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J Neurophysiol 79: 1321-1328, 1998;
0022-3077/98 $5.00
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The Journal of Neurophysiology Vol. 79 No. 3 March 1998, pp. 1321-1328
Copyright ©1998 The American Physiological Society

Changes in Intracellular Calcium Concentration Affect Desensitization of GABAA Receptors in Acutely Dissociated P2-P6 Rat Hippocampal Neurons

Jerzy W. Mozrzymas and Enrico Cherubini

Biophysics Sector and Istituto Nazionale Fisica della Materia Unit, International School for Advanced Studies (SISSA), 34014 Trieste, Italy

Mozrzymas, Jerzy W. and Enrico Cherubini. Changes in intracellular calcium concentration affect desensitization of GABAA receptors in acutely dissociated P2-P6 rat hippocampal neurons. J. Neurophysiol. 79: 1321-1328, 1998. The whole cell configuration of the patch-clamp technique was used to study the effects of different cytosolic calcium concentrations [Ca2+]i on desensitization kinetics of gamma -aminobutyric acid (GABA)-activated receptors in acutely dissociated rat hippocampal neurons. Two different intrapipette concentrations of the calcium chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA; 11 and 0.9 mM, respectively) were used to yield a low (1.2 × 10-8 M) or a high (2.2 × 10-6 M) [Ca2+]i. In low [Ca2+]i, peak values of GABA-evoked currents (20 µM) evoked at -30 mV, were significantly larger than those recorded in high calcium [2,970 ± 280 (SE) pA vs. 1,870 ± 150 pA]. The extent of desensitization, assessed from steady-state to peak ratio was significantly higher in high calcium conditions (0.14 ± 0.007 vs. 0.11 ± 0.008). Similar effects of [Ca2+]i on desensitization were observed with GABA (100 µM). Recovery from desensitization, measured at 30 s interval with double pulse protocol was significantly slower in high [Ca2+]i than in low [Ca2+]i (54 ± 3% vs. 68 ± 2%). The current-voltage relationship of GABA-evoked currents was linear in the potential range between -50 and 50 mV. The kinetics of desensitization process including the rate of onset, extent of desensitization, and recovery were voltage independent. The run down of GABA-evoked currents was faster with the higher intracellular calcium concentration. The run down process was accompanied by changes in desensitization kinetics: in both high and low [Ca2+]i desensitization rate was progressively increasing with time as the slow component of the desensitization onset was converted into the fast one. In excised patches, the desensitization kinetics was much faster and more profound than in the whole cell configuration, indicating the involvement of intracellular factors in regulation of this process. In conclusion, [Ca2+]i affects the desensitization of GABAA receptors possibly by activating calcium-dependent enzymes that regulate their phosphorylation state. This may lead to modifications in cell excitability because of changes in GABA-mediated synaptic currents.




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