GABA(A) receptors of cerebellar granule cells in culture: explanation of overall insensitivity to ethanol

• Published in: Neuroscience Vol. 162; no. 4; pp. 1187 - 1191
• Main Authors: Gatta, E, Cupello, A, Pellistri, F, Robello, M
• Format: Journal Article
• Language: English
• Published: United States 15.09.2009
• Subjects: Animals; Animals, Newborn; Cations, Divalent; Cells, Cultured; Cerebellum - cytology; Chloride Channels - physiology; Chlorides - pharmacology; Ethanol - pharmacology; Furosemide - pharmacology; GABA-A Receptor Antagonists; Neurons - drug effects; Neurons - physiology; Patch-Clamp Techniques; Protein Subunits - physiology; Rats; Receptors, GABA-A - physiology; Zinc Compounds - pharmacology
• ISSN: 1873-7544
• DOI: 10.1016/j.neuroscience.2009.05.031

GABA-activated chloride currents were studied in cerebellar granule cells put in culture from neonatal rats. As previously described, 10 microM GABA perfusion of these cells recorded by whole cell patch-clamp elicits chloride currents displaying a peak and a steady-state component. The two components were studied in the presence of 1 mM furosemide, 1 microM Zn(2+) and a combination of the two in order to evaluate the contribution of the different types of GABA(A) receptors. Furosemide inhibits alpha(6) containing receptors whereas low levels of Zn(2+) specifically block incomplete GABA(A) receptors made up of alpha and beta subunits only. The results show that the peak component involves the following receptors: alpha(x) beta(y), 25%; alpha(1) beta(y) gamma(2), 45%; alpha(6) beta(y) gamma(2) plus alpha(1) alpha(6) beta(y) gamma(2), 30%. The steady state component is made up by alpha(x) beta(y), 38%; alpha(1) beta(y) delta, 62%. Ethanol at relatively high concentration, 100 mM, slows further down the desensitization of alpha(1) beta(y) delta receptors. The results indicate that the relative insensitivity to ethanol of GABA(A) receptors of neonatal cerebellar granule cells in culture is due to the absence of mature alpha(6) beta(y) delta receptors, a major receptor brand involved in tonic inhibition.