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.     

Acetylcholine noise: analysis after chemical modification of receptor

The elementary voltage pulses ("shot effects") produced by the action of acetylcholine molecules on the receptor were studied by analyzing the membrane voltage fluctuations ("noise") after acetylcholine application at the frog neuromuscular junction. The amplitude of these pulses was decreased after treatment with a disulfide-bond reducing agent. The shot effect may thus depend on the structure or conformation of the receptor molecule.     

Tetrodotoxin and manganese ion: effects on action potential of the frog heart

Tetrodotoxin, which effectively suppresses the permeability of various tissues to sodium ions, has no effect on the calcium action potential of the fiber of barnacle muscle, which potential is produced by increase in permeability to calcium ions. Manganese ions, however, suppress the action potential. When applied to fiber of frog cardiac ventricle, tetrodotoxin suppresses the rate of rise of the action potential without affecting the overshoot; the suppressive effect of manganese ion is mainly on the overshoot of the action potential. This suggests that, in the action potential of the car-diac ventricle of the frog, the platealu phase is related primarily to the increase in permeability of the membrane to calcium ions.     

伊犁马LPL基因多态性及其与肉质性状关联性分析

【目的】研究伊犁马脂蛋白脂酶(lipoprotein lipase,LPL)基因多态性及其对肉质性状的影响,为专门化肉用马选种提供技术支撑。【方法】以41匹成年伊犁马为研究对象,采集颈静脉血液样本及其三角肌、背阔肌、臂三头肌、腹外斜肌和臀中肌5个部位肌肉样本,提取伊犁马DNA样本并测定肉质性状,利用PCR测序法筛选LPL基因突变位点并进行多态性分析,结合伊犁马肉质性状数据与LPL基因多态性进行关联性分析。【结果】所选马匹中共检测出2个突变位点(g.715T>C和g.12074C>A),均为内含子突变,2个位点均只检测出2种基因型,为低度多态位点。g.715T>C位点TT基因型马匹背阔肌和臀中肌失水率显著高于TC基因型马匹(P<0.05),背阔肌剪切力极显著大于TC基因型马匹(P<0.01),腹外斜肌剪切力极显著小于TC基因型马匹(P<0.05),三角肌和臀中肌剪切力显著大于TC基因型马匹(P<0.05);g.12074C>A突变位点CC基因型马匹臂三头肌和臀中肌失水率极显著低于CA基因型马匹(P<0.01),背阔肌和腹外斜肌失水率显著低于CA基因型马匹(P<0.05),臂三头肌熟肉率极显著高于CA基因型马匹(P<0.01),三角肌和腹外斜肌熟肉率与CA基因型马匹呈显著差异(P<0.05),臂三头肌和腹外斜肌剪切力极显著高于CA基因型马匹(P<0.01)。【结论】LPL基因g.715T>C和g.12074C>A位点是影响伊犁马肉质性状的潜在位点。     

Acetylcholine sensitivity of muscle fiber membranes: mechanism of regulation by motoneurons

Inhibitors of RNA and protein synthesis prevent the development of acetylcholine supersensitivity in organ-cultured rat diaphragm muscle but do not affect established acetylcholine sensitivity. The restriction of this sensitivity in innervated muscle apparently involves muscle fibers.     

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。     

EFFECT OF SPINAL FLUID FROM PATIENTS WITH MYASTHENIA GRAVIS ON THE SYNTHESIS OF ACETYLCHOLINE IN VITRO

Human spinal fluid is a more favorable medium to further the synthesis of acetylcholine in vitro, using enzyme obtained from frog brain, than is serum. Also, since less acetylcholine was synthesized in the presence of spinal fluid from patients with myasthenia gravis than spinal fluid of control subjects, it is probable that at least some of the factors responsible for the decrease and increase of the synthesis of acetylcholine pass into the spinal fluid.     

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.     

Solute concentration gradients in frog muscles at 0 degree C: active transport or adsorption?

In isolated frog muscle that has been incubated for 7 days at 0 degrees C invitro K(+) and Na(+) remain at normal concentrations. Amino acids are accumulated against a concentration gradient at this temperature; for example, glycine accumulates in muscle to a concentration ten times that in the external solution. The amount of cycloleucine accumulated is greater at 0 degrees C than at 25 degrees C. These findings, which are difficult to explain on the basis of metabolically linked active transport, are Consiststent with the view that solute accumulation by cell is the result of adsorption on spesific sites.     

