Functional characterization of a kindling-like model of ethanol withdrawal in cortical cultured neurons after chronic intermittent ethanol exposure.
Chronic ethanol exposure has been reported to alter NMDA and GABA(A) receptor function and gene expression in brain regions of animals and mammalian cultured cortical neurons. In the present study, we investigated the effects of another model of chronic, but intermittent, ethanol treatment (CIE) on GABA(A) and NMDA receptor systems in cortical neurons. CIE (50 mM ethanol, 12 h exposure/12 h withdrawal, 5 cycles) exposure produced increased [3H]MK-801 binding and diazepam insensitive binding sites as measured by [3H]Ro15-4513 binding to cortical cultured neuronal membranes, at 0 h following the last treatment cycle relative to control neurons. The NMDA mediated increase in intracellular calcium [Ca2+]i was also increased following similar CIE treatment. CIE treatment also increased the ability of pentylenetetrazol (PTZ) to inhibit GABA mediated 36Cl- influx relative to control neurons. These effects were not reversible following 1 week ethanol withdrawal, implying enhanced sensitivity of PTZ to inhibit GABA(A) receptor mediated inhibition, and an increased NMDA receptor function in CIE treated cortical neurons. These alterations are consistent with the behavioral studies in animals, and suggest that both GABA(A) and NMDA receptors play an important role in ethanol withdrawal following either chronic or CIE exposure. Furthermore, this provides a feasible in vitro model for further biochemical and molecular studies of the mechanism underlying the CIE induced kindling-like phenomenon observed in humans.
Hu
X J
XJ
Department of Pharmacology, The University of Texas Health Science Center, San Antonio 78284, USA.
Ticku
M K
MK
D000818:Animals; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology / Q000187:drug effects*; D003975:Diazepam / Q000494:pharmacology; D004334:Drug Administration Schedule; D000431:Ethanol / Q000008:administration & dosage / Q000009:adverse effects*; D005260:Female; D018757:GABA Modulators / Q000494:pharmacology; D007696:Kindling, Neurologic; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008959:Models, Neurological; D009474:Neurons / Q000187:drug effects*; D010433:Pentylenetetrazole / Q000494:pharmacology; D013375:Substance Withdrawal Syndrome
D018757:GABA Modulators; D000431:Ethanol; D003975:Diazepam; D010433:Pentylenetetrazole