Chicken GABA(A) receptor beta4 subunits form robust homomeric GABA-gated channels in Xenopus oocytes.
Chicken GABA(A) receptor beta4L and beta4S subunits were expressed in Xenopus oocytes by cRNA injection. Oocytes expressing either beta4 subunit alone or in combination with the chicken alpha1 subunit were studied using the two-electrode voltage-clamp technique. Both the beta4L and beta4S subunits form homomeric GABA-gated Cl- channels with similar efficiencies. In comparison, oocytes expressing either the chicken alpha1 or beta2S polypeptide show no or barely detectable GABA responses, as reported by others for most single-subunit vertebrate GABA(A) receptors. The GABA-gated currents due to the beta4L-subunit homomer were not affected by the presence of actinomycin D during cRNA expression, indicating that nascent oocyte polypeptides are not required for channel formation. The homomeric beta4L-subunit receptors show high affinity for GABA with an EC50 value of 4.3 +/- 0.4 microM and a Hill coefficient of 1.1 +/- 0.1 (n = 6). In response to GABA application at the EC25 value, currents elicited from the beta4L-subunit receptor are enhanced by 50 microM pentobarbital (110 +/- 10%, n = 3) and 10 microM loreclezole (60 +/- 3%, n = 3), inhibited by 10 microM picrotoxinin (93 +/- 3%, n = 3), but not affected by 1 microM diazepam. These properties are similar to those found for oocytes expressing heteromeric chicken alpha1beta4L and alpha1beta2S receptors. Since the beta subunits of GABA(A) receptors provide essential determinants for receptor assembly and subcellular localization, homomeric beta4-subunit receptors are a useful model system for further study of the structure and function of GABA(A) receptors.
Liu
S C
SC
Verna and Marrs McLean Department of Biochemistry, Baylor College of Medicine, Houston, TX 77030, USA.
Parent
L
L
Harvey
R J
RJ
Darlison
M G
MG
Barnes
E M
EM
D000494:Allosteric Regulation; D000818:Animals; D002645:Chickens; D004305:Dose-Response Relationship, Drug; D015640:Ion Channel Gating; D007473:Ion Channels / Q000378:metabolism; D009865:Oocytes / Q000378:metabolism*; D018408:Patch-Clamp Techniques; D011963:Receptors, GABA-A / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D014981:Xenopus; D005680:gamma-Aminobutyric Acid / Q000378:metabolism
D007473:Ion Channels; D011963:Receptors, GABA-A; D011994:Recombinant Proteins; D005680:gamma-Aminobutyric Acid