The PLC/IP 3 R/PKC pathway is required for ethanol-enhanced GABA release.
Research on the actions of ethanol at the GABAergic synapse has traditionally focused on postsynaptic mechanisms, but recent data demonstrate that ethanol also increases both evoked and spontaneous GABA release in many brain regions. Using whole-cell voltage-clamp recordings, we previously showed that ethanol increases spontaneous GABA release at the rat interneuron-Purkinje cell synapse. This presynaptic ethanol effect is dependent on calcium release from internal stores, possibly through activation of inositol 1,4,5-trisphosphate receptors (IP(3)Rs). After confirming that ethanol targets vesicular GABA release, in the present study we used electron microscopic immunohistochemistry to demonstrate that IP(3)Rs are located in presynaptic terminals of cerebellar interneurons. Activation of IP(3)Rs requires binding of IP(3), generated through activation of phospholipase C (PLC). We find that the PLC antagonist edelfosine prevents ethanol from increasing spontaneous GABA release. Diacylglycerol generated by PLC and calcium released by activation of the IP(3)R activate protein kinase C (PKC). Ethanol-enhanced GABA release was blocked by two PKC antagonists, chelerythrine and calphostin C. When a membrane impermeable PKC antagonist, PKC (19-36), was delivered intracellularly to the postsynaptic neuron, ethanol continued to increase spontaneous GABA release. Overall, these results suggest that activation of the PLC/IP(3)R/PKC pathway is necessary for ethanol to increase spontaneous GABA release from presynaptic terminals onto Purkinje cells.
Kelm
M Katherine
MK
Department of Pharmacology, School of Medicine, University of North Carolina, Chapel Hill, NC 27599-7365, USA. katie_kelm@unc.edu
Weinberg
Richard J
RJ
Criswell
Hugh E
HE
Breese
George R
GR
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002492:Central Nervous System Depressants / Q000494:pharmacology*; D002531:Cerebellum / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D004075:Diglycerides / Q000378:metabolism; D000431:Ethanol / Q000494:pharmacology*; D066298:In Vitro Techniques; D053496:Inositol 1,4,5-Trisphosphate Receptors / Q000378:metabolism*; D007395:Interneurons / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D008297:Male; D017729:Presynaptic Terminals / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011689:Purkinje Cells / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction; D010738:Type C Phospholipases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D005680:gamma-Aminobutyric Acid / Q000378:metabolism*
D002492:Central Nervous System Depressants; D004075:Diglycerides; D053496:Inositol 1,4,5-Trisphosphate Receptors; D000431:Ethanol; D005680:gamma-Aminobutyric Acid; D011493:Protein Kinase C; D010738:Type C Phospholipases; D002118:Calcium