title: Presynaptic calcium-channel currents in normal and csp mutant Drosophila peptidergic terminals.
abstract: The study of regulated vesicle exocytosis, which underlies neurotransmitter and neuropeptide release, has benefited from a convergence of several independent approaches. These include the use of genetically tractable organisms and model preparations that allow a direct characterization of presynaptic ionic currents. Aiming for a comprehensive analysis of release, we had already developed a Drosophila preparation in which electrophysiological recordings from peptidergic terminals are feasible. Here, we report on the characterization of the Ca2+-channel currents present in these terminals. With Ba2+ as the charge carrier, the presynaptic membrane expresses a current type with high-activation threshold and little inactivation. This current is blocked by verapamil and diltiazem at micromolar concentrations, it is relatively insensitive to nifedipine and completely resistant to non-L-type Ca2+-channel antagonists. As a comparison, we also analysed the pharmacology of high-threshold Ba+2 currents on muscle fibres. A high-activation threshold Ca2+-channel current is also present in muscle fibres, albeit with a distinct pharmacological profile. Thus, peptidergic terminals and muscle fibres exhibit different subtypes of voltage-gated Ca2+ channels. The putative role of cysteine string protein (CSP) as a neuronal Ca2+-channel modulator was tested by examining the peptidergic presynaptic current in csp null mutants. We show that CSP is expressed in peptidergic boutons and abolished in the mutant. Direct recordings, under conditions that inhibit calcium influx into glutamatergic terminals, show that Ca2+-currents in peptidergic csp terminals are entirely normal. This result indicates that CSP is not a generic Ca2+-channel modulator and it might perform different functions in fast versus slow forms of release.
authors.affiliation: Instituto Cajal (CSIC), Madrid, Spain.
authors.forename: M
authors.initials: M
authors.lastname: Morales
authors.forename: A
authors.initials: A
authors.lastname: Ferrús
authors.forename: M
authors.initials: M
authors.lastname: Martínez-Padrón
mesh_terms: D000818:Animals; D001464:Barium / Q000493:pharmacokinetics / Q000502:physiology; D002118:Calcium / Q000493:pharmacokinetics; D002121:Calcium Channel Blockers / Q000494:pharmacology; D015220:Calcium Channels / Q000502:physiology*; D004110:Diltiazem / Q000494:pharmacology; D004330:Drosophila / Q000235:genetics*; D005089:Exocytosis / Q000502:physiology; D050956:HSP40 Heat-Shock Proteins; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D009132:Muscles / Q000294:innervation; D009474:Neurons / Q000737:chemistry / Q000502:physiology; D009543:Nifedipine / Q000494:pharmacology; D018408:Patch-Clamp Techniques; D017729:Presynaptic Terminals / Q000737:chemistry* / Q000378:metabolism; D014700:Verapamil / Q000494:pharmacology
chemical_list: D002121:Calcium Channel Blockers; D015220:Calcium Channels; D050956:HSP40 Heat-Shock Proteins; D008565:Membrane Proteins; C088460:cysteine string protein; D001464:Barium; D014700:Verapamil; D004110:Diltiazem; D009543:Nifedipine; D002118:Calcium