Abstract
Stromal interaction molecules (STIM1, 2) are acting as sensors for Ca2+ in intracellular stores and activate Orai channels at the plasma membrane for store-operated Ca2+ entry (SOCE), while classical transient receptor potential (TRPC) channel mediate receptor-operated Ca2+ entry (ROCE). Several reports, however, indicate a role for TRPC in SOCE in certain cell types. Here, we analyzed Ca2+ influx and cell function in TRPC1/6-deficient (TRPC1/6−/−) and STIM1/2- deficient (STIM1/2ΔpmLF) primary murine lung fibroblasts (pmLF). As expected, SOCE was decreased in STIM1/2- deficient pmLF and ROCE was decreased in TRPC1/6−/− pmLF compared to control cells. By contrast, SOCE was not significantly different in TRPC1/6−/− pmLF and ROCE was similar in STIM1/2-deficient pmLF compared to Wt cells. Most interestingly, cell proliferation, migration and nuclear localization of nuclear factor of activated T-cells (NFATc1 and c3) were decreased after ablation of STIM1/2 proteins in pmLF. In conclusion, TRPC1/6 channels are not involved in SOCE and STIM1/2 deficiency resulted in decreased cell proliferation and migration in pmLF.
Highlights
Stromal interaction molecules (STIM1, 2) are acting as sensors for Ca2+ in intracellular stores and activate Orai channels at the plasma membrane for store-operated Ca2+ entry (SOCE), while classical transient receptor potential (TRPC) channel mediate receptor-operated Ca2+ entry (ROCE)
To examine the effect of STIM deficiency in primary murine lung fibroblasts, we isolated these cells from mice with loxP flanked Stim[1] and Stim[2] genes[26] and infected them with recombinant lentiviruses expressing Cre-recombinase
DNA synthesis as a measure of cell proliferation was significantly reduced in STIM1/2- deficient fibroblasts in comparison to Wt cells infected with recombinant lentiviruses expressing Cre recombinase (Fig. 6b,c)
Summary
Stromal interaction molecules (STIM1, 2) are acting as sensors for Ca2+ in intracellular stores and activate Orai channels at the plasma membrane for store-operated Ca2+ entry (SOCE), while classical transient receptor potential (TRPC) channel mediate receptor-operated Ca2+ entry (ROCE). We set out to study the role of SOCE in primary murine lung fibroblasts (pmLF) using TRPC1/6and STIM1/2-deficient fibroblasts in comparison to Wt control cells. To examine the effect of STIM deficiency in primary murine lung fibroblasts (pmLF), we isolated these cells from mice with loxP flanked Stim[1] and Stim[2] genes[26] and infected them with recombinant lentiviruses expressing Cre-recombinase.
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