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Am J Physiol Cell Physiol (March 5, 2003). doi:10.1152/ajpcell.00374.2002
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Submitted on August 19, 2002
Accepted on February 27, 2003

Inositol 1,4,5-trisphosphate receptors modulate Ca2+-sparks and Ca2+-store content in vas deferens myocytes

Carl White1 and J. Graham McGeown1*

1 Physiology, The Queen's University of Belast, Belfast, N. Ireland, United Kingdom

* To whom correspondence should be addressed. E-mail: g.mcgeown{at}qub.ac.uk.

Spontaneous Ca2+ sparks were observed in fluo-4 loaded myocytes from guinea-pig vas deferens using linescan confocal imaging. They were abolished by ryanodine (100 µM) but the inositol 1,4,5-trisphosphate receptor (IP3R) blockers 2-aminoethoxydiphenyl borate (2APB, 100 µM) and intracellular heparin (5 mgml-1) increased spark frequency, rise-time, duration and spread. Very prolonged Ca2+ release events were also observed in approximately 20% of cells treated with IP3R blockers but not under control conditions. 2APB and heparin abolished noradrenaline (10 µM; zero Ca2+) evoked Ca2+ transients but increased caffeine (10 mM; zero Ca2+) transients in fura-2 loaded myocytes. Transients evoked by ionomycin (25 µM; zero Ca2+) were also enhanced by 2APB. Ca2+ sparks and transients evoked by noradrenaline and caffeine were abolished by thimerosal (100 µM), which sensitizes the IP3R to IP3. In cells voltage clamped at -40 mV, spontaneous transient outward currents (STOCs) were increased in frequency, amplitude and duration in the presence of 2APB. These data are consistent with a model in which the Ca2+-store content in smooth muscle is limited by tonic release of Ca2+ via an IP3-dependent pathway. Blockade of IP3Rs elevates SR store content, promoting Ca2+ sparks and STOC activity.




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