Trophic regulation of acetylcholine sensitivity of muscle: effect of electrical stimulation

Denervation of skeletal muscle results in a spread of acetylcholine sensitivity over the entire surface membrane. Electrical stimulation, programmed to mimic the normal activity pattern, was applied continuously to the denervated rat diaphragm in vivo. After 4 days, the acetylcholine sensitivity was far less in the stimulated diaphragms than in denervated controls. Muscle activity may account for "neurotrophic" regulation of the acetylcholine sensitivity.     

Amantadine: neuromuscular blockade by suppression of ionic conductance of the acetylcholine receptor

Amantadine hydrochloride decreases the sensitivity of denervated mammalian muscle to iontophoretically applied acetylcholine. The drug depresses the amplitude of the end-plate current and reverses the slope of the relation between half-decay time and membrane potential suggesting that it alters the ionic conductance that is mediated by the acetylcholine receptor. Binding studies confirm that amantadine acts on the ion conductance modulator rather than the acetylcholine receptor.     

Acetylcholine: release from neural tissue and identification by pyrolysis--gas chromatography

Gas chromatographic analysis showed acetylcholine to be the only choline ester released from innervated longitudinal muscle of the guinea pigileum by electrical stimulation. The amount of acetylcholine measured by gas chromatography agreed almost exactly with that measured by bioassay. Denervated longitudinal muscle produced no acetylcholine, and treatment of the muscle with tetrodotoxin markedly reduced acetylcholine output. The method permits the recovery and quantitation of amounts of acetylcholine as low as 5 nanograms in 5 milliliters of tissue perfusates.     

Interaction with membrane remnants of target myotubes maintains transmitter sensitivity of cultured neurons

Parasympathetic neurons, when cultured alone, lose sensitivity to acetylcholine, but if striated muscle is included in the culture, neuronal chemosensitivity is maintained. The membrane remnants of myotubes ruptured by osmotic shock also supported the responsiveness of the cultured neurons to transmitter, whereas muscle-conditioned medium or membrane remnants of nonmuscle embryonic skin cells did not support this responsiveness. The regulation of chemosensitivity by contact of neurons with the target cell membrane may be important in the formation and maintenance of neuronal circuitry.     

Monoclonal antibody to acetylcholine receptor: cell line established from thymus of patient with Myasthenia gravis

A human B cell line producing a monoclonal antibody to an antigenic determinant of acetylcholine receptors was established by cloning B cells that had been transformed in vitro by Epstein-Barr virus. The B cells were obtained from the thymus of a patient with myasthenia gravis. The antibody produced by the cell line precipitated acetylcholine receptors from denervated and innervated rat muscle and from human muscle, but did not show detectable response to the acetylcholine receptors from the electric organs of Narke japonica. The monoclonal antibody showed identical binding patterns in innervated and denervated rat muscles. Passive transfer of the monoclonal antibody into rats induced moderate muscle weakness and electromyographic changes characteristic of myasthenia gravis.     

Modulation of acetylcholine receptor desensitization by forskolin is independent of cAMP

Biochemical and electrophysiological studies suggest that adenosine 3',5'-monophosphate (cAMP)-dependent phosphorylation of the nicotinic acetylcholine receptor channel is functionally significant because it modifies the receptor's rate of desensitization to acetylcholine. In studies that support this conclusion researchers have used forskolin to stimulate cAMP-dependent phosphorylation in intact muscle. It is now shown that although forskolin facilitated desensitization in voltage-clamped rat muscle, this effect was not correlated with the abilities of forskolin and forskolin analogs to activate adenylate cyclase or phosphorylate the receptor. Furthermore, elevation of intracellular cAMP or addition of the catalytic subunit of A-kinase failed to alter desensitization. Therefore, in intact skeletal muscle, cAMP-dependent phosphorylation does not modulate desensitization.     

ACETYLCHOLINE-LIKE ACTIVITY IN SCIATIC NERVE

Extracts of sciatic nerve exhibit acetylcholine-like activity that is only partly attributable to acetylcholine. The extracts show relatively greater activity on the rectus abdominis muscle of the frog than on the ileum of the guinea pig. To prevent the action of the extract on the frog rectus abdominis muscle, a greater concentration of d-tubocurarine is required than is necessary to prevent the action of known acetylcholine.     

