Direct demonstration of increased intracellular concentration of free calcium as measured by quin-2 in stimulated rat peritoneal mast cell.
Direct demonstration of increased intracellular concentration of free calcium as measured by quin-2 in stimulated rat peritoneal mast cell.
复制标题
通过 quin-2 在受刺激的大鼠腹膜肥大细胞中测量，直接证明细胞内游离钙浓度增加。
DOI：
10.1073/pnas.81.13.3978
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发表时间：
1984
影响因子：
11.1
通讯作者：
Sha'afi,R
中科院分区：
文献类型：
--
作者：
White,JR;Ishizaka,T;Ishizaka,K;Sha'afi,R
大鼠肥大细胞，被动致敏的单克隆小鼠IgE抗体，刺激与多价抗原，并通过使用荧光探针quin-2测定细胞质中的Ca 2+的增加。quin-2荧光的增加在抗原攻击后20秒内达到最大值，然后逐渐下降。在EGTA存在下观察到quin-2荧光显着增加，表明抗原对细胞结合的IgE抗体分子的桥接不仅诱导Ca 2+内流，还诱导细胞内Ca 2+动员。磷脂酰丝氨酸添加到培养基中增强抗原诱导的组胺释放和quin-2荧光的增加，并减慢quin-2信号返回到基础水平的速率。抗原诱导的quin-2荧光和组胺释放的增加被抑制肥大细胞与甲基转移酶，茶碱，或色甘酸抑制剂的预处理。甲基转移酶抑制剂和茶碱不仅抑制刺激依赖性钙内流，而且抑制细胞内钙库结合钙的释放。其他促分泌素，化合物48/80（1微克/毫升）和钙离子载体A23187（0.1微M），诱导大鼠肥大细胞胞质钙离子的快速增加，随后组胺释放。相反，辅致癌化合物佛波醇12-肉豆蔻酸酯13-乙酸酯引起组胺释放，而不增加quin-2荧光。
Rat mast cells, passively sensitized with monoclonal mouse IgE antibody, were stimulated with multi-valent antigen, and an increase in cytoplasmic Ca2+ was determined by using the fluorescent probe quin-2. The increase in quin-2 fluorescence reached maximum within 20 sec after the antigen challenge and then gradually declined. A substantial increase in quin-2 fluorescence was observed in the presence of EGTA, indicating that bridging of cell-bound IgE antibody molecules by antigen induced not only Ca2+ influx but also mobilization of intracellular Ca2+. Phosphatidylserine added to the medium enhanced both the antigen-induced histamine release and the increase in quin-2 fluorescence and slowed the rate at which the quin-2 signal returned to basal levels. Both the antigen-induced increase in quin-2 fluorescence and histamine release were inhibited by pretreatment of mast cells with inhibitors of methyltransferases, theophylline, or cromoglycate. It was also found that methyltransferase inhibitors and theophylline inhibited not only stimulus-dependent calcium influx but also release of bound calcium from intracellular stores. Other secretagogues, compound 48/80 (1 microgram/ml) and Ca ionophore A23187 (0.1 microM), induced a rapid increase in cytoplasmic Ca2+ in rat mast cells and subsequent histamine release. In contrast, the cocarcinogenic compound phorbol 12-myristate 13-acetate caused histamine release without increasing the quin-2 fluorescence.