γ-aminobutyric acidA ρ receptor subunits in the developing rat hippocampus

The RT-PCR approach was used to estimate the expression of gamma-aminobutyric acid (GABA)(A) rho receptor subunits in the hippocampus of neonatal and adult rats. All three p subunits were detected at postnatal day (P) 2, the rho3 subunit being expressed at an extremely low level. The rho1 and rho2 products appeared to be developmentally regulated; they were found to be more pronounced in adulthood. In another set of experiments, to correlate gene expression with receptor function, GABA(A) rho subunit mRNAs were detected with single-cell RT-PCR in CA3 pyramidal cells (from P3-P4 hippocampal slices), previously characterized with electrophysiological experiments for their bicuculline-sensitive or -insensitive responses to GABA. In 6 of 19 cells (31%), pressure application of GABA evoked at -70 mV inward currents that persisted in the presence of 100 muM bicuculline (314 +/- 129 pA). RT-PCR performed in two of these neurons revealed the presence of rho1 and rho2 subunits, the latter being present with the alpha2 subunit. A rho2 subunit was also found in 1 neuron (among 9) exhibiting a response to GABA, which was completely abolished by bicuculline. This might be due to the lack of putative accessory GABAA subunits that can coassemble with rho2 to make functional receptors. Similar experiments from 10 P15 CA3 pyramidal cells failed to reveal any rho1-3 transcripts. However, these neurons abundantly express 0 subunits. It is likely that in CA3 pyramidal cells of neonatal and adult hippocampus GABAA p subunits are present but at very low levels of expression. (C) 2002 Wiley-Liss, Inc.