TY - JOUR
T1 - The sources and sequestration of Ca2+ contributing to neuroeffector Ca2+ transients in the mouse vas deferens
AU - Brain, Keith
AU - Cuprian, AM
AU - Williams, DJ
AU - Cunnane, TC
PY - 2003/12/1
Y1 - 2003/12/1
N2 - The detection of focal Ca2+ transients (called neuroeffector Ca2+ transients, or NCTs) in smooth muscle of the mouse isolated vas deferens has been used to detect the packeted release of ATP from nerve terminal varicosities acting at postjunctional P2X receptors. The present study investigates the sources and sequestration of Ca2+ in NCTs. Smooth muscle cells in whole mouse deferens were loaded with the Ca2+ indicator Oregon Green 488 BAPTA-1 AM and viewed with a confocal microscope. Ryanodine (10 muM) decreased the amplitude of NCTs by 45 +/- 6%. Cyclopiazonic acid slowed the recovery of NCTs (from a time course of 200 +/- 10 ms to 800 +/- 100 ms). Caffeine (3 mm) induced spontaneous focal smooth muscle Ca2+ transients (sparks). Neither of the T-type Ca2+ channel blockers NiCl2 (50 muM) or mibefradil dihydrochloride (10 muM) affected the amplitude of excitatory junction potentials (2 +/- 5% and -3 +/- 10%) or NCTs (-20 +/- 36% and 3 +/- 13%). In about 20% of cells, NCTs were associated with a local, subcellular twitch that remained in the presence of the alpha(1)-adrenoceptor antagonist prazosin (100 nM), showing that NCTs can initiate local contractions. Slow (5.8 +/- 0.4 mum s(-1)), spontaneous smooth muscle Ca2+ waves were occasionally observed. Thus, Ca2+ stores initially amplify and then sequester the Ca2+ that enters through P2X receptors and there is no amplification by local voltage-gated Ca2+ channels.
AB - The detection of focal Ca2+ transients (called neuroeffector Ca2+ transients, or NCTs) in smooth muscle of the mouse isolated vas deferens has been used to detect the packeted release of ATP from nerve terminal varicosities acting at postjunctional P2X receptors. The present study investigates the sources and sequestration of Ca2+ in NCTs. Smooth muscle cells in whole mouse deferens were loaded with the Ca2+ indicator Oregon Green 488 BAPTA-1 AM and viewed with a confocal microscope. Ryanodine (10 muM) decreased the amplitude of NCTs by 45 +/- 6%. Cyclopiazonic acid slowed the recovery of NCTs (from a time course of 200 +/- 10 ms to 800 +/- 100 ms). Caffeine (3 mm) induced spontaneous focal smooth muscle Ca2+ transients (sparks). Neither of the T-type Ca2+ channel blockers NiCl2 (50 muM) or mibefradil dihydrochloride (10 muM) affected the amplitude of excitatory junction potentials (2 +/- 5% and -3 +/- 10%) or NCTs (-20 +/- 36% and 3 +/- 13%). In about 20% of cells, NCTs were associated with a local, subcellular twitch that remained in the presence of the alpha(1)-adrenoceptor antagonist prazosin (100 nM), showing that NCTs can initiate local contractions. Slow (5.8 +/- 0.4 mum s(-1)), spontaneous smooth muscle Ca2+ waves were occasionally observed. Thus, Ca2+ stores initially amplify and then sequester the Ca2+ that enters through P2X receptors and there is no amplification by local voltage-gated Ca2+ channels.
U2 - 10.1113/jphysiol.2003.049734
DO - 10.1113/jphysiol.2003.049734
M3 - Article
C2 - 14500773
SN - 1469-7793
VL - 553
SP - 627
EP - 635
JO - The Journal of Physiology
JF - The Journal of Physiology
IS - 2
ER -