Altered protein kinase C activation of Na+/Ca2+ exchange in mesangial cells from salt-sensitive rats

Academic Article


  • The purpose of these studies was to determine whether there is a defect in protein kinase C (PKC) regulation of the Na+/Ca2+ exchanger in cultured mesangial cells (MC) from Dahl/Rapp salt-sensitive (S) and salt-resistant (R) rats. R and S MGs were cultured, grown on coverslips, and loaded with fura 2 for measurement of single cell cytosolic calcium concentration ([Ca2+];) in a microscope-based photometry system. Studies were performed in cells that were exposed to serum (serum fed) and in cells that were serum deprived for 24 h. Baseline [Ca2+]i values measured in a Ringer solution containing 150 mM NaCl were similar between R and S MGs in both serum-fed and serum-deprived groups, although baseline [Ca2+]j values were uniformly higher in the serum-deprived groups. Exchanger activity was assessed by reducing extracellular Na (Nae) from 150 to 2 mM, which resulted in movement of Na+ out of and Ca2+ into these cells (reverse-mode Na+/ Ca2+ exchange). PKC was activated in these cells with 15-min exposure to 100 nM phorbol 12-myristate 13-acetate (PMA). In the absence of PMA, the change in [Ca2+]i (A[Ca2+]j) with reduction in Nae was similar between R and S MCs in both serum-fed and serum-deprived groups, although the magnitude of A[Ca2+]j was enhanced by serum deprivation. In both serum-fed and serum-deprived groups, PMA significantly increased A[Ca2+]; in R but not S MGs. Upregulation of exchanger activity in R MCs could be abolished by prior 24-h exposure to PMA, a maneuver that downregulates PKC activity. Other studies were performed to evaluate exchanger protein expression using monoclonal and polyclonal antibodies. Immunoblots of PMA-treated cells revealed an increase in the levels of 70- and 120-kDa proteins in the crude membrane fraction of R but not S MCs, an increase which was abrogated by prior 24-h PMA pretreatment and corresponded to reduction in the 70-kDa protein in the crude cytosolic fraction. These data demonstrate that PKC enhances Na+/ Ca2+ exchange activity in MCs from R but not from S rats, suggesting that there may be a defect in the PKC-Na+/Ca2+ exchange regulation pathway in MCs of S rats. Copyright ©1999 the American Physiological Society.
  • Published In

    Pubmed Id

  • 23369786
  • Author List

  • Mashburn NA; Unlap MT; Runquist J; Alderman A; Johnson GVW; Bell PD
  • Volume

  • 276
  • Issue

  • 4 PART 2