Effects of ryanodine on acetylcholine-induced Ca 2+ mobilization in single smooth muscle cells of the porcine coronary artery

Hideaki Katsuyama, Shinichi Ito, Take Itoh, Hirosi Kuriyama

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

To study the essential features of acetylcholine (ACh)-and caffeine-sensitive cellular Ca 2+ storage sites in single vascular smooth muscle cells of the porcine coronary artery, the effects of ryanodine on both ACh- and caffeine-induced Ca 2+ mobilization were investigated by measuring intracellular Ca 2+ concentration ([Ca 2+ ] i ) using Fura 2 in Ca 2+ -containing or Ca 2+ -free solution. The resting [Ca 2+ ] i of the cells was 122 nM in normal physiological solution and no spontaneous activity was observed. In a solution containing 2.6 mM Ca 2+ , 10 μM ACh or 128 mM K + produced a phasic, followed by a tonic, increase in [Ca 2+ ] i but 20 mM caffeine produced only a phasic increase. In Ca 2+ -free solution containing 0.5 mM ethylenebis(oxonitrilo)tetraacetate (EGTA), the resting [Ca 2+ ] i rapidly decreased to 102 nM within 5 min, and 10 μM ACh or 20 mM caffeine (but not 128 mM K + ) transiently increased [Ca 2+ ] i . Ryanodine (50 μM) greatly inhibited the phasic increase in [Ca 2+ ] i induced by 10 μM ACh or 5 mM caffeine and increased the time to peak and to the half decay after the peak in the presence or absence of extracellular Ca 2+ . By contrast, ryanodine (50 μM) enhanced the tonic increase in [Ca 2+ ] i induced by 128 mM K + and also by 10 μM ACh in Ca 2+ -containing solution. In Ca 2+ -free solution containing 0.5 mM EGTA, ACh (10 μM) failed to increase [Ca 2+ ] i following application of 20 mM caffeine. The level of [Ca 2+ ] i induced by 20 mM caffeine was greatly reduced, but not abolished, following application of 10 μM ACh in Ca 2+ -free solution. These results suggest that both ACh and caffeine release Ca 2+ from the ryanodine-sensitive sarcoplasmic reticulum (SR) in smooth muscle cells of the porcine coronary artery. The finding that ryanodine significantly increased the resting [Ca 2+ ] i and inhibited the rate of decline of [Ca 2+ ] i following wasthout of high K + or ACh in Ca 2+ -containing solution suggests that SR may negatively regulate the resting [Ca 2+ ] i in smooth muscle cells of the porcine coronary artery.

Original languageEnglish
Pages (from-to)460-466
Number of pages7
JournalPflügers Archiv European Journal of Physiology
Volume419
Issue number5
DOIs
Publication statusPublished - Nov 1 1991

Fingerprint

Ryanodine
Acetylcholine
Smooth Muscle Myocytes
Muscle
Coronary Vessels
Caffeine
Swine
Cells
Egtazic Acid
Sarcoplasmic Reticulum
Fura-2
Vascular Smooth Muscle

All Science Journal Classification (ASJC) codes

  • Physiology
  • Clinical Biochemistry
  • Physiology (medical)

Cite this

Effects of ryanodine on acetylcholine-induced Ca 2+ mobilization in single smooth muscle cells of the porcine coronary artery . / Katsuyama, Hideaki; Ito, Shinichi; Itoh, Take; Kuriyama, Hirosi.

In: Pflügers Archiv European Journal of Physiology, Vol. 419, No. 5, 01.11.1991, p. 460-466.

