title: Syntaxin 1A occludes GABA(B) receptor-induced inhibition of exocytosis downstream of Ca(2+) entry in mouse hippocampal neurons.
abstract: The mechanisms underlying gamma-amino butyric acid (GABA(B)) receptor-mediated inhibition of exocytosis have been characterized in a variety of synapses. Using patch-clamp recording methods, we attempted to clarify the intracellular mechanisms underlying presynaptic inhibition in autaptic synapses of isolated mouse hippocampal neurons. Baclofen, a selective GABA(B) receptor agonist, decreased the frequency of glutamatergic miniature excitatory postsynaptic currents (mEPSCs) without changing their amplitude in Ca(2+)-free extracellular solution, suggesting that baclofen inhibits exocytosis downstream of Ca(2+) entry. Syntaxin 1A is known to modulate exocytosis and suppress neuronal sprouting. Antisense oligonucleotide-mediated knockdown of syntaxin 1A increased the frequency of mEPSCs under Ca(2+)-free condition. Estimation of the number of functional release sites by staining with FM1-43 indicated that the increased frequency of mEPSCs was induced by facilitation of exocytosis at each site, rather than by an increased number of release sites due to neuronal sprouting. Baclofen reduced mEPSC frequency in syntaxin 1A-knockdown neurons to the same level as that in nonsense oligonucleotide transfected neurons under Ca(2+)-free condition. These results suggest that the GABA(B) receptor- and syntaxin 1A-induced inhibitions of exocytosis occlude one another and that the GABA(B) receptor shares a common intracellular pathway with syntaxin 1A in inhibiting transmitter release downstream of Ca(2+) entry.
authors.affiliation: Department of Physiology and Biological Information, Dokkyo Medical University School of Medicine, 880 Kitakobayashi, Mibu, Tochigi 321-0293, Japan. tfukushi@dokkyomed.ac.jp
authors.forename: Teruyuki
authors.initials: T
authors.lastname: Fukushima
authors.forename: Masayuki
authors.initials: M
authors.lastname: Tsuda
authors.forename: Toshiki
authors.initials: T
authors.lastname: Otsubo
authors.forename: Yuuichi
authors.initials: Y
authors.lastname: Hori
mesh_terms: D000818:Animals; D001418:Baclofen / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism* / Q000494:pharmacology; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004622:Embryo, Mammalian; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology; D019706:Excitatory Postsynaptic Potentials / Q000187:drug effects / Q000502:physiology; D005089:Exocytosis / Q000187:drug effects / Q000502:physiology*; D018755:GABA Agonists / Q000494:pharmacology; D006624:Hippocampus / Q000166:cytology*; D066298:In Vitro Techniques; D051379:Mice; D009474:Neurons / Q000187:drug effects / Q000502:physiology*; D020320:Oligoribonucleotides, Antisense / Q000494:pharmacology; D018408:Patch-Clamp Techniques / Q000379:methods; D011726:Pyridinium Compounds / Q000378:metabolism; D000644:Quaternary Ammonium Compounds / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis; D018080:Receptors, GABA-B / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D050827:Syntaxin 1 / Q000235:genetics / Q000378:metabolism*
chemical_list: D018691:Excitatory Amino Acid Antagonists; C073804:FM1 43; D018755:GABA Agonists; D020320:Oligoribonucleotides, Antisense; D011726:Pyridinium Compounds; D000644:Quaternary Ammonium Compounds; D012333:RNA, Messenger; D018080:Receptors, GABA-B; D050827:Syntaxin 1; D001418:Baclofen; D002118:Calcium