Cellular expression of P2Y and beta-AR receptor mRNAs and proteins in freshly isolated astrocytes and tissue sections from the CA1 region of P8-12 rat hippocampus.
Although almost all GFAP(+) cells in primary astrocyte cultures show functional beta-adrenergic (beta-AR) and metabotropic purinergic (P2Y) receptors, the fewer studies on astrocytes in situ have shown that a much smaller proportion express these same receptor-mediated activities. Here we show, by multiplex single cell RT-PCR, that 44% of freshly isolated, GFAP(+) astrocytes (FIAs) from the CA1 of P8-12 rat hippocampus always co-express beta-adrenergic receptor mRNA subtypes with metabotropic ATP receptor mRNA subtypes (P2Y1, P2Y2 or P2Y4). We also found that beta2 mRNA was the dominant beta-AR subtype expressed. P2Y1 mRNA always co-expresses with either one or two subtypes of P2U-like receptor (P2Y2 or P2Y4) mRNAs. Immunocytochemical studies showed a similar percentage of all FIAs expressed beta-AR and P2Y1 protein (54% and 52%, respectively), as for the mRNAs (46% and 65%, respectively). The staining of hippocampal sections for beta-AR or P2Y1 receptor plus GFAP shows that there are quite numerous, scattered star-shaped GFAP(+) astrocytes in the CA1 region of P9-10 rat hippocampus that stained positive for either of these receptors. These data show that astrocytes in situ express, and to a large extent likely co-express, beta-AR and P2Y receptors.
Zhu
Yingzi
Y
Center for Neuropharmacology and Neuroscience, USA.
Kimelberg
Harold K
HK
D000375:Aging; D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000378:metabolism*; D002452:Cell Count; D002478:Cells, Cultured; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D006624:Hippocampus / Q000166:cytology* / Q000254:growth & development; D007150:Immunohistochemistry / Q000379:methods; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D011943:Receptors, Adrenergic, beta / Q000096:biosynthesis*; D018048:Receptors, Purinergic P2 / Q000096:biosynthesis*; D058472:Receptors, Purinergic P2Y1; D058473:Receptors, Purinergic P2Y2; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods
D005904:Glial Fibrillary Acidic Protein; C546384:P2ry1 protein, rat; C546387:P2ry2 protein, rat; D012333:RNA, Messenger; D011943:Receptors, Adrenergic, beta; D018048:Receptors, Purinergic P2; D058472:Receptors, Purinergic P2Y1; D058473:Receptors, Purinergic P2Y2