Selective activation of excitation-contraction coupling pathways by ET(A) and ET(B) receptors in guinea-pig tracheal smooth muscle

Br J Pharmacol. 1999 Feb;126(4):893-902. doi: 10.1038/sj.bjp.0702365.

Abstract

1. Signalling events responsible for endothelin(A) (ET(A)) and ET(B) receptor-induced contraction were examined in epithelium-denuded guinea-pig tracheal smooth muscle strips. Selective stimulation of each subtype was achieved by a combination of ET-1 (100 nM) and ET(A) and ET(B) receptor-selective antagonists, BQ-123 (10 microM) and BQ-788 (3 microM), respectively. 2. Both ET(A) and ET(B) receptors induced long-lasting contraction that was totally dependent on Ca2+ influx. Stimulation of ET(A) receptor induced both transient and sustained (Ca2+)i increases whereas that of ET(B) receptor induced only a sustained increase. Suppression of the transient (Ca2+)i increase by U73122 (3 microM) did not affect the ET(A)-induced sustained (Ca2+)i increase and tension development. Stimulation of ET(A) receptor, but not ET(B), induced phosphoinositide breakdown and protein kinase C (PKC). The activated PKC contributed to the contraction by increasing the Ca2+ sensitivity of the contractile apparatus. 3. Thus, ET(A) receptor is coupled both with phospholipase C/Ca2+/PKC signalling and Ca2+ influx pathways whereas ET(B) receptor was coupled only with the latter. 4. Stimulation of ET(B) receptor, but not ET(A), caused membrane depolarization measured with a fluorescent indicator, bis-(1,3 dibutylbarbituric acid)-trimethine oxonol. Both nifedipine (1 microM) and verapamil (10 microM) abolished ET(B)-induced Ca2+ influx and contraction, while they barely affected ET(A)-induced responses. 5. Therefore, the Ca2+ influx pathways activated by each subtype appeared to be completely different; ET(A) and ET(B) receptors opens voltage-independent Ca2+ channels and L-type voltage-dependent Ca2+ channels, respectively.

MeSH terms

  • Animals
  • Calcium / metabolism
  • Endothelin-1 / pharmacology
  • Guinea Pigs
  • In Vitro Techniques
  • Male
  • Muscle Contraction / drug effects
  • Muscle, Smooth / physiology*
  • Protein Kinase C / physiology
  • Receptor, Endothelin A
  • Receptor, Endothelin B
  • Receptors, Endothelin / physiology*
  • Trachea / physiology*
  • Type C Phospholipases / physiology

Substances

  • Endothelin-1
  • Receptor, Endothelin A
  • Receptor, Endothelin B
  • Receptors, Endothelin
  • Protein Kinase C
  • Type C Phospholipases
  • Calcium