Asymmetrical, agonist-induced fluctuations in local extracellular [Ca(2+)] in intact polarized epithelia

EMBO J. 2001 Nov 15;20(22):6316-26. doi: 10.1093/emboj/20.22.6316.

Abstract

We recently proposed that extracellular Ca(2+) ions participate in a novel form of intercellular communication involving the extracellular Ca(2+)-sensing receptor (CaR). Here, using Ca(2+)-selective microelectrodes, we directly measured the profile of agonist-induced [Ca(2+)]ext changes in restricted domains near the basolateral or luminal membranes of polarized gastric acid-secreting cells. The Ca(2+)-mobilizing agonist carbachol elicited a transient, La(3+)-sensitive decrease in basolateral [Ca(2+)] (average approximately 250 microM, but as large as 530 microM). Conversely, carbachol evoked an HgCl2-sensitive increase in [Ca(2+)] (average approximately 400 microM, but as large as 520 microM) in the lumen of single gastric glands. Both responses were significantly reduced by pre-treatment with sarco-endoplasmic reticulum Ca(2+) ATPase (SERCA) pump inhibitors or with the intracellular Ca(2+) chelator BAPTA-AM. Immunofluorescence experiments demonstrated an asymmetric localization of plasma membrane Ca(2+) ATPase (PMCA), which appeared to be partially co-localized with CaR and the gastric H(+)/K(+)-ATPase in the apical membrane of the acid-secreting cells. Our data indicate that agonist stimulation results in local fluctuations in [Ca(2+)]ext that would be sufficient to modulate the activity of the CaR on neighboring cells.

Publication types

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

MeSH terms

  • Adenosine Triphosphatases / metabolism
  • Animals
  • Anti-Infective Agents, Local / pharmacology
  • Calcium / agonists*
  • Calcium / metabolism*
  • Calcium-Transporting ATPases / antagonists & inhibitors
  • Carbachol / pharmacology
  • Cell Membrane / enzymology
  • Chelating Agents / pharmacology
  • Coloring Agents / pharmacology
  • Cytoplasm / metabolism
  • Egtazic Acid / analogs & derivatives*
  • Egtazic Acid / pharmacology
  • Endoplasmic Reticulum / metabolism
  • Epithelial Cells / metabolism*
  • Epithelium / metabolism
  • Fura-2 / pharmacology
  • Gastric Mucosa / metabolism
  • Immunohistochemistry
  • Lanthanum / pharmacology
  • Mercuric Chloride / pharmacology
  • Microscopy, Fluorescence
  • Models, Biological
  • Protein Binding
  • Protein Structure, Tertiary
  • Ranidae
  • Sarcoplasmic Reticulum / metabolism
  • Sarcoplasmic Reticulum Calcium-Transporting ATPases
  • Signal Transduction

Substances

  • Anti-Infective Agents, Local
  • Chelating Agents
  • Coloring Agents
  • 1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester
  • Egtazic Acid
  • Mercuric Chloride
  • Lanthanum
  • Carbachol
  • Adenosine Triphosphatases
  • Sarcoplasmic Reticulum Calcium-Transporting ATPases
  • Calcium-Transporting ATPases
  • Calcium
  • Fura-2