Endothelin-induced myoplasmic Ca2+ responses and tyrosine phosphorylation in coronary smooth muscle

Dexter L. Lee, Michael Sturek

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

This study investigated the role of tyrosine phosphorylation and source of Ca2+ in prolonged endothelin-1 (ET-1)-induced potentiation of myoplasmic free Ca2+ ([Ca2+]m) responses to depolarization in coronary smooth muscle cells. Fura-2 microfluorometry showed typical increases in [Ca2+]m in response to 80 mM K+ (80K) and 0.01 μM endothelin. After washout of ET-1 80K-induced [Ca2+]m increases were augmented (potentiated) 31%. Time to peak [Ca2+]m response to 80K was less after ET-1 exposure than before. ET-1 potentiation of 80K-induced [Ca2+]m responses by decreased sarcoplasmic reticulum (SR) buffering of [Ca2+]m or Ca2+-induced Ca2+ release was ruled out by lack of potentiation by 5 mM caffeine and 1 μM thapsigargin. Diltiazem abolished potentiation, providing evidence for Ca2+ influx through voltage-gated Ca2+ channels (VGCC). Genistein (30 μM) and methyl 2,5-dihydroxycinnamate (1 μM, MDHC) abolished potentiation of Ca2+ influx. Single cell phosphotyrosine measured directly by immunofluorescence was increased 95% in cells treated with ET-1 compared to control, genistein, and MDHC treated cells. ET-1 increased tyrosine phosphorylation of an 80-85 kDa protein, but not the 240 kDa α1C subunit of the VGCC. Tyrosine phosphorylation of proteins other than VGCC is necessary for prolonged potentiation by ET-1 of depolarization-induced Ca2+ influx.

Original languageEnglish (US)
Pages (from-to)18-27
Number of pages10
JournalJournal of Cardiovascular Pharmacology
Volume40
Issue number1
DOIs
StatePublished - 2002
Externally publishedYes

Fingerprint

Endothelins
Endothelin-1
Smooth Muscle
Tyrosine
Phosphorylation
Genistein
Cytophotometry
Phosphotyrosine
Thapsigargin
Diltiazem
Fura-2
Sarcoplasmic Reticulum
Caffeine
Smooth Muscle Myocytes
Fluorescent Antibody Technique
Proteins

Keywords

  • Caffeine
  • Digital imaging microscopy
  • Fura-2
  • Sarcoplasmic reticulum
  • Thapsigargin
  • Voltage-gated Ca channels

ASJC Scopus subject areas

  • Pharmacology
  • Cardiology and Cardiovascular Medicine

Cite this

Endothelin-induced myoplasmic Ca2+ responses and tyrosine phosphorylation in coronary smooth muscle. / Lee, Dexter L.; Sturek, Michael.

In: Journal of Cardiovascular Pharmacology, Vol. 40, No. 1, 2002, p. 18-27.

Research output: Contribution to journalArticle

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