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Originally published In Press as doi:10.1074/jbc.M213081200 on February 24, 2003

J. Biol. Chem., Vol. 278, Issue 18, 16271-16279, May 2, 2003
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Declusterization of GABAA Receptors Affects the Kinetic Properties of GABAergic Currents in Cultured Hippocampal Neurons*

Enrica Maria PetriniDagger , Paola ZacchiDagger , Andrea BarberisDagger §, Jerzy W. Mozrzymas||, and Enrico CherubiniDagger **

From the Dagger  Neuroscience Programme and Istituto Nazionale Fisica della Materia Unit, International School for Advanced Studies, Via Beirut 2-4, Trieste 34014, Italy and the  Department of Biophysics, Wroclaw Medical University, ul. Chalubinskiego 10, Wroclaw 50-368, Poland

Speed and reliability of synaptic transmission are essential for information coding in neuronal networks and require the presence of clustered neurotransmitter receptors at the plasma membrane in precise apposition to presynaptic terminals. Receptor clusterization is the result of highly regulated processes involving functional and structural proteins. Among the structural elements, microtubules are known to play a crucial role in anchoring of gamma -aminobutyric acid, type A (GABAA) receptors. Here we show that microtubule depolymerization with nocodazole induces the declusterization of GABAA receptors and modifies the kinetic properties of GABAergic currents in cultured hippocampal neurons. In particular, this drug, applied either in the bath or via the patch pipette, induced the acceleration of the onset kinetics of miniature inhibitory postsynaptic currents (mIPSCs) without significantly affecting their frequency, thus suggesting a main postsynaptic site of action. After nocodazole treatment, current responses to ultrafast applications of GABA exhibited a faster rise time and an accelerated onset of desensitization. A quantitative analysis of GABA-evoked currents and model simulations suggest that declusterization affects the gating properties of GABAA receptors. In particular, a faster entry into the desensitized state of declustered GABAA receptors may account for the changes in the kinetic properties of mIPSCs after nocodazole treatment. Hence it appears that the clustered condition of GABAA receptors contributes in shaping GABAergic currents.

* This work was supported in part by the Ministero dell'Istruzione, dell'Universita' e della Ricerca (Grant COFI 2001 to E. C.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§ Present address: Dept. of Pharmacology and Toxicology, University of Zürich, Winterturerstrasse 190, Zürich CH-8057, Switzerland.

|| Supported by the Polish Committee for Scientific Research (Grant 6P04A-001-19).

** To whom correspondence should be addressed. Tel.: 39-040-378-7223; Fax: 39-040-378-7528; E-mail: cher@sissa.it.

Copyright © 2003 by The American Society for Biochemistry and Molecular Biology, Inc.
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