Research output: Contribution to journalArticle

@article{b05f57705e8e446b924e730534d14eff,
title = "Effects of ryanodine on acetylcholine-induced Ca 2+ mobilization in single smooth muscle cells of the porcine coronary artery",
abstract = "To study the essential features of acetylcholine (ACh)-and caffeine-sensitive cellular Ca 2+ storage sites in single vascular smooth muscle cells of the porcine coronary artery, the effects of ryanodine on both ACh- and caffeine-induced Ca 2+ mobilization were investigated by measuring intracellular Ca 2+ concentration ([Ca 2+ ] i ) using Fura 2 in Ca 2+ -containing or Ca 2+ -free solution. The resting [Ca 2+ ] i of the cells was 122 nM in normal physiological solution and no spontaneous activity was observed. In a solution containing 2.6 mM Ca 2+ , 10 μM ACh or 128 mM K + produced a phasic, followed by a tonic, increase in [Ca 2+ ] i but 20 mM caffeine produced only a phasic increase. In Ca 2+ -free solution containing 0.5 mM ethylenebis(oxonitrilo)tetraacetate (EGTA), the resting [Ca 2+ ] i rapidly decreased to 102 nM within 5 min, and 10 μM ACh or 20 mM caffeine (but not 128 mM K + ) transiently increased [Ca 2+ ] i . Ryanodine (50 μM) greatly inhibited the phasic increase in [Ca 2+ ] i induced by 10 μM ACh or 5 mM caffeine and increased the time to peak and to the half decay after the peak in the presence or absence of extracellular Ca 2+ . By contrast, ryanodine (50 μM) enhanced the tonic increase in [Ca 2+ ] i induced by 128 mM K + and also by 10 μM ACh in Ca 2+ -containing solution. In Ca 2+ -free solution containing 0.5 mM EGTA, ACh (10 μM) failed to increase [Ca 2+ ] i following application of 20 mM caffeine. The level of [Ca 2+ ] i induced by 20 mM caffeine was greatly reduced, but not abolished, following application of 10 μM ACh in Ca 2+ -free solution. These results suggest that both ACh and caffeine release Ca 2+ from the ryanodine-sensitive sarcoplasmic reticulum (SR) in smooth muscle cells of the porcine coronary artery. The finding that ryanodine significantly increased the resting [Ca 2+ ] i and inhibited the rate of decline of [Ca 2+ ] i following wasthout of high K + or ACh in Ca 2+ -containing solution suggests that SR may negatively regulate the resting [Ca 2+ ] i in smooth muscle cells of the porcine coronary artery.",
author = "Hideaki Katsuyama and Shinichi Ito and Take Itoh and Hirosi Kuriyama",
year = "1991",
month = "11",
day = "1",
doi = "10.1007/BF00370789",
language = "English",
volume = "419",
pages = "460--466",
journal = "Pflugers Archiv European Journal of Physiology",
issn = "0031-6768",
publisher = "Springer Verlag",
number = "5",

}

TY - JOUR

T1 - Effects of ryanodine on acetylcholine-induced Ca 2+ mobilization in single smooth muscle cells of the porcine coronary artery

