The amounts of mRNAs encoding α1, α6, β2, β3, γ2, and δ subunits of γ-aminobutyrate type A (GABA(A)) receptors and the gold immunolabeling density of their translation products were monitored during the growth of neonatal rat granule cells in primary culture. We investigated possible correlations (i) between temporal changes in mRNA content and expression density of their respective translation products and (ii) between the quantitative changes of receptor subunit expression, the GABA EC50 for Cl- channel activation, and diazepam efficacy in modulating GABA action on the Cl- channels. At 3 days in vitro, the amount of GABA(A) receptor subunit mRNAs and the expression of their respective translation products were very low. During the next 2 weeks both parameters for every subunit studied increased asynchronously; moreover, at 14 days in vitro the sum of γ2 and δ subunit expression was smaller than the expression of the α1 or α6 or β2/β3 subunits. This suggests that during in vitro maturation each subunit may be regulated independently and invites speculation as to possible changes in specific GABA(A) receptor subtype abundance during development in vitro. The maximal current intensity elicited by GABA failed to increase from 5 to 14 days in vitro, though the amount of mRNA encoding various subunits and the expression density of their respective translation products increased. Thus, qualitative changes in the GABA(A) receptor subtypes expressed and/or abnormalities in the subunit assembly very likely account for the uniformity of the maximal current intensity elicited by GABA during in vitro development. Also, during maturation of neuronal cultures from 5 to 20 days in vitro the extent of the positive modulation of GABA action by diazepam decreased dramatically. This finding might be related to an increase in the abundance of GABA(A) receptors including the α6 subunit and/or to the expression, during granule cell maturation in vitro, of GABA(A) receptors devoid of γ2 subunits.

Changes in γ-aminobutyrate type A receptor subunit mRNAs, translation product expression, and receptor function during neuronal maturation in vitro / Zheng, T. M.; Zhu, W. J.; Puia, G.; Vicini, S.; Grayson, D. R.; Costa, E.; Caruncho, H. J.. - In: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA. - ISSN 0027-8424. - 91:23(1994), pp. 10952-10956. [10.1073/pnas.91.23.10952]

Changes in γ-aminobutyrate type A receptor subunit mRNAs, translation product expression, and receptor function during neuronal maturation in vitro

Puia G.;
1994

Abstract

The amounts of mRNAs encoding α1, α6, β2, β3, γ2, and δ subunits of γ-aminobutyrate type A (GABA(A)) receptors and the gold immunolabeling density of their translation products were monitored during the growth of neonatal rat granule cells in primary culture. We investigated possible correlations (i) between temporal changes in mRNA content and expression density of their respective translation products and (ii) between the quantitative changes of receptor subunit expression, the GABA EC50 for Cl- channel activation, and diazepam efficacy in modulating GABA action on the Cl- channels. At 3 days in vitro, the amount of GABA(A) receptor subunit mRNAs and the expression of their respective translation products were very low. During the next 2 weeks both parameters for every subunit studied increased asynchronously; moreover, at 14 days in vitro the sum of γ2 and δ subunit expression was smaller than the expression of the α1 or α6 or β2/β3 subunits. This suggests that during in vitro maturation each subunit may be regulated independently and invites speculation as to possible changes in specific GABA(A) receptor subtype abundance during development in vitro. The maximal current intensity elicited by GABA failed to increase from 5 to 14 days in vitro, though the amount of mRNA encoding various subunits and the expression density of their respective translation products increased. Thus, qualitative changes in the GABA(A) receptor subtypes expressed and/or abnormalities in the subunit assembly very likely account for the uniformity of the maximal current intensity elicited by GABA during in vitro development. Also, during maturation of neuronal cultures from 5 to 20 days in vitro the extent of the positive modulation of GABA action by diazepam decreased dramatically. This finding might be related to an increase in the abundance of GABA(A) receptors including the α6 subunit and/or to the expression, during granule cell maturation in vitro, of GABA(A) receptors devoid of γ2 subunits.
1994
91
23
10952
10956
Changes in γ-aminobutyrate type A receptor subunit mRNAs, translation product expression, and receptor function during neuronal maturation in vitro / Zheng, T. M.; Zhu, W. J.; Puia, G.; Vicini, S.; Grayson, D. R.; Costa, E.; Caruncho, H. J.. - In: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA. - ISSN 0027-8424. - 91:23(1994), pp. 10952-10956. [10.1073/pnas.91.23.10952]
Zheng, T. M.; Zhu, W. J.; Puia, G.; Vicini, S.; Grayson, D. R.; Costa, E.; Caruncho, H. J.
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