Down-regulation of inositol 1,4,5-trisphosphate receptor in cells stably expressing the constitutively active angiotensin II N111G-AT(1) receptor

Mol Endocrinol. 2004 Dec;18(12):2967-80. doi: 10.1210/me.2003-0488. Epub 2004 Aug 26.

Abstract

The diverse cellular changes brought about by the expression of a constitutively active receptor are poorly understood. QBI-human embryonic kidney 293A cells stably expressing the constitutively active N111G-AT(1) receptor (N111G cells) showed elevated levels of inositol phosphates and frequent spontaneous intracellular Ca(2+) oscillations. Interestingly, Ca(2+) transients triggered with maximal doses of angiotensin II were much weaker in N111G cells than in wild-type cells. These blunted responses were observed independently of the presence or absence of extracellular Ca(2+) and were also obtained when endogenous muscarinic and purinergic receptors were activated, revealing a heterologous desensitization process. The desensitized component of the Ca(2+) signaling cascade was neither the G protein G(q) nor phospholipase C. The intracellular Ca(2+) store of N111G cells and their mechanism of Ca(2+) entry also appeared to be intact. The most striking adaptive response of N111G cells was a down-regulation of their inositol 1,4,5-trisphosphate receptor (IP(3)R) as revealed by reduced IP(3)-induced Ca(2+) release, lowered [(3)H]IP(3) binding capacity, diminished IP(3)R immunoreactivity, and accelerated IP(3)R degradation involving the lysosomal pathway. Treatment with the inverse agonist EXP3174 reversed the desensitized phenotype of N111G cells. Down-regulation of IP(3)R represents a reversible adaptive response to protect cells against the adverse effects of constitutively active Ca(2+)-mobilizing receptors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Angiotensin II / pharmacology
  • Angiotensin II / physiology*
  • Antihypertensive Agents / pharmacology
  • Calcium / metabolism
  • Calcium Channels / genetics*
  • Calcium Channels / metabolism
  • Calcium Signaling / drug effects
  • Calcium Signaling / physiology*
  • Cell Line
  • Down-Regulation*
  • Embryo, Mammalian / cytology
  • Humans
  • Imidazoles / pharmacology
  • Inositol 1,4,5-Trisphosphate / metabolism
  • Inositol 1,4,5-Trisphosphate / physiology
  • Inositol 1,4,5-Trisphosphate Receptors
  • Kidney / cytology
  • Kidney / embryology
  • Losartan
  • RNA, Messenger / analysis
  • RNA, Messenger / metabolism
  • Receptor, Angiotensin, Type 1 / drug effects
  • Receptor, Angiotensin, Type 1 / genetics
  • Receptor, Angiotensin, Type 1 / metabolism*
  • Receptors, Cytoplasmic and Nuclear / genetics*
  • Receptors, Cytoplasmic and Nuclear / metabolism
  • Tetrazoles / pharmacology
  • Thapsigargin / pharmacology

Substances

  • Antihypertensive Agents
  • Calcium Channels
  • ITPR1 protein, human
  • Imidazoles
  • Inositol 1,4,5-Trisphosphate Receptors
  • RNA, Messenger
  • Receptor, Angiotensin, Type 1
  • Receptors, Cytoplasmic and Nuclear
  • Tetrazoles
  • Angiotensin II
  • Thapsigargin
  • Inositol 1,4,5-Trisphosphate
  • losartan carboxylic acid
  • Losartan
  • Calcium