AU - Katsuyama, Hideaki

AU - Ito, Shinichi

AU - Itoh, Take

AU - Kuriyama, Hirosi

PY - 1991/11/1

Y1 - 1991/11/1

N2 - To study the essential features of acetylcholine (ACh)-and caffeine-sensitive cellular Ca 2+ storage sites in single vascular smooth muscle cells of the porcine coronary artery, the effects of ryanodine on both ACh- and caffeine-induced Ca 2+ mobilization were investigated by measuring intracellular Ca 2+ concentration ([Ca 2+ ] i ) using Fura 2 in Ca 2+ -containing or Ca 2+ -free solution. The resting [Ca 2+ ] i of the cells was 122 nM in normal physiological solution and no spontaneous activity was observed. In a solution containing 2.6 mM Ca 2+ , 10 μM ACh or 128 mM K + produced a phasic, followed by a tonic, increase in [Ca 2+ ] i but 20 mM caffeine produced only a phasic increase. In Ca 2+ -free solution containing 0.5 mM ethylenebis(oxonitrilo)tetraacetate (EGTA), the resting [Ca 2+ ] i rapidly decreased to 102 nM within 5 min, and 10 μM ACh or 20 mM caffeine (but not 128 mM K + ) transiently increased [Ca 2+ ] i . Ryanodine (50 μM) greatly inhibited the phasic increase in [Ca 2+ ] i induced by 10 μM ACh or 5 mM caffeine and increased the time to peak and to the half decay after the peak in the presence or absence of extracellular Ca 2+ . By contrast, ryanodine (50 μM) enhanced the tonic increase in [Ca 2+ ] i induced by 128 mM K + and also by 10 μM ACh in Ca 2+ -containing solution. In Ca 2+ -free solution containing 0.5 mM EGTA, ACh (10 μM) failed to increase [Ca 2+ ] i following application of 20 mM caffeine. The level of [Ca 2+ ] i induced by 20 mM caffeine was greatly reduced, but not abolished, following application of 10 μM ACh in Ca 2+ -free solution. These results suggest that both ACh and caffeine release Ca 2+ from the ryanodine-sensitive sarcoplasmic reticulum (SR) in smooth muscle cells of the porcine coronary artery. The finding that ryanodine significantly increased the resting [Ca 2+ ] i and inhibited the rate of decline of [Ca 2+ ] i following wasthout of high K + or ACh in Ca 2+ -containing solution suggests that SR may negatively regulate the resting [Ca 2+ ] i in smooth muscle cells of the porcine coronary artery.

AB - To study the essential features of acetylcholine (ACh)-and caffeine-sensitive cellular Ca 2+ storage sites in single vascular smooth muscle cells of the porcine coronary artery, the effects of ryanodine on both ACh- and caffeine-induced Ca 2+ mobilization were investigated by measuring intracellular Ca 2+ concentration ([Ca 2+ ] i ) using Fura 2 in Ca 2+ -containing or Ca 2+ -free solution. The resting [Ca 2+ ] i of the cells was 122 nM in normal physiological solution and no spontaneous activity was observed. In a solution containing 2.6 mM Ca 2+ , 10 μM ACh or 128 mM K + produced a phasic, followed by a tonic, increase in [Ca 2+ ] i but 20 mM caffeine produced only a phasic increase. In Ca 2+ -free solution containing 0.5 mM ethylenebis(oxonitrilo)tetraacetate (EGTA), the resting [Ca 2+ ] i rapidly decreased to 102 nM within 5 min, and 10 μM ACh or 20 mM caffeine (but not 128 mM K + ) transiently increased [Ca 2+ ] i . Ryanodine (50 μM) greatly inhibited the phasic increase in [Ca 2+ ] i induced by 10 μM ACh or 5 mM caffeine and increased the time to peak and to the half decay after the peak in the presence or absence of extracellular Ca 2+ . By contrast, ryanodine (50 μM) enhanced the tonic increase in [Ca 2+ ] i induced by 128 mM K + and also by 10 μM ACh in Ca 2+ -containing solution. In Ca 2+ -free solution containing 0.5 mM EGTA, ACh (10 μM) failed to increase [Ca 2+ ] i following application of 20 mM caffeine. The level of [Ca 2+ ] i induced by 20 mM caffeine was greatly reduced, but not abolished, following application of 10 μM ACh in Ca 2+ -free solution. These results suggest that both ACh and caffeine release Ca 2+ from the ryanodine-sensitive sarcoplasmic reticulum (SR) in smooth muscle cells of the porcine coronary artery. The finding that ryanodine significantly increased the resting [Ca 2+ ] i and inhibited the rate of decline of [Ca 2+ ] i following wasthout of high K + or ACh in Ca 2+ -containing solution suggests that SR may negatively regulate the resting [Ca 2+ ] i in smooth muscle cells of the porcine coronary artery.

UR - http://www.scopus.com/inward/record.url?scp=0026058833&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026058833&partnerID=8YFLogxK

U2 - 10.1007/BF00370789

DO - 10.1007/BF00370789

M3 - Article

VL - 419

SP - 460

EP - 466

JO - Pflugers Archiv European Journal of Physiology

JF - Pflugers Archiv European Journal of Physiology

SN - 0031-6768

IS - 5

ER -