Cell culture alters Ca2+ entry pathways activated by store-depletion or hypoxia in canine pulmonary arterial smooth muscle cells.

Cell culture alters Ca2+ entry pathways activated by store-depletion or hypoxia in canine pulmonary arterial smooth muscle cells. Research Abstract Details 

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Cell culture alters Ca2+ entry pathways activated by store-depletion or hypoxia in canine pulmonary arterial smooth muscle cells. Abstract Text:

Lih Chyuan Ng,Barry D Kyle,Alison R Lennox,Xiao-Ming Shen,William J Hatton,Joseph R Hume,Lih Chyuan Ng,Barry D Kyle,Alison R Lennox,Xiao-Ming Shen,William J Hatton,Joseph R Hume,Lih Chyuan Ng,Barry D Kyle,Alison R Lennox,Xiao-Ming Shen,William J Hatton,Joseph R Hume,

Previous studies have shown that, in acutely dispersed canine pulmonary artery smooth muscle cells (PASMCs), depletion of both functionally independent inositol 1,4,5-trisphosphate (IP(3))- and ryanodine-sensitive Ca(2+) stores activates capacitative Ca(2+) entry (CCE). The present study aimed to determine if cell culture modifies intracellular Ca(2+) stores and alters Ca(2+) entry pathways caused by store depletion and hypoxia in canine PASMCs. Intracellular Ca(2+) concentration ([Ca(2+)](i)) was measured in fura 2-loaded cells. Mn(2+) quench of fura 2 signal was performed to study divalent cation entry, and the effects of hypoxia were examined under oxygen tension of 15-18 mmHg. In acutely isolated PASMCs, depletion of IP(3)-sensitive Ca(2+) stores with cyclopiazonic acid (CPA) did not affect initial caffeine-induced intracellular Ca(2+) transients but abolished 5-HT-induced Ca(2+) transients. In contrast, CPA significantly reduced caffeine- and 5-HT-induced Ca(2+) transients in cultured PASMCs. In cultured PASMCs, store depletion or hypoxia caused a transient followed by a sustained rise in [Ca(2+)](i). The transient rise in [Ca(2+)](i) was partially inhibited by nifedipine, whereas the nifedipine-insensitive transient rise in [Ca(2+)](i) was inhibited by KB-R7943, a selective inhibitor of reverse mode Na(+)/Ca(2+) exchanger (NCX). The nifedipine-insensitive sustained rise in [Ca(2+)](i) was inhibited by SKF-96365, Ni(2+), La(3+), and Gd(3+). In addition, store depletion or hypoxia increased the rate of Mn(2+) quench of fura 2 fluorescence that was also inhibited by these blockers, exhibiting pharmacological properties characteristic of CCE. We conclude that cell culture of canine PASMCs reorganizes IP(3) and ryanodine receptors into a common intracellular Ca(2+) compartment, and depletion of this store or hypoxia activates voltage-operated Ca(2+) entry, reverse mode NCX, and CCE.

Cell culture alters Ca2+ entry pathways activated by store-depletion or hypoxia in canine pulmonary arterial smooth muscle cells. Publishing Authors By Initials

LC Ng,BD Kyle,AR Lennox,XM Shen,WJ Hatton,JR Hume,LC Ng,BD Kyle,AR Lennox,XM Shen,WJ Hatton,JR Hume,LC Ng,BD Kyle,AR Lennox,XM Shen,WJ Hatton,JR Hume,

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Cell culture alters Ca2+ entry pathways activated by store-depletion or hypoxia in canine pulmonary arterial smooth muscle cells. Journal Published:

PUBLICATION TYPE: Journal Article

Journal: American journal of physiology. Cell physiology

VOLUME: 294

Page Numbers: C313-23

Journal Abbreviation: Am. J. Physiol., Cell Physiol.

ISSN: 0363-6143

DAY: 31

MONTH: 10

YEAR: 2007

Cell culture alters Ca2+ entry pathways activated by store-depletion or hypoxia in canine pulmonary arterial smooth muscle cells. Information

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LANGUAGE: eng

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AFFILIATION: Dept. of Pharmacology/318, Univ. of Nevada School of Medicine, 1664 North Virginia St., Reno, NV 89557. joeh@med.unr.edu).

Country: United States

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MEDLINETA: Am J Physiol Cell Physiol

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