title: Increases in acetylcholine release produced by phorbol esters are not mediated by protein kinase C at motor nerve endings.
abstract: Recent work from our laboratory has demonstrated that phorbol esters known to stimulate protein kinase C (PKC) also stimulate acetylcholine (ACh) secretion by an action at a strategic component of the secretory apparatus [ J Physiol (Lond) 501:41-48]. In an attempt to determine whether the stimulatory effects of phorbols are mediated by PKC, we examined the effects of several PKC antagonists on ACh release promoted by phorbol 12,13-dibutyrate (PDBu) at the frog neuromuscular junction. PKC antagonists that act at the ATP binding site (C3 domain) were examined for their ability to antagonize the stimulatory action of PDBu. Neither the nonselective PKC inhibitor, staurosporine (at concentrations as high as 1 microM), nor its more selective derivative, GF109203X (at concentrations as high as 10 microM), attenuated the stimulatory effects of PDBu. PKC antagonists that act at the phorbol ester binding site (C1 domain) were examined for their ability to antagonize the stimulatory action of PDBu. Neither sphingosine (500 microM) nor calphostin C (25 microM) reduced the stimulatory actions of PDBu on ACh release. These results suggest that a presynaptic protein possessing a phorbol ester receptor and not the enzyme PKC is the target site for the stimulatory effects of phorbol esters at motor nerve endings.
authors.affiliation: Department of Molecular Pharmacology, Northwestern University Medical School, Chicago, Illinois 60611, USA.
authors.forename: T J
authors.initials: TJ
authors.lastname: Searl
authors.forename: E M
authors.initials: EM
authors.lastname: Silinsky
mesh_terms: D000109:Acetylcholine / Q000378:metabolism*; D000818:Animals; D002273:Carcinogens / Q000494:pharmacology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D009281:Naphthalenes / Q000494:pharmacology; D009469:Neuromuscular Junction / Q000187:drug effects*; D010703:Phorbol Esters / Q000494:pharmacology*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000187:drug effects* / Q000378:metabolism; D011894:Rana pipiens; D013110:Sphingosine / Q000494:pharmacology; D019311:Staurosporine / Q000494:pharmacology*
chemical_list: D002273:Carcinogens; D004791:Enzyme Inhibitors; D009281:Naphthalenes; D010703:Phorbol Esters; D011493:Protein Kinase C; D019311:Staurosporine; C058819:calphostin C; D000109:Acetylcholine; D013110:Sphingosine