Imaging of Zn2+ release from pancreatic beta-cells at the level of single exocytotic events.
Imaging of Zn2+ release from pancreatic beta-cells at the level of single exocytotic events.
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在单个胞吐事件水平上从胰腺 β 细胞释放 Zn2 的成像。
DOI：
10.1021/ac0341057
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发表时间：
2003
影响因子：
7.4
通讯作者：
Kennedy,RobertT
中科院分区：
文献类型：
--
作者：
Qian,Wei-Jun;Gee,KyleR;Kennedy,RobertT
使用激光扫描共聚焦显微镜对来自分离的胰腺β细胞的Zn 2+的调节分泌进行成像。在该方法中，将β细胞在含有新型荧光Zn 2+指示剂FluoZin-3的溶液中孵育。共聚焦显微镜下观察到细胞释放的Zn 2+与染料反应生成荧光产物。新染料比以前用于该应用的Zinquin更亮，检测限为10 - 40 nM，时间分辨率为16 ms/图像。高时间分辨率允许在用20 mM葡萄糖或40 mM K+刺激后在细胞边缘发生的孤立荧光瞬变的成像。荧光瞬变从初始上升到峰值需要16 - 50 ms，并在170 ± 50 ms后返回基线（n= 78个瞬变，来自15个细胞）。得出的结论是，瞬态对应于检测Zn 2+的胞吐释放。的瞬态的时间和空间分散的分析表明，锌2+的释放是不扩散限制，而是动力学控制与以前的观察电流法检测到的胰岛素释放。
Regulated secretion of Zn2+from isolated pancreatic β-cells was imaged using laser-scanning confocal microscopy. In the method, β-cells were incubated in a solution containing the novel fluorescent Zn2+indicator FluoZin-3. Zn2+released from the cells reacted with the dye to form a fluorescent product, which was detected by the confocal microscope. The new dye is much brighter than Zinquin, previously used for this application, allowing detection limits of 10−40 nM and temporal resolution of 16 ms/image. The high temporal resolution allowed imaging of isolated fluorescent transients that occurred at the edge of the cells following stimulation with 20 mM glucose or 40 mM K+. Fluorescent transients took 16−50 ms to reach a peak from the initial rise and returned to baseline after 170 ± 50 ms (n= 78 transients from 15 cells). It was concluded that the transients correspond to detection of exocytotic release of Zn2+. Analysis of the temporal and spatial dispersion of the transients indicates that the release of Zn2+is not diffusion limited but is instead kinetically controlled in agreement with previous observations of insulin release detected by amperometry.