuronium bromide or vecuronium bromide, produced similar changes. In addition, atracurium and laudanosine blocked inhibitory transmission and reduced intracellularly recorded gamma-aminobutyric acidA receptor-mediated potentials. These effects were observed only at concentrations >1 microM and were not reproduced by pancuronium bromide and vecuronium bromide. CONCLUSIONS: Atracurium and its metabolite, laudanosine, contrary to pancuronium bromide and vecuronium bromide, produce two distinct effects on hippocampal slices. They enhance excitatory transmission and neuronal excitability and they block inhibitory gamma-aminobutyric acidA-mediated synaptic responses.     

Analysis of anticholinesterase-induced neuromuscular transmission failure

To define the underlying mechanism of neuromuscular transmission failure induced by anticholinesterases, we simultaneously performed surface recordings of compound muscle action potentials (CMAPs) and intracellular recordings of miniature end-plate potentials (MEPPs), miniature end-plate current (MEPCs), and end-plate potential (EPPs) in rat diaphragms exposed in vitro to 1 x 10(-4) to 2 x 10(-2) mmol/L neostigmine methylsulfate. At low concentrations of neostigmine, repetitive stimulation of the phrenic nerve resulted in decrement followed by complete recuperation of CMAP amplitudes. This bimodal pattern was associated with maximal end-plate depolarization at the beginning of the stimulation period, increased MEPP amplitudes, and prolonged time constants of MEPC decays. Higher concentrations of neostigmine resulted in a unimodal decline of amplitudes of CMAPs and EPPS, reduced MEPP amplitudes, and a double exponential time course of MEPC decays. These results indicate that low concentrations of anticholinesterases impaired neuromuscular transmission by producing transient depolarization of the end-plate region. Higher concentrations induced desensitization and direct blockade of the end-plate receptor channel, probably in its open conformation.