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Calcium receptor message, expression and function decrease in differentiating keratinocytes.

Sahba Fatherazi1, Carol M Belton, Shiwei Cai

  • 1Department of Oral Biology, School of Dentistry, University of Washington, Seattle, WA 98195, USA.

Pflugers Archiv : European Journal of Physiology
|February 11, 2004
PubMed
Summary
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The calcium-sensing receptor (CaSR) is present in human gingival keratinocytes (HGKs) and plays a key role in calcium-induced differentiation. CaSR signaling initiates calcium influx, crucial for HGK differentiation.

Area of Science:

  • Cell Biology
  • Physiology
  • Oral Biology

Background:

  • The calcium-sensing receptor (CaSR) is a G protein-coupled receptor that detects changes in extracellular calcium levels.
  • Keratinocytes, the main cell type in the epidermis and oral epithelium, undergo differentiation in response to various stimuli, including calcium.
  • The role of CaSR in human gingival keratinocytes (HGKs) and its involvement in differentiation remain incompletely understood.

Purpose of the Study:

  • To investigate the expression and function of CaSR in proliferating and differentiating HGKs.
  • To elucidate the role of CaSR in mediating calcium-induced differentiation in HGKs.
  • To characterize the ion currents activated by extracellular calcium in HGKs.

Main Methods:

  • Primary human gingival keratinocytes (HGKs) were cultured under conditions of low (0.03 mM) and high (1.2 mM) extracellular calcium to induce proliferation and differentiation, respectively.

Related Experiment Videos

  • CaSR mRNA and protein expression were assessed using RT-PCR and Western blotting.
  • Whole-cell patch-clamp electrophysiology was employed to measure ion currents in response to extracellular calcium.
  • Intracellular calcium ([Ca2+]) changes were monitored using calcium imaging.

    • Main Results:

    • CaSR mRNA and protein were detected in proliferating HGKs and decreased upon differentiation induction.
    • Extracellular calcium (10 mM) activated two distinct whole-cell currents in proliferating HGKs: a transient I(CRAC)-like current and a larger, sustained current.
    • Both currents were diminished in differentiating cells, and neomycin inhibited the late current.
    • Extracellular calcium dose-dependently increased intracellular calcium in proliferating HGKs, with the induced currents supporting calcium influx.
    • La3+ and 2-APB inhibited both the currents and intracellular calcium changes, suggesting CaSR involvement.

    Conclusions:

    • CaSR is expressed in HGKs and its expression is modulated during differentiation.
    • CaSR signaling is activated by extracellular calcium and mediates calcium influx through distinct ion currents.
    • CaSR-dependent calcium signaling plays a significant role in initiating calcium-induced differentiation in HGKs.