Protein kinase C contributes to desensitization of ANG II signaling in adult rat cardiac fibroblasts

Am J Physiol Cell Physiol. 2000 Dec;279(6):C1978-85. doi: 10.1152/ajpcell.2000.279.6.C1978.

Abstract

We have studied G(q)-linked ANG II signaling [inositol phosphate (IP) accumulation, Ca(2+) mobilization] in primary cultures of rat cardiac fibroblasts (CFs) and have found that ANG II initiates a protein kinase C (PKC)-mediated negative feedback loop that rapidly terminates the ANG II response. Pharmacological inhibition of PKC by staurosporine and GF-109203X doubled IP production over that achieved in response to ANG II alone. Inhibition of PKC also led to larger Ca(2+) transients in response to ANG II, suggesting that Ca(2+) mobilization was proportional to G(q)-phospholipase C-IP(3) activity under the conditions studied. Depletion of cellular PKC by overnight treatment with phorbol 12-myristate 13-acetate (PMA) similarly augmented ANG II-induced IP production. Acute activation of PKC by PMA halved IP formation, with an EC(50) approximately 1 nM; 4alpha-PMA was inactive. Time course data demonstrated that ANG II-mediated IP production fully desensitized within 30 s; PKC inhibition reduced the rate and extent of this desensitization. In cells desensitized to ANG II, a purinergic agonist still mobilized intracellular Ca(2+), indicating that desensitization was homologous. The ANG II-induced Ca(2+) signal was fully resensitized within 30 min. The data demonstrate that a large portion of the IP-Ca(2+) responses of rat CFs to ANG II are short-lived because of rapid, PKC-mediated desensitization.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Age Factors
  • Angiotensin II / pharmacology*
  • Animals
  • Buffers
  • Calcium / metabolism
  • Cells, Cultured
  • Chelating Agents / pharmacology
  • Egtazic Acid / analogs & derivatives*
  • Egtazic Acid / pharmacology
  • Fibroblasts / chemistry
  • Fibroblasts / cytology
  • Fibroblasts / enzymology
  • Inositol Phosphates / metabolism
  • Male
  • Myocardium / chemistry
  • Myocardium / cytology*
  • Myocardium / enzymology*
  • Protein Kinase C / antagonists & inhibitors
  • Protein Kinase C / metabolism*
  • Rats
  • Rats, Sprague-Dawley
  • Receptor, Angiotensin, Type 1
  • Receptor, Angiotensin, Type 2
  • Receptors, Angiotensin / metabolism
  • Receptors, Purinergic / metabolism
  • Signal Transduction / drug effects
  • Signal Transduction / physiology*
  • Uridine Triphosphate / pharmacology
  • Vasoconstrictor Agents / pharmacology*

Substances

  • Buffers
  • Chelating Agents
  • Inositol Phosphates
  • Receptor, Angiotensin, Type 1
  • Receptor, Angiotensin, Type 2
  • Receptors, Angiotensin
  • Receptors, Purinergic
  • Vasoconstrictor Agents
  • Angiotensin II
  • Egtazic Acid
  • Protein Kinase C
  • 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid
  • Calcium
  • Uridine Triphosphate