alpha-Bungarotoxin, kappa-bungarotoxin, alpha-cobratoxin and erabutoxin-b do not affect [3H]acetylcholine release from the rat isolated left hemidiaphragm.
Endplate preparations of the rat left hemidiaphragm were incubated with [3H]choline to label neuronal transmitter stores. Nerve evoked release of newly-synthesized [3H]acetylcholine was measured in the absence of cholinesterase inhibitors to investigate whether snake venom neurotoxins by blocking presynaptic nicotinic autoreceptors affect evoked transmitter release. Contractions of the indirectly stimulated hemidiaphragm were recorded to characterize the blocking effect of alpha-neurotoxins at the post-synaptic nicotinic receptors. Neither the long chain neurotoxins alpha-cobratoxin (1 microgram ml-1) and alpha-bungarotoxin (5 microgram ml-1) nor the short chain neurotoxin erabutoxin-b (0.1, 1 and 10 micrograms ml-1) affected the nerve-evoked release of [3H]acetylcholine. kappa-Bungarotoxin (1 and 5 micrograms ml-1), a toxin preferentially blocking neuronal nicotinic receptors, did also not affect evoked [3H]acetylcholine release, whereas (+)-tubocurarine (1 microM) under identical conditions reduced the release by about 50%. alpha-Bungarotoxin, alpha-cobratoxin and erabutoxin-b concentration-dependently (0.01-0.6 micrograms ml-1) inhibited nerve-evoked contractions of the hemidiaphragm. All neurotoxins except erabutoxin-b enhanced the basal tritium efflux immediately when applied to the endplate preparation or to a non-innervated muscle strip labelled with [3H]choline. This effect was attributed to an enhanced efflux of [3H]phosphorylcholine, whereas the efflux of [3H]choline and [3H]acetylcholine was not affected. It is concluded that the alpha-neurotoxins and kappa-bungarotoxin do not block presynaptic nicotinic receptors of motor nerves. These nicotinic autoreceptors differ from nicotinic receptors localized at the muscle membrane and at autonomic ganglia.
Apel
C
C
Department of Pharmacology, University of Mainz, Germany.
Rícný
J
J
Wagner
G
G
Wessler
I
I
D000109:Acetylcholine / Q000378:metabolism*; D000818:Animals; D002038:Bungarotoxins / Q000633:toxicity; D002851:Chromatography, High Pressure Liquid; D003039:Cobra Neurotoxin Proteins / Q000633:toxicity; D003964:Diaphragm / Q000187:drug effects* / Q000378:metabolism; D004870:Erabutoxins / Q000633:toxicity; D005260:Female; D007553:Isotope Labeling; D008297:Male; D009119:Muscle Contraction / Q000187:drug effects; D009130:Muscle, Smooth / Q000187:drug effects; D009498:Neurotoxins / Q000633:toxicity*; D010791:Phrenic Nerve / Q000187:drug effects*; D051381:Rats; D011978:Receptors, Nicotinic / Q000187:drug effects; D012910:Snake Venoms / Q000633:toxicity*; D009435:Synaptic Transmission / Q000187:drug effects*; D014316:Tritium / Q000378:metabolism
D002038:Bungarotoxins; D003039:Cobra Neurotoxin Proteins; D009498:Neurotoxins; D011978:Receptors, Nicotinic; D012910:Snake Venoms; D014316:Tritium; D004870:Erabutoxins; D000109:Acetylcholine