影响獭兔离体肠道平滑肌收缩的因素

[目的]探讨影响獭兔离体肠道平滑肌收缩的因素。[方法]以獭兔十二指肠平滑肌为试验材料,以平滑肌收缩波的振幅和周期为评价指标,考察影响试验的各主要因素。[结果]在(36.5±0.5)℃台氏液中,獭兔离体肠管的存活时间和保质期最长,振幅最稳定,肾上腺素使离体肠管平滑肌收缩幅度减弱,乙酰胆碱、氢氧化钠及氯化钙均导致平滑肌收缩频率及幅度升高,收缩力增强。[结论]獭兔离体肠道平滑肌易受到温度、pH、其他化学因素及药物的影响。     

Insensitive Acetylcholine Receptor Conferring Resistance of Plutella xylostella to Nereistoxin Insecticides

The combinative rate measurement of (3-[Ⅰ125] iodotyrosyl) α-bungarotoxin was applied in the analysis of the relation between nerve acetylcholine receptor and three types of insecticide resistance in diamondback moth, Plutella xylostella (L.). In the dimehypo-resistant strain and in the cartap-resistant strain, the nerve acetylcholine receptor showed the remarkable insensitivity to dimehypo and cartap, of which the binding rate to ligand was approximately 66 and 60%, respectively, of the susceptible strain. The sensitivity to deltamethrin in the deltamethrin-resistant strain did not show visible change. These results indicated that the decline in the sensitivity of nerve acetylcholine receptor to insecticide might be a potential mechanism to nereistoxin insecticides resistance in the diamondback moth.     

小菜蛾nAChR靶标敏感性与沙蚕毒素类药物的抗性关系

【目的】研究抗杀虫双、杀螟丹和溴氰菊酯品系的神经乙酰胆碱受体及与它们对沙蚕毒素类药物的抗性间的关系。【方法】测定不同品系小菜蛾的神经乙酰胆碱受体与125I标记的α-银环蛇毒素的结合率。【结果】抗杀虫双、杀螟丹品系小菜蛾的神经乙酰胆碱受体对沙蚕毒素药物有显著的靶标不敏感性,它们与配体的结合率大约分别是敏感品系的66%和60%,在溴氰菊酯抗性品系中没有明显变化。【结论】神经乙酰胆碱受体对杀虫剂敏感性下降可能是小菜蛾对沙蚕毒素药物产生抗性的一种作用机理。     

Insensitive Acetylcholine Receptor Conferring Resistance of Plutella xylostella to Nereistoxin Insecticides

The combinative rate measurement of （3-[I125] iodotyrosyl） α-bungarotoxin was applied in the analysis of the relation between nerve acetylcholine receptor and three types of insecticide resistance in diamondback moth, Plutella xylostella （L.）. In the dimehypo-resistant strain and in the cartap-resistant strain, the nerve acetylcholine receptor showed the remarkable insensitivity to dimehypo and cartap, of which the binding rate to ligand was approximately 66 and 60%, respectively, of the susceptible strain. The sensitivity to deltamethrin in the deltamethrin-resistant strain did not show visible change. These results indicated that the decline in the sensitivity of nerve acetylcholine receptor to insecticide might be a potential mechanism to nereistoxin insecticides resistance in the diamondback moth.     

Transmitter sensitivity of neurons assayed by autoradiography

Ionic conductance channels that are opened by activating nicotinic acetylcholine receptors at synapses of sympathetic neurons are permeable to small organic amines. Uptake of a tritium-labeled amine through these channels can be measured by autoradiography. This provides a simple and direct way to assess the sensitivity of individual neurons to acetylcholine without using microelectrodes.     

Species specific effect of acetylcholine on bivalve rectums

The pharmacology of acetylcholine and 5-hydroxytryptamine on the rectums of Katelysia rhytiphora and K. scalarina was found to be similar, in general, to that of other bivalves. However, while acetylcholine causes a fast twitch in the rectum of K. rhytiphora, the activity and tone of the K. scalarina rectum are depressed except at high concentrations of the drug. The two species can be distinguished by these responses, and, therefore, these rectums are useful experimental objects for studying the physiology of molluscan visceral muscle.     

Cholinergic action of homogenates of sea urchin pedicellariae

A dialyzable, acetylcholine-like material exists in homogenates of globiferous pedicellariae of the sea urchin Lytechinus variegatus. Its pharmacological characterization was obtained on the basis of the responses of the guinea pig ileum, the rat uterus, blood pressure in the dog, the amphibian heart, the longitudinal muscle of the holothurian, and the protractor muscle of the sea urchin lantern. Some of the data were statistically treated, and an attempt was made to determine a ratio of potency between the homogenates and acetylcholine.     

Dok-7 mutations underlie a neuromuscular junction synaptopathy

Congenital myasthenic syndromes (CMSs) are a group of inherited disorders of neuromuscular transmission characterized by fatigable muscle weakness. One major subgroup of patients shows a characteristic "limb girdle" pattern of muscle weakness, in which the muscles have small, simplified neuromuscular junctions but normal acetylcholine receptor and acetylcholinesterase function. We showed that recessive inheritance of mutations in Dok-7, which result in a defective structure of the neuromuscular junction, is a cause of CMS with proximal muscle weakness.     

