The functional expression of extracellular calcium-sensing receptor in rat pulmonary artery smooth muscle cells

J Biomed Sci. 2011 Feb 11;18(1):16. doi: 10.1186/1423-0127-18-16.

Abstract

Background: The extracellular calcium-sensing receptor (CaSR) belongs to family C of the G protein coupled receptors. Whether the CaSR is expressed in the pulmonary artery (PA) is unknown.

Methods: The expression and distribution of CaSR were detected by RT-PCR, Western blotting and immunofluorescence. PA tension was detected by the pulmonary arterial ring technique, and the intracellular calcium concentration ([Ca2+]i) was detected by a laser-scanning confocal microscope.

Results: The expressions of CaSR mRNA and protein were found in both rat pulmonary artery smooth muscle cells (PASMCs) and PAs. Increased levels of [Ca2+]o (extracellular calcium concentration) or Gd3+ (an agonist of CaSR) induced an increase of [Ca2+]i and PAs constriction in a concentration-dependent manner. In addition, the above-mentioned effects of Ca2+ and Gd3+ were inhibited by U73122 (specific inhibitor of PLC), 2-APB (specific antagonist of IP3 receptor), and thapsigargin (blocker of sarcoplasmic reticulum calcium ATPase).

Conclusions: CaSR is expressed in rat PASMCs, and is involved in regulation of PA tension by increasing [Ca2+]i through G-PLC-IP3 pathway.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Blotting, Western
  • Boron Compounds / pharmacology
  • Calcium / pharmacology
  • Calcium Channel Blockers / pharmacology
  • Enzyme Inhibitors / pharmacology
  • Estrenes / pharmacology
  • Fluorescent Antibody Technique
  • In Vitro Techniques
  • Male
  • Molecular Sequence Data
  • Myocytes, Smooth Muscle / cytology
  • Myocytes, Smooth Muscle / metabolism
  • Pulmonary Artery / cytology
  • Pulmonary Artery / metabolism*
  • Pyrrolidinones / pharmacology
  • RNA, Messenger / genetics
  • Rats
  • Rats, Wistar
  • Receptors, Calcium-Sensing / genetics*
  • Receptors, Calcium-Sensing / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Thapsigargin / pharmacology
  • Type C Phospholipases / antagonists & inhibitors

Substances

  • Boron Compounds
  • Calcium Channel Blockers
  • Enzyme Inhibitors
  • Estrenes
  • Pyrrolidinones
  • RNA, Messenger
  • Receptors, Calcium-Sensing
  • 1-(6-((3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione
  • Thapsigargin
  • 2-aminoethoxydiphenyl borate
  • Type C Phospholipases
  • Calcium