Apical P2XR contribute to [Ca2+]i signaling and Isc in mouse renal MCD.
We examined P2X receptor expression and distribution in the mouse collecting duct (CD) and their functional role in Ca(2+) signaling. Both P2X(1) and P2X(4) were detected by RT-PCR and Western blot. Immunohistochemistry demonstrated apical P2X(1) and P2X(4) immunoreactivity in principal cells in the outer medullary CD (OMCD) and inner medullary CD (IMCD). Luminal ATP induced an increase in Ca(2+) signaling in native medullary CD (MCD) as measured by fluorescence imaging. ATP also induced an increase in Ca(2+) signaling in MCD cells grown in primary culture but not in the presence of P2XR antagonist PPNDS. Short circuit current (I(sc)) measurement with mouse IMCD cells showed that P2XR agonist BzATP induced a larger I(sc) than did P2YR agonist UTP in the apical membrane. Our data reveal for the first time that P2X(1) and P2X(4) are cell-specific with prominent immunoreactivity in the apical area of MCD cells. The finding that P2XR blockade inhibits ATP-induced Ca(2+) signaling suggests that activation of P2XR is a key step in Ca(2+)-dependent purinergic signaling. The result that activation of P2XR produces large I(sc) indicates the necessity of P2XR in renal CD ion transport.
Li
Liuzhe
L
North Florida/South Georgia Veterans Health System, Gainesville, FL 32608-1135, USA.
Lynch
I Jeanette
IJ
Zheng
Wencui
W
Cash
Melanie N
MN
Teng
Xueling
X
Wingo
Charles S
CS
Verlander
Jill W
JW
Xia
Shen-Ling
SL
D000818:Animals; D020013:Calcium Signaling; D002478:Cells, Cultured; D005260:Female; D005786:Gene Expression Regulation; D007150:Immunohistochemistry; D007685:Kidney Tubules, Collecting / Q000378:metabolism*; D008297:Male; D051379:Mice; D012333:RNA, Messenger / Q000235:genetics; D018048:Receptors, Purinergic P2 / Q000145:classification / Q000235:genetics / Q000378:metabolism*
D012333:RNA, Messenger; D018048:Receptors, Purinergic P2