Turbidity, birefringence, and fluorescence changes in skeletal muscle coincident with the action potential

Electrical stimulation of frog striated muscle was found to prduce trasient changes in turbidity. birefringence, and fluorescence (of fibers stained with pyronine B). The initial phase of these optical changes Was coinicident with the action potential. These findings suggest that there is a macromolecular conformational change in muscle membrane during excitation.     

Acetylcholine-synthesizing T cells relay neural signals in a vagus nerve circuit

Neural circuits regulate cytokine production to prevent potentially damaging inflammation. A prototypical vagus nerve circuit, the inflammatory reflex, inhibits tumor necrosis factor-α production in spleen by a mechanism requiring acetylcholine signaling through the α7 nicotinic acetylcholine receptor expressed on cytokine-producing macrophages. Nerve fibers in spleen lack the enzymatic machinery necessary for acetylcholine production; therefore, how does this neural circuit terminate in cholinergic signaling? We identified an acetylcholine-producing, memory phenotype T cell population in mice that is integral to the inflammatory reflex. These acetylcholine-producing T cells are required for inhibition of cytokine production by vagus nerve stimulation. Thus, action potentials originating in the vagus nerve regulate T cells, which in turn produce the neurotransmitter, acetylcholine, required to control innate immune responses.     

Hemicholinium-3: noncholinergic effects on squid axons

Hemicholinium-3, when applied to the inside of a squid axon, is effective in blocking the action potential. This action is not antagonized by the addition of choline or acetylcholine to the perfusate. Voltage-clamp experiments show that hemicholinium-3 depresses both the early transient and late steady-state components of membrane ionic conductances, with a greater effect on the peak transient component.     

Neurotransmitter release from a vertebrate neuromuscular synapse affected by a food dye

The food dye erythrosine (Erythrosin B; FD & C No. 3) was applied to isolated neuromuscular synapses in the frog, and its effects on the spontaneous quantal release of acetylcholine were examined with electrophysiological techniques. At concentrations of 10 muM or greater this anionic dye produced an irreversible, dose-dependent increase in neurotransmitter release. This increase did not depend on the presence of calcium ions in the bathing medium. These increase did not depend on the presence of calcium ions in the bathing medium. These results suggest that erythrosine might prove a useful pharmacological tool for studying the process of transmitter release, but that its use as a food additive should be reexamined.     

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.     

Choline acetyltransferase activity is increased in combined cultures of spinal cord and muscle cells from mice

The activity of choline acetyltransferase was more than tenfold greater in combined cultures of spinal cord and muscle cells than in cultures of spinal cord cells alone. This increase was associated with the formation of functional neuromuscular junctions in culture. Counts of silver-stained cells and determinations of other enzyme activities indicated that the increased choline acetyltransferase activity was not due to nonspecific neuronal survival but reflected greater activity in the surviving neurons. Hence, muscle had a marked, highly specific trophic effect on the cholinergic neurons that innervated it.     

Generation of adrenergic and cholinergic potentials in sympathetic ganglion cells

Norepinephrine elicited a hyperpolarizing response, and acetylcholine (during nicotinic blockade) elicited a depolarizing one. Both responses showed no increase in membrane conductance. The norepinephrine response was suppressed by initial depolarization; the acetylcholine response (frog cells); by hyperpolarization. These neurotransmitters apparently can activate electrogenic mechanisms which do not involve movement of ions down their electrochemical gradients.     

Reinnervated eye muscles do not respond to impulses in foreign nerves

Normal movements return to carp eyes after section and regeneration of the IIIrd and IVth nerve trunks. Two months after reinnervation, records of impulses in the inferior oblique nerve during tilting of the body show activity of the normal motoneurons to that muscle, together with discharge patterns characteristic of the antagonistic superior oblique and some of the rectus muscles. These axons must have found their way into the inferior oblique trunk during sprouting at the lesion and must be maintained after reinnervation. Impulses from foreign axons are without detectable effect on eye movement and therefore must be blocked at their termination in the muscle. Previous study of cross-innervated and doubly innervated fish eye muscles revealed only structurally normal neuromuscular junctions. Transmission from foreign junctions in multiply innervated muscle is blocked by competitive molecular recognition and control mechanisms that do not cause degeneration.     

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.     

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.