title: Blockade of acetylcholine release at the motor endplate by a polypeptide from the venom of Phoneutria nigriventer.
abstract: 1. The mechanisms underlying the muscle relaxation effect of a fraction (PF3) isolated from the Phoneutria nigriventer spider venom were assessed on mouse diaphragm and chick biventer cervicis muscle preparations. 2. PF3 (0.25-4 micrograms ml-1) produced a concentration-dependent blockade of the nerve-elicited muscle twitch of the mouse diaphragm (IC50 = 0.8 micrograms ml-1) without affecting the directly induced muscle twitch. In similar preparations, the crude venom (1-10 micrograms ml-1) produced muscle contracture and blocked both the direct and indirectly induced muscle twitches. 3. In the chick biventer cervicis muscle, PF3 (1-5 micrograms ml-1) blocked the nerve stimulated muscle twitch (IC50 = 1.26 micrograms ml-1), but did not alter the postjunctional response to exogenous acetylcholine (ACh, 10 microM-10 mM). 4. PF3 (2-8 micrograms ml-1) reduced the frequency of miniature endplate potentials (m.e.p.ps) recorded intracellularly from the mouse diaphragm muscle fibers by 58 to 64%, and diminished the amplitude of m.e.p.ps by 20 to 40% of control. The relationship between log m.e.p.p. frequency and log [Ca2+]o was shifted rightwards in the presence of 4 micrograms ml-1 PF3. 5. Raising the frequency of m.e.p.ps with high K+ medium or theophylline (3 mM) did not prevent the toxin-induced depression of spontaneous ACh release. 6. The quantal content of e.p.ps (m), determined in cut-diaphragm muscle fibres, was reduced by 53% and 77% of control by 1 and 4 micrograms ml-1 PF3, respectively. At 1 microgram ml-1 the toxin shifted the relationship between log m and log [Ca2+]o towards higher values without apparent change of the slope. 7. E.p.p. trains elicited at 10 to 50 Hz in the presence of PF3 (1 microgram ml-1) exhibited irregular amplitudes and facilitation related to the frequency of nerve stimulation. 8. It is concluded that PF3 blocks neuromuscular transmission by acting prejunctionally and reducing the nerve-evoked transmitter release. The effect was related to a diminished Ca2+ entry into the nerve terminal associated with inhibition of exocytosis.
authors.affiliation: Universidade Federal de São Paulo, Escola Paulista de Medicina, Department of Pharmacology, São Paulo, Brazil.
authors.forename: C
authors.initials: C
authors.lastname: Souccar
authors.forename: M do C
authors.initials: Mdo C
authors.lastname: Gonçalo
authors.forename: A J
authors.initials: AJ
authors.lastname: Lapa
authors.forename: L R
authors.initials: LR
authors.lastname: Troncone
authors.forename: I
authors.initials: I
authors.lastname: Lebrun
authors.forename: F
authors.initials: F
authors.lastname: Magnoli
mesh_terms: D000109:Acetylcholine / Q000378:metabolism*; D000818:Animals; D002645:Chickens; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D009045:Motor Endplate / Q000187:drug effects*; D009126:Muscle Relaxation / Q000187:drug effects*; D009479:Neuropeptides / Q000494:pharmacology*; D009498:Neurotoxins / Q000494:pharmacology*; D011188:Potassium / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D013111:Spider Venoms / Q000737:chemistry*; D009435:Synaptic Transmission / Q000187:drug effects; D013806:Theophylline / Q000494:pharmacology
chemical_list: D009479:Neuropeptides; D009498:Neurotoxins; D013111:Spider Venoms; C098736:Tx3 neurotoxin; D013806:Theophylline; D000109:Acetylcholine; D011188:Potassium