We have used single- and double-labeling immunfluorescence and reverse transcription-polymerase chain reaction (RT-PCR) methods to examine expression of P2X receptor subtypes on microglial cells of brain in late embryonic and postnatal rat, in the N9 microglial cell line and primary cultured microglial cells. P2X1, P2X4, and P2X7 receptors were shown on microglial cells from late embryonic day 16. Almost all the microglial cells that were positive for the marker ED1, expressed P2X1 and P2X4 receptors, whereas only about 30% of the cells with ED1-immunoreactivity were found to express the P2X7 receptor. Positive cells were localized mainly in the white matter and around ventricles. From postnatal day 7, many microglial cells with P2X4 receptor-immunoreactivity were seen around the blood vessels. At postnatal day 30, microglial cells with P2X1 receptor-immunoreactivity disappeared and the cells with P2X4 receptor-immunoreactivity were mainly localized around blood vessels and lining the subarachnoid space. From postnatal day 30, the microglial cells with P2X7 receptor-immunoreactivity were found to be distributed widely in the forebrain. Cells with P2X7 receptor-immunoreactivity from P30 were not labeled by ED1, but some were labeled by isolectin B4. The expression of P2X1, P2X4, and P2X7 receptor mRNA and protein on primary cultures of rat microglial cells and on the N9 microglial cell line was demonstrated with immunocytochemistry and RT-PCR. This is the first report that the P2X1 receptor is expressed on microglial cells, at least in early development, before postnatal day 30.
(c) 2005 Wiley-Liss, Inc.