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A pharmacological study of the effect of carbamazepine on neuromuscular transmission in the rat diaphragm.

The effects of carbamazepine (0.042-0.42 mM) on neuromuscular transmission were studied on the isolated rat phrenic nerve diaphragm preparation using standard pharmacological and electrophysiological methods. Carbamazepine decreased (1) the antidromic activity of the phrenic nerve, (2) the amplitude of the endplate potential (EPP) and miniature endplate potential (MEPP), (3) the quantal content of the endplate potential, (4) the indirectly-elicited twitch tension, (5) the muscle contracture in chronically denervated muscle induced by acetylcholine (ACh) and (6) the amplitude of the compound phrenic nerve action potential, in a concentration-dependent manner. The antidromic activity of the phrenic nerve was the most affected, while the phrenic nerve compound action potential was least affected. However, the IC50 for carbamazepine (the concentration of carbamazepine that inhibited 50% of the response) was in the same order of concentration, i.e. 0.11-0.3 mM. Compared with the effect of carbamazepine on the indirectly-elicited twitch tension with its actions described above, it is concluded that carbamazepine interfered with the neuromuscular activity by inhibiting pre- and postsynaptic process and conduction in the phrenic nerve.

Action Potentials↗

Dystrophin-related protein is localized to neuromuscular junctions of adult skeletal muscle.

Dystrophin-related protein (DRP) is an autosomal gene product with high homology to dystrophin. We have used highly specific antibodies to the unique C-terminal peptide sequences of DRP and dystrophin to examine the subcellular localization and biochemical properties of DRP in adult skeletal muscle. DRP is enriched in isolated sarcolemma from control and mdx mouse muscle, but is much less abundant than dystrophin. Immunofluorescence microscopy localized DRP almost exclusively to the neuromuscular junction region in rabbit and mouse skeletal muscle, as well as mdx mouse muscle and denervated mouse muscle. DRP is also present in normal size and abundance and localizes to the neuromuscular junction region in muscle from the dystrophic mouse model dy/dy. Thus, DRP is a junction-specific membrane cytoskeletal protein that may play an important role in the organization of the postsynaptic membrane of the neuromuscular junction.

Animals↗

Heme oxygenase-2 expression at rat neuromuscular junctions.

The neuromuscular junction is specialized for rapid transmission of electrical signals. Nitric oxide synthase (NOS) is concentrated at the junction, and NO modulates transmission and could influence signaling pathways. Increasing evidence suggests that carbon monoxide (CO) serves as a neurotransmitter, and heme oxygenase (HO), the enzyme that catalyzes the formation of CO, is often colocalized with NOS. Immunoreactivity for HO-2 was present at rat neuromuscular junctions of leg muscles and persisted in denervated muscle indicating the localization of the enzyme to the postsynaptic surface. In contrast, HO-2 immunoreactivity was absent from the en grappe and orbital en plaque endplates of extraocular muscle (EOM), while only the global en plaque endplates possessed HO-2 immunoreactivity. The difference between EOM and leg endplates may arise from EOM's unique physiology. The presence of HO-2 at neuromuscular junctions suggests CO could serve as a pre- and post-synaptic messenger.

Animals↗

Expression of an alpha-cardiac like myosin heavy chain in diaphragm, chronically stimulated, and denervated fast-twitch muscles of rabbit.

An additional slow fibre type, type I alpha, is detected in diaphragm and appears in fast-twitch hindlimb muscles of rabbit under the influence of altered neuromuscular activity. Type I alpha fibres were delineated from fibres expressing myosin heavy chain I beta (type I beta) by immunohistochemistry with a monoclonal antibody raised against the alpha-cardiac MHCI alpha. When stained for mATPase after acid and alkaline preincubations, some type I alpha fibres resembled type I beta and type IIA fibres, respectively. Some type I alpha fibres displayed dissimilar mATPase staining, indicating heterogeneity of this fibre population. The appearance of numerous type I alpha fibres in stimulated muscles, which in addition contain type IIA and type I beta fibres, suggested that they may be interspaced between types IIA and I beta. Electrophoresis under nondenaturing conditions disclosed an additional isomyosin both in normal diaphragm and stimulated muscles. This band displayed the same mobility as the slowest isomyosin in rabbit masseter muscle. It was recognized by the same monoclonal (anti-alpha-cardiac MHC) antibody used for immunohistochemistry. Therefore, this isomyosin appeared to be very similar, but perhaps not identical to the alpha-cardiac MHC-based isomyosin, probably resulting from discrete differences in the MHC complement. This assumption agrees with additional findings suggesting an even greater heterogeneity of the MHCs than generally assumed. In support of this, we show in atrium and masseter muscles the existence of an additional, electrophoretically distinct MHC isoform which migrates in close vicinity to MHCI alpha.

Animals↗

A physiological correlate of disuse-induced sprouting at the neuromuscular junction.

Recent investigations have established that many of the normal properties of muscle fibres are maintained, at least in part, by muscle activity. Thus, a fall in resting membrane potential, an increase in input resistance, and spread of acetylcholine receptors to extrajunctional sites can all be induced by abolishing muscle activity and prevented by direct stimulation of denervated muscle fibres. Muscle activity also exerts a trophic influence on the innervating motoneurones; furthermore it may be a factor in the regulation of sprouting. Brown and Ironton found fine, "ultra-terminal sprouts" emanating from the endplates of muscles rendered inactive by chronic conduction block of the muscle nerve. Pestronk and Drachman saw increased branching of the motor nerve terminal and a consequent increase in endplate size in similar conditions. If these sprouts at the endplates of inactive muscles were functional, one might expect more transmitter to be released in response to nerve stimulation. We report here that both quantum content and spontaneous miniature endplate potential (m.e.p.p) frequency are increased at the terminals of inactive (disused) muscles.

