Differential exocytosis from human endothelial cells evoked by high intracellular Ca2+ concentration

被引:30
作者
Zupancic, G
Ogden, D
Magnus, CJ
Wheeler-Jones, C
Carter, TD
机构
[1] UCL, Dept Pharmacol, London WC1E 6BT, England
[2] Natl Inst Med Res, London NW7 1AA, England
[3] Univ London Royal Vet Coll, Dept Vet Basic Sci, London NW1 0TU, England
[4] Univ Ljubljana, Dept Biol, Ljubljana 1001, Slovenia
来源
JOURNAL OF PHYSIOLOGY-LONDON | 2002年 / 544卷 / 03期
关键词
D O I
10.1113/jphysiol.2002.027490
中图分类号
Q189 [神经科学];
学科分类号
071006 ;
摘要
Endothelial cells secrete a range of procoagulant, anticoagulant and inflammatory proteins by exocytosis to regulate blood clotting and local immune responses. The mechanisms regulating vesicular exocytosis were studied in human umbilical vein endothelial cells (HUVEC) with high-resolution membrane capacitance (C(m)) measurements. The total whole-cell C(m) and the amplitudes and times of discrete femtoFarad (fF)-sized C(m) steps due to exocytosis and endocytosis were monitored simultaneously. Intracellular calcium concentration [Ca(2+)](i) was elevated by intracellular photolysis of calcium-DM-nitrophen to evoke secretion and monitored with the low-affinity Ca(2+) indicator faraptra. Sustained elevation of [Ca(2+)](i) to > 20 pm evoked large, slow increases in C(m) of up to 5 pF in 1-2 min. Exocytotic and endocytotic steps of amplitude 0.5-110 fF were resolved, and accounted on average for similar to33% of the total C(m) change. A prominent component of C(m) steps of 2.5-9.0 fF was seen and could be attributed to exocytosis of von-Willebrand-factor-containing Weibel-Palade bodies (WPb), based on the near-identical distributions of capacitance step amplitudes, with calculated estimates of WPb capacitance from morphometry, and on the absence of 2.5-9.0 fF C(m) steps in cells deficient in WPb. WPb secretion was delayed on average by 23 s after [Ca(2+)](i) elevation, whereas total C(m) increased immediately due to the secretion of small, non-WPb granules. The results show that following a large increase of [Ca(2+)](i), corresponding to strong stimulation, small vesicular components are immediately available for secretion, whereas the large WPb undergo exocytosis only after a delay. The presence of events of magnitude 9-110 fF also provides evidence of compound secretion of WPb due to prior fusion of individual granules.
引用
收藏
页码:741 / 755
页数:15
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