Article

Role of Trpc channels, Stim1 and Orai1 in PGF(2α)-induced calcium signaling in NRK fibroblasts.

Department of Cell Biology, Radboud University Nijmegen, AJ Nijmegen, The Netherlands.
Cell calcium (impact factor: 4.29). 11/2011; 51(1):12-21. DOI:10.1016/j.ceca.2011.10.001 pp.12-21
Source: PubMed

ABSTRACT Normal rat kidney (NRK) fibroblasts exhibit growth-dependent changes in electrophysiological properties and intracellular calcium dynamics. The transition from a quiescent state to a density-arrested state results in altered calcium entry characteristics. This coincides with modulation of the expression of the genes encoding the calcium channels Trpc1, Trpc6 and Orai1, and of the intracellular calcium sensor Stim1. In the present study we have used gene selective short hairpin (sh) RNAs against these various genes to investigate their role in (a) capacitative store-operated calcium entry (SOCE); (b) non-capacitative OAG-induced receptor-operated calcium entry (ROCE); and (c) prostaglandin F(2α) (PGF(2α))-induced Ca(2+)-oscillations in NRK fibroblasts. Intracellular calcium measurements revealed that knockdown of the genes encoding Trpc1, Orai1 and Stim1 each caused a significant reduction of SOCE in NRK cells, whereas knockdown of the gene encoding Trpc6 reduced only the OAG-induced ROCE. Furthermore, our data show that knockdown of the genes encoding Trpc1, Orai1 and Stim1, but not Trpc6, substantially reduced the frequency (up to 60%) of PGF(2α)-induced Ca(2+) oscillations in NRK cells. These results indicate that in NRK cells distinct calcium channels control the processes of SOCE, ROCE and PGF(2α)-induced Ca(2+) oscillations.

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Keywords

calcium channels Trpc1
 
calcium entry characteristics
 
coincides
 
density-arrested state results
 
electrophysiological properties
 
gene encoding Trpc6
 
gene selective short hairpin
 
genes encoding
 
genes encoding Trpc1
 
intracellular calcium dynamics
 
Intracellular calcium measurements
 
intracellular calcium sensor Stim1
 
modulation
 
Normal rat kidney
 
NRK cells
 
NRK cells distinct calcium channels control
 
NRK fibroblasts
 
PGF(2α))-induced Ca(2+)-oscillations
 
processes
 
various genes