The fluorescence response of chlorotetracycline-loaded human neutrophils. Correlations with lysosomal enzyme release and evidence for a 'trigger pool' of calcium.
The fluorescence response of chlorotetracycline-loaded human neutrophils. Correlations with lysosomal enzyme release and evidence for a 'trigger pool' of calcium.
复制标题
负载金四环素的人中性粒细胞的荧光响应。
DOI：
10.1016/0304-4165(82)90283-5
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发表时间：
1982
期刊：
影响因子：
--
通讯作者：
Weissmann,G
中科院分区：
文献类型：
--
作者：
Smolen,JE;Eisenstat,BA;Weissmann,G
用氯四环素（通常用作膜结合钙的探针）标记的中性粒细胞在暴露于ton -甲酰基蛋氨酸-亮基苯丙氨酸（超过1nm）时荧光迅速下降。这种下降在1分钟时达到最大，随后在3分钟时部分恢复。当中性粒细胞被n -甲酰基蛋氨酸-亮基苯丙氨酸刺激，然后在3分钟后再次暴露于相同的刺激时，观察到氯四环素荧光的进一步减少。第二次反应的强度与初始刺激的浓度成反比。同样，中性粒细胞暴露于n -甲酰基甲硫酰基亮基苯丙氨酸后，在细胞松弛素B的存在下再受n -甲酰基甲硫酰基亮基苯丙氨酸刺激，分泌氮中性粒细胞颗粒酶β-葡糖醛酸酶；酶的释放也与n -甲酰基蛋氨酸亮基苯丙氨酸的初始浓度呈负相关。这些反应也有时间依赖性。氯四环素荧光和β-葡萄糖醛酸酶释放的二次衰减随两次n -甲酰基蛋氨酸-亮基苯丙氨酸给药时间的延长而增加。相比之下，肉豆蔻酸酯佛波引起的氯四环素荧光下降没有类似的恢复期。当中性粒细胞受到肉豆酸酯佛波刺激后，暴露于ton -甲酰基甲硫酰基亮基苯丙氨酸时，随着两次刺激间隔时间的增加，氯四环素荧光的第二次衰减减弱。随着初始刺激时间的延长，β-葡萄糖醛酸酶的分泌也随之减少。当n -甲酰基蛋氨酸-亮基苯丙氨酸作为初始刺激时，肉豆蔻酸酯佛波的氯四环素荧光响应特性至少在1min内无法观察到。这些结果与假设是一致的，即氯四环素作为可移动的膜结合“触发钙”的探针，一个充满的池是溶酶体酶释放的必要条件。
Neutrophils labelled with chlorotetracycline (commonly employed as a probe for membrane-bound calcium), underwent rapid decreases in fluorescence upon exposure toN-formylmethionylleucylphenylalanine (morethan1nM). This decrease was maximal at 1 min and was followed by partial recovery by 3 min. When neutrophils were stimulated withN-formylmethionylleucylphenylalanine and then re-exposed to the same stimulus 3 min later, an additional decrease in chlorotetracycline fluorescence was observed. The magnitude of this second response was inversely related to the concentration of the initial stimulus. Similarly, neutrophils exposed toN-formylmethionylleucylphenylalanine and then restimulated byN-formylmethionylleucylphenylalanine in the presence of cytochalasin B secreted the azurophil granule enzymeβ-glucuronidase; release of the enzyme was also inversely related to the initial concentration ofN-formylmethionylleucylphenylalanine. These responses were also time-dependent. Both the second decrement in chlorotetracycline fluorescence andβ-glucuronidase release increased with time allowed between the two administrations ofN-formylmethionylleucylphenylalanine. In contrast, decreases in chlorotetracycline fluorescence induced by phorbol myristate acetate showed no comparable recovery phase. When neutrophils, stimulated with phorbol myristate acetate, were then exposed toN-formylmethionylleucylphenylalanine, the second decrement in chlorotetracycline fluorescence diminished as the time allowed between the two stimuli was increased. Secretion ofβ-glucuronidase in response toN-formylmethionylleucylphenylalanine was also diminished by increasing the time of exposure to the initial stimulus of phorbol myristate acetate. WhenN-formylmethionylleucylphenylalanine was used as the initial stimulus, the chlorotetracycline fluorescence response characteristic of phorbol myristate acetate could not be observed for at least 1 min. These results are consistent with the hypothesis that chlorotetracycline serves as a probe of mobilizable membrane-bound ‘trigger calcium’, a replete pool of which is an obligate requirement for lysosomal enzyme release.