Acetylcholine↗

Soluble phosphatidylinositol phosphodiesterase in normal and denervated fast and slow muscles of the rat.

Phosphatidylinositol phosphodiesterase activity was determined in cytosol prepared from rat slow (soleus) and fast (extensor digitorum longus) muscles. The substrate was prepared by incubation of sarcoplasmic reticulum with myo-[2-3H]inositol. The enzyme hydrolysed both membrane-bound and extracted phosphatidylinositol. The activity determined with the isolated phospholipid exhibited an optimum at pH 5.5. Ca2+ ions stimulated the activity. The enzyme specific activity was higher in cytosol prepared from soleus muscle than in that from extensor digitorum longus muscle. After section of the motor nerve, the activity of the enzyme increased in both muscles up to 36 h and then declined. A function for this enzyme in the control of acetylcholine sensitivity in muscle is discussed.

Animals↗

Phencyclidine interactions with the ionic channel of the acetylcholine receptor and electrogenic membrane.

The effects of phencyclidine (PCP) were studied on the electrogenic and chemosensitive properties of the neuromuscular junction of skeletal muscle as well as on the binding sites on the acetylcholine (AcCho) receptor and its ionic channel in the electric organ membranes of the electric ray. The directly elicited muscle twitch was markedly potentiated by prolonging the falling phase of the muscle action potential and blocking delayed rectification. The indirectly elicited muscle twitch was transiently potentiated and then blocked by PCP at concentrations below 60 muM. PCP blocked miniature endplate potentials and AcCho sensitivities at the junctional region of innervated muscle, blocked the extrajunctional sensitivity of the chronically denervated muscle, and significantly depressed the peak amplitude of the endplate current (EPC) in a voltage- and time-dependent manner. PCP also caused acceleration of the time course of EPC decay and shortening of the mean life-time of the open ionic channel. The effects of PCP were not due to inhibition of AcCho receptor sites because PCP did not protect against the quasi-irreversible inhibition of receptor sites by alpha-bungarotoxin, nor did it inhibit binding of [(3)H]AcCho or [(125)I-labeled alpha-bungarotoxin to the receptor sites. On the other hand, PCP blocked the binding of [(3)H]perhydrohistrionicotoxin to the sites of the ionic channel of the AcCho receptor. The data suggest that PCP reacts with the electrogenic K(+) channel and the ionic channel associated with the AcCho receptor in the open as well as the closed conformation.

Action Potentials↗

Neuronal activity of the cat supraoptic nucleus is influenced by muscle small-diameter afferent (groups III and IV) receptors.

In anesthetized cats, responses of single neurosecretory neurons of the supraoptic nucleus to activation of muscle receptors were investigated. Electrical stimulation (1-3 pulses at 200 Hz) of group III and IV pure muscle afferents (gastrocnemius nerve) evoked excitation of greater than 50% of supraoptic nucleus neurons (n = 50), whereas stimulation of group Ia or Ib fibers was ineffective. Baroreceptor stimulation inhibited 95% of these supraoptic nucleus neurons that responded to activation of muscle afferents. Excitation of receptors in the gastrocnemius muscle by intra-arterial injection of chemicals (NaCl, KCl, and bradykinin) increased firing rates of most (84%, 74%, and 80%, respectively) neurosecretary neurons. The magnitude of the excitatory response was dose dependent--bradykinin being the most effective. The response disappeared after muscle denervation. When the gastrocnemius muscle alone was contracted phasically by ventral root stimulation, discharges of the supraoptic nucleus neurons increased, whereas quick stretch of the muscle had no effect. We conclude that activation of muscle receptors by chemical or mechanical stimulus can directly excite neurosecretory neurons in the supraoptic nucleus and that afferent impulses are carried by polymodal fibers of small diameter but not by the largest afferents (group I) from the muscle. The results may relate to increased concentrations of plasma vasopressin during exercise.

Animals↗

The effect of botulinum toxin type-A injection on spasticity, range of motion and gait patterns in children with spastic diplegic cerebral palsy: an Egyptian study.

Spasticity is defined as increased resistance to passive movement, secondary to hyperreflexia after an upper motor neuron lesion. In children with cerebral palsy (CP), it can interfere with mobility and self-care and can contribute to development of fixed myostatic contractures. This study investigated the efficacy of botulinum toxin type-A, a neuromuscular blocking agent that reduces muscle tone, in a variety of neuromuscular disorders, injections in a prospective, 3-month, controlled study involving 40 children with spastic diplegic CP. The patients were divided into two groups: Group 1 (20 patients) entered a botulinum toxin type-A injection+physiotherapy rehabilitation program; Group 2 (20 patients) were given the physiotherapy rehabilitation program only. Patients were assessed at 4, 8 and 12 weeks post-treatment using the Modified Ashworth Scale (MAS), dynamic gait pattern, ankle range-of-motion measurements and quantification of muscle denervation by nerve conduction techniques. The botulinum toxin type-A group demonstrated statistically significantly decreased spasticity, improved gait function and improved range of motion with evidence of partial denervation of the injected muscle compared to the control group. In conclusion, botulinum toxin type-A injections are a well-tolerated, non-surgical technique that can improve overall response to physiotherapy.

Ankle Joint↗