Clustered and non-clustered GABAA receptors in cultured hippocampal neurons.
In cultured hippocampal neurons, gamma2 subunit-containing GABA(A) Rs form large postsynaptic clusters at GABAergic synapses and small clusters outside GABAergic synapses. We now show that a pool of non-clustered gamma2 subunit-containing GABA(A) Rs are also present at the cell surface. We also demonstrate that myc- or EGFP-tagged gamma2, alpha2, beta3 or alpha1 subunits expressed in these neurons assemble with endogenous subunits, forming GABA(A) Rs that target large postsynaptic clusters, small clusters outside GABAergic synapses or a pool of non-clustered surface GABA(A) Rs. In contrast, myc- or EGFP-tagged delta subunits only form non-clustered GABA(A) Rs, which can be induced to form clusters by antibody capping. A myc-tagged chimeric gamma2 subunit possessing the large intracellular loop (IL) of the delta-subunit IL (myc gamma2S/delta-IL) assembled into GABA(A) Rs, but it did not form clusters, therefore behaving like the delta subunit. Thus, the large intracellular loops of gamma2 and delta play an important role in determining the synaptic clustering/non-clustering capacity of the GABA(A) Rs.
Christie
S B
SB
Department of Physiology and Neurobiology, University of Connecticut, 3107 Horsebarn Hill Rd., U-4156, Storrs, CT 06269, USA.
Li
R-W
RW
Miralles
C P
CP
Yang
B -Y
B-
De Blas
A L
AL
D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D003001:Cloning, Molecular; D017931:DNA Primers; D006624:Hippocampus / Q000502:physiology*; D008969:Molecular Sequence Data; D009474:Neurons / Q000502:physiology*; D051381:Rats; D011963:Receptors, GABA-A / Q000502:physiology*; D013569:Synapses / Q000502:physiology
D017931:DNA Primers; D011963:Receptors, GABA-A