Recycling and buffering of intracellular calcium in vascular smooth muscle from genetically hypertensive rats

Nancy L. Kanagy, Maria N. Ansari, Sanjay Ghosh, R. Clinton Webb

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Objective: To test the hypothesis that impaired Ca2+ recycling by the sarcoplasmic reticulum Ca-ATPase contributes to augmented force development in arteries from stroke-prone spontaneously hypertensive rats (SHRSP). Methods: Force development to caffeine (0.3-30 mmol/l) in the absence of extracellular Ca2+ was compared in aortic strips from SHRSP and Wistar-Kyoto (WKY) rats. In another protocol the strips were rinsed at the peak of contraction to caffeine (20 mmol/l) and subsequently restimulated with the alkaloid. The second response, dependent on recycled Ca2+, was used as a measure of sarcoplasmic reticulum function. A third protocol evaluated caffeine-induced contractions after Ca2+ depletion and reloading. In these latter experiments the effects of thapsigargin, an inhibitor of the sarcoplasmic reticulum Ca-ATPase, and ryanodine, an activator of sarcoplasmic reticulum Ca2+ release channels, were used to evaluate Ca2+ buffering. Finally, unidirectional 45Ca2+ influx was measured. Results: Contractions to caffeine (0.3-30 mmol/l) were larger in SHRSP aortic strips than in WKY rat strips. After a rinse at the peak of the initial response to caffeine, SHRSP segments contracted more when challenged a second time. Thapsigargin (0.3-10 mol/l) caused a concentration-dependent contraction during Ca2+ loading that was greater in SHRSP than in WKY rat strips, and a concentration-dependent inhibition of caffeine-induced contraction with similar median inhibitory concentrations in the two groups. Ryanodine did not cause contraction during Ca2+ loading, but caffeine-induced contractions were reduced after ryanodine treatment in both groups. 45Ca2+ influx was increased in SHRSP aortic segments. Conclusions: The greater force development to caffeine in SHRSP aortic strips probably reflects a greater storage of activator Ca2+ in the sarcoplasmic reticulum. On the basis of the pharmacological properties of thapsigargin and ryanodine, it appears that the larger store is caused by enhanced Ca2+ influx across the sarcolemma rather than by recycling of Ca2+ by sarcoplasmic reticulum Ca-ATPase. Experiments evaluating the secondary response to caffeine also support the interpretation that recycling of activator Ca2+ into the sarcoplasmic reticulum does not explain the augmented force development in SHRSP aortic segments.

Original languageEnglish (US)
Pages (from-to)1365-1372
Number of pages8
JournalJournal of hypertension
Volume12
Issue number12
StatePublished - Dec 1994

Fingerprint

Recycling
Caffeine
Vascular Smooth Muscle
Sarcoplasmic Reticulum
Calcium
Ryanodine
Thapsigargin
Inbred WKY Rats
Adenosine Triphosphatases
Sarcolemma
Inbred SHR Rats
Alkaloids
Arteries
Stroke
Pharmacology

Keywords

  • Caffeine
  • Hypertension
  • Ryanodine
  • Sarcoplasmic reticulum
  • Thapsigargin

ASJC Scopus subject areas

  • Internal Medicine
  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

Recycling and buffering of intracellular calcium in vascular smooth muscle from genetically hypertensive rats. / Kanagy, Nancy L.; Ansari, Maria N.; Ghosh, Sanjay; Webb, R. Clinton.

In: Journal of hypertension, Vol. 12, No. 12, 12.1994, p. 1365-1372.

Research output: Contribution to journalArticle

Kanagy, Nancy L. ; Ansari, Maria N. ; Ghosh, Sanjay ; Webb, R. Clinton. / Recycling and buffering of intracellular calcium in vascular smooth muscle from genetically hypertensive rats. In: Journal of hypertension. 1994 ; Vol. 12, No. 12. pp. 1365-1372.
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N2 - Objective: To test the hypothesis that impaired Ca2+ recycling by the sarcoplasmic reticulum Ca-ATPase contributes to augmented force development in arteries from stroke-prone spontaneously hypertensive rats (SHRSP). Methods: Force development to caffeine (0.3-30 mmol/l) in the absence of extracellular Ca2+ was compared in aortic strips from SHRSP and Wistar-Kyoto (WKY) rats. In another protocol the strips were rinsed at the peak of contraction to caffeine (20 mmol/l) and subsequently restimulated with the alkaloid. The second response, dependent on recycled Ca2+, was used as a measure of sarcoplasmic reticulum function. A third protocol evaluated caffeine-induced contractions after Ca2+ depletion and reloading. In these latter experiments the effects of thapsigargin, an inhibitor of the sarcoplasmic reticulum Ca-ATPase, and ryanodine, an activator of sarcoplasmic reticulum Ca2+ release channels, were used to evaluate Ca2+ buffering. Finally, unidirectional 45Ca2+ influx was measured. Results: Contractions to caffeine (0.3-30 mmol/l) were larger in SHRSP aortic strips than in WKY rat strips. After a rinse at the peak of the initial response to caffeine, SHRSP segments contracted more when challenged a second time. Thapsigargin (0.3-10 mol/l) caused a concentration-dependent contraction during Ca2+ loading that was greater in SHRSP than in WKY rat strips, and a concentration-dependent inhibition of caffeine-induced contraction with similar median inhibitory concentrations in the two groups. Ryanodine did not cause contraction during Ca2+ loading, but caffeine-induced contractions were reduced after ryanodine treatment in both groups. 45Ca2+ influx was increased in SHRSP aortic segments. Conclusions: The greater force development to caffeine in SHRSP aortic strips probably reflects a greater storage of activator Ca2+ in the sarcoplasmic reticulum. On the basis of the pharmacological properties of thapsigargin and ryanodine, it appears that the larger store is caused by enhanced Ca2+ influx across the sarcolemma rather than by recycling of Ca2+ by sarcoplasmic reticulum Ca-ATPase. Experiments evaluating the secondary response to caffeine also support the interpretation that recycling of activator Ca2+ into the sarcoplasmic reticulum does not explain the augmented force development in SHRSP aortic segments.

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