Monitoring exocytosis and release from individual mast cells by capillary electrophoresis with laser-induced native fluorescence detection.
Monitoring exocytosis and release from individual mast cells by capillary electrophoresis with laser-induced native fluorescence detection.
复制标题
通过毛细管电泳和激光诱导天然荧光检测监测单个肥大细胞的胞吐作用和释放。
DOI：
10.1021/ac960412j
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发表时间：
1996
影响因子：
7.4
通讯作者：
McCloskey,MA
中科院分区：
文献类型：
--
作者：
Lillard,SJ;Yeung,ES;McCloskey,MA
使用毛细管电泳监测单个大鼠腹膜肥大细胞 (RPMC) 的柱上胞吐释放血清素和蛋白质的复杂时间演变。使用 275 nm 激发的激光诱导天然荧光检测，血清素的检测限为 1.7 amol（S/N = 3；rms）。使用生理运行缓冲液来确保细胞始终保持活力。促分泌剂是硫酸多粘菌素B (Pmx)。将单个肥大细胞注射到毛细血管中后，Pmx 向并经过细胞的电迁移诱导脱颗粒和血清素的释放。释放的时间过程以亚秒分辨率记录在电泳图中。随后引入 SDS 导致细胞完全裂解，并可以对残留的血清素进行定量。每个 RPMC 观察到的平均血清素量为 1.6 ± 0.6 fmol；血清素释放的平均百分比为 28 ± 14%。与单个亚微米颗粒（每个 250 aL）释放的血清素一致的事件是明显的，每个颗粒的平均含量为 5.9 ± 3 amol。
The complex temporal evolution of on-column exocytotic release of serotonin and proteins from individual rat peritoneal mast cells (RPMCs) was monitored by using capillary electrophoresis. Laser-induced native fluorescence detection with 275-nm excitation was used, and a detection limit of 1.7 amol (S/N = 3; rms) was obtained for serotonin. A physiological running buffer was used to ensure that the cell remained viable throughout. The secretagogue was polymyxin B sulfate (Pmx). Following the injection of a single mast cell into the capillary, electromigration of Pmx toward and past the cell induced degranulation and release of serotonin. The time course of release was registered in the electropherograms with subsecond resolution. Subsequent introduction of SDS caused the cell to lyse completely and allowed the residual serotonin to be quantified. The average amount of serotonin observed per RPMC was 1.6 ± 0.6 fmol; the average percentage of serotonin released was 28 ± 14%. Events that are consistent with released serotonin from single submicrometer granules (250 aL each) were evident, each of which contained an average amount of 5.9 ± 3 amol.