Co-assembly of GABA rho subunits with the GABA(A) receptor gamma(2) subunit cloned from white perch retina.
Although it is well established that GABA(C) receptors are comprised in part of GABA rho subunits, the exact subunit composition of neuronal GABA(C) receptors is yet to be determined. A detailed comparison of GABA(C)-mediated neuronal responses elicited from retinal neurons with those obtained from receptors formed by GABA rho subunits revealed a number of significant differences both in the kinetics and the pharmacology of the responses. Our previous studies indicated that the human GABA(A) receptor gamma(2) subunit could co-assemble with one (rho(1B)) of the white perch GABA rho subunits to form a heterooligomeric receptor with properties that resembled those of the GABA(C) receptors on white perch bipolar cells. In this study, we cloned the white perch gamma(2) subunit, and investigated its co-assembly with four white perch GABA rho subunits. When expressed in Xenopus oocytes, perch gamma(2) and rho(1B) subunits form heterooligomeric receptors with distinct properties: the GABA-elicited responses have fast kinetics and are sensitive to pentobarbital modulation. The enhancement of GABA-elicited responses by pentobarbital on the heterooligomeric receptors could be eliminated by a single mutation in the third transmembrane domain of the gamma(2) subunit, indicating that pentobarbital sensitivity is mediated by the incorporated gamma(2) subunit. On the other hand, co-expression of the perch gamma(2) subunit with the other perch GABA rho subunits produced no detectable changes in the kinetics of GABA-elicited response or the sensitivity to pentobarbital modulation. These results suggest that the gamma(2) subunit can co-assemble only with one (rho(1B)), but not with other white perch GABA rho subunits.
Qian
Haohua
H
Department of Ophthalmology and Visual Sciences, University of Illinois at Chicago, 1855 West Taylor Street, Chicago, IL 60612, USA. hquian@uic.edu
Pan
Yi
Y
D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D018757:GABA Modulators / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects; D008969:Molecular Sequence Data; D009865:Oocytes / Q000502:physiology; D010424:Pentobarbital / Q000494:pharmacology; D010471:Perches; D017434:Protein Structure, Tertiary; D011963:Receptors, GABA-A / Q000235:genetics* / Q000378:metabolism*; D018080:Receptors, GABA-B / Q000235:genetics* / Q000378:metabolism*; D012160:Retina / Q000502:physiology*; D014981:Xenopus
D018757:GABA Modulators; D011963:Receptors, GABA-A; D018080:Receptors, GABA-B; D010424:Pentobarbital