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Related Experiment Videos

Does nitric oxide regulate capacitative Ca influx in HEK 293 cells?

G Bischof1, T F Serwold, T E Machen

  • 1Department of Molecular and Cell Biology, University of California, Berkeley 94720-3200, USA.

Cell Calcium
|February 1, 1997
PubMed
Summary

Capacitative calcium (Ca) entry in human embryonic kidney 293 cells is stimulated by calcium store depletion but not regulated by nitric oxide synthase (NOS) and cyclic guanosine monophosphate (cGMP). This contrasts with pancreatic and colonic cells, suggesting varied regulation mechanisms.

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Area of Science:

  • Cellular Physiology
  • Molecular Biology
  • Signaling Pathways

Background:

  • Capacitative calcium influx is proposed to be regulated by nitric oxide synthase (NOS) and cyclic guanosine monophosphate (cGMP) in pancreatic acinar and HT-29 colonic cells.
  • Nitric oxide (NO) influences cGMP levels via guanylate cyclase, a key step in cellular signaling.
  • Understanding the role of NO/cGMP in calcium entry is crucial for various physiological processes.

Purpose of the Study:

  • To investigate whether capacitative calcium (Ca) entry in human embryonic kidney 293 cells is regulated by nitric oxide synthase (NOS) and cyclic guanosine monophosphate (cGMP).
  • To compare calcium entry regulation mechanisms between non-excitable 293 cells and excitable epithelial cells.

Main Methods:

  • Measured Ca (or Ba) entry into wild-type and neuronal NOS-transfected human embryonic kidney 293 cells using digital imaging microscopy and Fura-2 fluorescence.

Related Experiment Videos

  • Stimulated Ca entry using cyclopiazonic acid (CPA) to deplete internal Ca stores.
  • Assessed cyclic guanosine monophosphate (cGMP) levels using an EIA method and modulated them with NOS inhibitor N-nitro-L-arginine (L-NA) and guanylate cyclase inhibitor LY83583.

    • Main Results:

    • Capacitative Ca entry, stimulated by CPA, was observed in both 293 and 293/NOS cells but was unaffected by the NOS inhibitor L-NA.
    • cGMP levels were significantly higher in 293/NOS cells than in 293 cells and were reduced by L-NA and LY83583.
    • No correlation was found between cGMP levels and the rate of Ca entry across different experimental conditions.

    Conclusions:

    • Capacitative Ca entry in human embryonic kidney 293 cells is primarily regulated by Ca store depletion, independent of NO/cGMP signaling.
    • This finding contrasts with mechanisms observed in pancreatic and colonic cells, indicating cell-specific regulation of calcium entry.
    • The study suggests that NO and cGMP do not universally regulate capacitative calcium entry in all non-excitable cells.