BDNF-dependent enhancement of exocytosis in cultured cortical neurons requires translation but not transcription.
Neurotrophic factors, such as brain-derived neurotrophic factor (BDNF), are involved in acute modulation of synaptic plasticity. Different modes of action of BDNF have been described with time courses ranging from seconds to hours, but the sequence of cellular processes responsible for BDNF-dependent modulation of synaptic plasticity is unknown. We have used optical imaging of the styryl dye, FM1-43, which selectively labels synaptic vesicles, to investigate potential presynaptic effects of BDNF. Addition of BDNF to cultured cortical neurons for 3 h produced a significant enhancement of exocytosis upon modest depolarization. BDNF had no effect on exocytosis either immediately or after incubation for 30 min. BDNF-dependent enhancement of exocytosis was blocked by the tyrosine kinase inhibitor, K252a, but not by K252b, consistent with signalling via the TrkB receptor. Having demonstrated that the BDNF-dependent enhancement of synaptic vesicle release was present only after 1 h, we investigated whether de novo gene transcription and/or protein synthesis were involved. Addition of the inhibitors of RNA synthesis, actinomycin D, or 5,6-dichloro-1-beta-D-ribofuranosyl benzimidazole (DRB), did not affect the enhancement of exocytosis produced by BDNF. However, the effect of BDNF was blocked by the inhibitors of translation, cycloheximide or anisomycin. Our results indicate a rapid BDNF-dependent enhancement of neurotransmitter release that requires translation but not transcription.
Bradley
J
J
Neurosciences Institute, San Diego, CA 92121, USA.
Sporns
O
O
D000818:Animals; D019208:Brain-Derived Neurotrophic Factor / Q000494:pharmacology*; D002478:Cells, Cultured; D004594:Electrophysiology; D005089:Exocytosis / Q000187:drug effects*; D005456:Fluorescent Dyes; D015870:Gene Expression / Q000502:physiology; D008564:Membrane Potentials / Q000187:drug effects; D009419:Nerve Tissue Proteins / Q000235:genetics; D009473:Neuronal Plasticity / Q000187:drug effects; D009474:Neurons / Q000166:cytology* / Q000187:drug effects / Q000502:physiology; D011188:Potassium / Q000494:pharmacology; D014176:Protein Biosynthesis / Q000502:physiology*; D011726:Pyridinium Compounds; D000644:Quaternary Ammonium Compounds; D051381:Rats; D011921:Rats, Inbred WKY; D009435:Synaptic Transmission / Q000187:drug effects; D013572:Synaptic Vesicles / Q000502:physiology; D013997:Time Factors; D014158:Transcription, Genetic