α6/α3β4乙酰胆碱受体在非洲爪蟾卵母细胞中的表达

α6/α3β4烟碱型乙酰胆碱受体（α6β4* nAChRs,*代表其他亚基）主要分布于大脑海马区、外周神经节和人源肾上腺嗜铬细胞等,与神经性疼痛和炎症反应、情绪等生理疾病密切相关。本研究拟对大鼠α6/α3β4 nAChR在非洲爪蟾（Xenopus laevis）卵母细胞膜上进行重组表达,并利用双电极电压钳电生理学技术检测该重组受体的功能。在不同浓度的激动剂乙酰胆碱的刺激下,比较了α6/α3和β4亚基的不同配比形成的受体通道开放而产生的电流大小,并用其拮抗剂α?芋螺毒素TxID验证了该受体的敏感性。结果表明,亚基不同配比的α6/α3β4 nAChR均可在非洲爪蟾卵母细胞膜上成功表达,并具有较好的配体门控敏感性。     

血根碱对兔离体肠平滑肌运动的舒张作用

【目的】探讨血根碱对兔离体肠平滑肌的舒张作用及机制。【方法】采用离体肠肌试验,应用BL-420E生物机能实验系统,比较血根碱对正常肠肌及工具药乙酰胆碱、酚妥拉明、氯化钡作用下兔离体肠平滑肌活动的影响,记录肠肌的收缩频率和振幅,并计算抑制率。【结果】血根碱可浓度依赖性的抑制兔小肠平滑肌自发性收缩的频率和振幅。0.4 mg•mL-1的血根碱对由乙酰胆碱、氯化钡引起的离体肠肌的兴奋性收缩有明显的拮抗作用,而对酚妥拉明阻断肾上腺素能受体后,仍能显著降低离体肠平滑肌收缩的振幅。【结论】血根碱对小肠平滑肌的舒张作用可能与乙酰胆碱竞争,阻断M受体、与酚妥拉明竞争,兴奋α受体及直接抑制肠肌兴奋性有关。     

Peptide neurotoxins from fish-hunting cone snails

To paralyze their more agile prey, the venomous fish-hunting cone snails (Conus) have developed a potent biochemical strategy. They produce several classes of toxic peptides (conotoxins) that attack a series of successive physiological targets in the neuromuscular system of the fish. The peptides include presynaptic omega-conotoxins that prevent the voltage-activated entry of calcium into the nerve terminal and release of acetylcholine, postsynaptic alpha-conotoxins that inhibit the acetylcholine receptor, and muscle sodium channel inhibitors, the mu-conotoxins, which directly abolish muscle action potentials. These distinct peptide toxins share several common features: they are relatively small (13 to 29 amino acids), are highly cross-linked by disulfide bonds, and strongly basic. The fact that they inhibit sequential steps in neuromuscular transmission suggests that their action is synergistic rather than additive. Five new omega-conotoxins that block presynaptic calcium channels are described. They vary in their activity against different vertebrate classes, and also in their actions against different synapses from the same animal. There are susceptible forms of the target molecule in peripheral synapses of fish and amphibians, but those of mice are resistant. However, the mammalian central nervous system is clearly affected, and these toxins are thus of potential significance for investigating the presynaptic calcium channels.     

Serum globulin in myasthenia gravis: inhibition of alpha-bungarotoxin binding to acetylcholine receptors

Serum factors that inhibit the binding of (125)I-labeled alpha-bungarotoxin to the acetylcholine receptor extracted in detergent from denervated rat muscle were detected by a sensitive assay. The serum of at least 5 and possibly 11 out of 15 patients with myasthenia gravis showed inhibitory activity that was localized to the globulin fraction. No controls showed inhibitory activity. The demonstration of inhibitory globulins may help explain the involvement of the immune system in the pathophysiology of the neuromuscular junction in patients with myasthenia gravis.     

低浓度甲氰菊酯对黄鳝乙酰胆碱酯酶活性的影响

[目的]了解甲氰菊酯对鱼类的毒副作用,为拟除虫菊酯类农药的污染监测提供参考。[方法]以低浓度甲氰菊酯（0.01、0.05、0.10、0.50、1.00μg/L）为供试药剂,采用接触染毒法研究不同作用时间下其对黄鳝肌肉乙酰胆碱酯酶（AChE）活性的影响。[结果]0.01μg/L甲氰菊酯对黄鳝肌肉AChE活性具有一定的诱导作用,随着甲氰菊酯浓度的升高,其对黄鳝肌肉AChE活性逐渐产生抑制作用,且其浓度越高对黄鳝肌肉AChE活性的抑制作用越强;连续暴露20d后,0.50、1.00μg/L甲氰菊酯处理组黄鳝肌肉AChE活性显著降低;随着染毒时间的延长,黄鳝肌肉AChE活性有逐渐降低的趋势。[结论]低浓度甲氰菊酯长时间处理可降低黄鳝肌肉AChE活性。