Functional expression of metabotropic GABAB receptors in primary cultures of astrocytes from rat cerebral cortex.
GABA(B) receptor subunits are widely expressed on neurons throughout the central nervous system (CNS), at both pre- and postsynaptic sites, where they mediate the late and slow component of the inhibitory response to the major inhibitory neurotransmitter GABA. Recently, GABA(B) receptors have been reported to be expressed in astrocytes and microglia in the rat CNS by immunocytochemistry. However, there are few reports available for the functional characterization of GABA(B) receptors on astrocytes. In the present study, we therefore investigated the functional expression and characteristics of GABA(B) receptors in primary cultures of astrocytes from rat cerebral cortex. In the presence of 10 microM GTP, forskolin concentration-dependently increased adenylylcyclase (AC) activity in membranes prepared from rat astrocytes. The selective GABA(B) agonist (R)-baclofen concentration-dependently reduced forskolin-stimulated AC activity in the presence of 10 microM GTP. This effect was reversed by the selective GABA(B) antagonists, CGP-55845 and CGP-54626, and was completely abolished by treatment of astrocytic membranes with pertussis toxin. In addition, RT-PCR, Western blotting, and immunocytochemistry clearly showed that metabotropic GABA(B) receptor isoforms (GABA(B)R1 and GABA(B)R2) are expressed in rat cerebrocortical astrocytes. Taken collectively, these results demonstrate that functionally active metabotropic GABA(B) receptors are expressed in rat cerebrocortical astrocytes.
Oka
Michiko
M
Department of Biochemical Pharmacology, Kyoto Pharmaceutical University, Misasagi, Kyoto, Japan.
Wada
Miyuki
M
Wu
Qiang
Q
Yamamoto
Akira
A
Fujita
Takuya
T
D000067956:Adenylyl Cyclase Inhibitors; D000262:Adenylyl Cyclases / Q000235:genetics / Q000378:metabolism; D000818:Animals; D001253:Astrocytes / Q000187:drug effects / Q000378:metabolism*; D001418:Baclofen / Q000494:pharmacology; D002462:Cell Membrane / Q000187:drug effects / Q000201:enzymology; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology* / Q000187:drug effects; D005576:Colforsin / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D006160:Guanosine Triphosphate / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D018080:Receptors, GABA-B / Q000378:metabolism*
D000067956:Adenylyl Cyclase Inhibitors; D012333:RNA, Messenger; D018080:Receptors, GABA-B; D005576:Colforsin; D006160:Guanosine Triphosphate; D000262:Adenylyl Cyclases; D001418:Baclofen