Cloning and functional expression of the bovine GABA(C) rho2 subunit. Molecular evidence of a widespread distribution in the CNS.
GABA(C) receptors were first described as a non-desensitizing, bicuculline- and baclofen-insensitive component in Xenopus oocytes expressing bovine retina mRNA. However, the expression, tissue distribution and functional properties of GABA(C) receptors from other areas of the CNS still remain controversial. In previous experiments, the injection of rat cerebellum mRNA into Xenopus oocytes induced the expression of receptors that generated currents with both GABA(A) and GABA(C) characteristics; the latter component apparently being given by the rho2 subunit, suggesting the expression of GABA(C) receptors in the CNS and the formation of homooligomeric receptors. In this study, using RT-PCR, we found that the rho1 and rho2 subunits are widely expressed in the CNS including areas where they have not been previously described such as the bulb, pons and the caudate nucleus. To determine if the GABA(C) component of the GABA-currents elicited by oocytes expressing cerebellum mRNA was caused by activation of homomeric GABA rho2 receptors, we cloned the corresponding cDNA and expressed it in Xenopus oocytes. It was found that oocytes injected with rho2 cDNA, efficiently formed GABA-gated homooligomeric receptors. The GABA-dose-current response gave an EC50=1.19muM and the currents were resistant to bicuculline and reversibly antagonized by the specific GABA(C) receptor antagonist TPMPA. Altogether, our results indicate a widespread distribution of both rho1 and rho2 subunits in the bovine CNS and show further that the rho2 subunit cDNA isolated from cerebellum, forms fully functional receptors when expressed in Xenopus oocytes.
López-Chávez
Ariel
A
Instituto de Neurobiología, Universidad Nacional Autónoma de México-Campus Juriquilla, AP1-1141 Querétaro, Mexico.
Miledi
Ricardo
R
Martínez-Torres
Ataúlfo
A
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000378:metabolism*; D002417:Cattle; D003001:Cloning, Molecular; D017403:In Situ Hybridization; D008969:Molecular Sequence Data; D009865:Oocytes; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism; D018079:Receptors, GABA / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014981:Xenopus
C087200:GABA-C receptor; D021122:Protein Subunits; D018079:Receptors, GABA