Ectopically expressed gamma-aminobutyric acid receptor B is functionally down-regulated in isolated lipid raft-enriched membranes.

Lipid raft domains have attracted much recent attention as platforms for plasma membrane signalling complexes. In particular, evidence is emerging that shows them to be key regulators of G protein coupled receptor function. The G protein coupled gamma-aminobutyric acid receptor B (GABA(B) receptor) co-isolates with lipid raft domains from rat brain cerebellum. In the present study, we show that the GABA(B1a,2) receptor was also present in lipid raft domains when expressed ectopically in a Chinese hamster ovary cell line. Lipid raft-associated receptor was functionally active, displaying a concentration-dependent increase in GTPgammaS binding in response to the receptor agonist GABA. Compared with whole cell membranes, lipid raft-associated receptor displayed an increased EC(50) and a reduced magnitude of response to GABA. We conclude that lipid raft association is an intrinsic property of the GABA(B1a,2) receptor and is not cell-type specific. In addition, localisation to lipid raft domains may provide a mechanism to inhibit receptor function.

GABAB receptor subunits, R1 and R2, in brainstem catecholamine and serotonin neurons.

GABA(B) receptors have been implicated in the GABAergic modulation of catecholaminergic and serotonergic pathways in the central nervous system. The GABA(B) receptor may require two subunits, GABA(B)R1 and GABA(B)R2, for functional activity. Using dual immunofluorescent labelling on adjacent cryostat sections, we investigated the presence of immunoreactivity for the GABA(B)R1 and GABA(B)R2 subunits in brainstem catecholamine (tyrosine hydroxylase-immunoreactive) and serotonin (tryptophan hydroxylase-immunoreactive) neurons. All neurons (>98%) examined in catecholamine groups A1, A2, A5, A6, C1, and serotonin groups B1-3 and B6-8 were immunoreactive for the GABA(B)R1 subunit. All A5 and A6 neurons (>97%) and at least 86% of A1, A2, C1, B2, B3, B7 and B8 neurons examined were GABA(B)R2-immunoreactive. The proportion of neurons with immunoreactivity for the GABA(B)R2 subunit varied between 0% and 99% for B1 neurons, and between 35% and 93% for B6 neurons. Statistical analysis showed that similar proportions of sampled neurons were immunoreactive for GABA(B)R1 and GABA(B)R2 in the A1, A5, A6, C1, B2 and B7 cell groups, whereas a smaller proportion of A2, B1, B3, B6 and B8 neurons were GABA(B)R2-immunoreactive than GABA(B)R1-immunoreactive. In general, our results suggest that GABA(B)R1 and GABA(B)R2 co-exist in the great majority of brainstem catecholamine and serotonin neurons. In the neurons that lack GABA(B)R2, the GABA(B)R1 subunit may act alone or with another protein.