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Articles in PresS, published online ahead of print May 29, 2002
Am J Physiol Cell Physiol, 10.1152/ajpcell.00048.2002
Submitted on January 30, 2002
Accepted on May 20, 2002
1 Department of Medicine, University of Florida, Gainesville, Florida, USA
2 Department of Medicine, University of Florida, Gainesville, Florida, USA; Research 151, V.A. Medical Center, Gainesville, Florida, USA
* To whom correspondence should be addressed. E-mail: zhangjl{at}medicine.ufl.edu.
We investigated whether NO upregulates a cyclic nucleotide-gated (CNG) channel and whether this contributes to sustained elevation of intracellular calcium levels [Ca++]i in porcine pulmonary artery endothelial cells (PAEC). Exposure of PAEC to a NO donor, NOC 18 (1 mM), for 18 hr increased the protein and mRNA levels of CNG channel subunit 
40 % and 50 %, respectively (p < 0.05). [Ca++]i in NO-treated cells were increased 50%, and this increase was maintained for up to 12 hr after removal of NOC-18 from medium. Extracellular calcium is required for the increase in [Ca++]i in NO-treated cells. Thapsigargin induced a rapid cytosolic calcium rise, whereas both a CNG and a non-selective cation channel blocker caused a faster decline in [Ca++]i, suggesting that capacitative calcium entry contributes to the elevated calcium levels. Antisense inhibition of CNGA2 expression attenuated the NO-induced increases in CNGA2 expression and [Ca++]i and in capacitative calcium entry. Our results demonstrate that exogenous NO upregulates CNGA2 expression and that this is associated with elevated [Ca++]i and capacitative calcium entry in porcine PAEC.
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