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J Neurophysiol 82: 3286-3297, 1999;
0022-3077/99 $5.00
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The Journal of Neurophysiology Vol. 82 No. 6 December 1999, pp. 3286-3297
Copyright ©1999 by the American Physiological Society

GABAA Receptor-Mediated Miniature Postsynaptic Currents and alpha -Subunit Expression in Developing Cortical Neurons

D. D. Dunning,1 C. L. Hoover,1 I. Soltesz,1 M. A. Smith,1 and D. K. O'Dowd1,2

 1Department of Anatomy and Neurobiology and  2Developmental and Cell Biology, University of California, Irvine, California 92697-1280

Dunning, D. D., C. L. Hoover, I. Soltesz, M. A. Smith, and D. K. O'Dowd. GABAA Receptor-Mediated Miniature Postsynaptic Currents and alpha -Subunit Expression in Developing Cortical Neurons. J. Neurophysiol. 82: 3286-3297, 1999. Previous studies have described maturational changes in GABAergic inhibitory synaptic transmission in the rodent somatosensory cortex during the early postnatal period. To determine whether alterations in the functional properties of synaptically localized GABAA receptors (GABAARs) contribute to development of inhibitory transmission, we used the whole cell recording technique to examine GABAergic miniature postsynaptic currents (mPSCs) in developing cortical neurons. Neurons harvested from somatosensory cortices of newborn mice showed a progressive, eightfold increase in GABAergic mPSC frequency during the first 4 wk of development in dissociated cell culture. A twofold decrease in the decay time of the GABAergic mPSCs, between 1 and 4 wk, demonstrates a functional change in the properties of GABAARs mediating synaptic transmission in cortical neurons during development in culture. A similar maturational profile observed in GABAergic mPSC frequency and decay time in cortical neurons developing in vivo (assessed in slices), suggests that these changes in synaptically localized GABAARs contribute to development of inhibition in the rodent neocortex. Pharmacological and reverse transcription-polymerase chain reaction (RT-PCR) studies were conducted to determine whether changes in subunit expression might contribute to the observed developmental alterations in synaptic GABAARs. Zolpidem (300 nM), a subunit-selective benzodiazepine agonist with high affinity for alpha 1-subunits, caused a reversible slowing of the mPSC decay kinetics in cultured cortical neurons. Development was characterized by an increase in the potency of zolpidem in modulating the mPSC decay, suggesting a maturational increase in percentage of functionally active GABAARs containing alpha 1 subunits. The relative expression of alpha 1 versus alpha 5 GABAAR subunit mRNA in cortical tissue, both in vivo and in vitro, also increased during this same period. Furthermore, single-cell RT-multiplex PCR analysis revealed more rapidly decaying mPSCs in individual neurons in which alpha 1 versus alpha 5 mRNA was amplified. Together these data suggest that changes in alpha -subunit composition of GABAARs contribute to the maturation of GABAergic mPSCs mediating inhibition in developing cortical neurons.




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