Store-operated Ca 2+ entry in primary murine lung fibroblasts is independent of classical transient receptor potential (TRPC) channels and contributes to cell migration

Sci Rep. 2020 Apr 22;10(1):6812. doi: 10.1038/s41598-020-63677-2.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calcium / metabolism*
  • Cell Movement* / drug effects
  • Cell Nucleus / drug effects
  • Cell Nucleus / metabolism
  • Cell Proliferation / drug effects
  • Cell Survival / drug effects
  • Cells, Cultured
  • DNA / biosynthesis
  • Endothelin-1 / pharmacology
  • Exons / genetics
  • Fibroblasts / cytology*
  • Fibroblasts / metabolism*
  • Integrases / metabolism
  • Lung / cytology*
  • Mice, Inbred C57BL
  • NFATC Transcription Factors / metabolism
  • ORAI1 Protein / genetics
  • ORAI1 Protein / metabolism
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Stromal Interaction Molecule 1 / deficiency
  • Stromal Interaction Molecule 1 / genetics
  • Stromal Interaction Molecule 1 / metabolism
  • Stromal Interaction Molecule 2 / deficiency
  • Stromal Interaction Molecule 2 / genetics
  • Stromal Interaction Molecule 2 / metabolism
  • Thapsigargin / pharmacology
  • Transient Receptor Potential Channels / metabolism*

Substances

  • Endothelin-1
  • NFATC Transcription Factors
  • ORAI1 Protein
  • RNA, Messenger
  • Stim2 protein, mouse
  • Stromal Interaction Molecule 1
  • Stromal Interaction Molecule 2
  • Transient Receptor Potential Channels
  • Thapsigargin
  • DNA
  • Cre recombinase
  • Integrases
  • Calcium