Imaging exocytosis of single insulin secretory granules with evanescent wave microscopy - Distinct behavior of granule motion in biphasic insulin release
Imaging exocytosis of single insulin secretory granules with evanescent wave microscopy - Distinct behavior of granule motion in biphasic insulin release
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DOI：
10.1074/jbc.c100712200
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发表时间：
2002-02-08
影响因子：
4.8
通讯作者：
Nagamatsu, S
中科院分区：
文献类型：
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作者：
Ohara-Imaizumi, M;Nakamichi, Y;Nagamatsu, S
为了通过监测单个胰岛素分泌颗粒运动来研究胰岛素胞吐作用，使用evaneScent波显微镜用双相释放来定量分析胰岛素胞吐作用的最后阶段。在MIN6 Beta细胞中转染的绿色荧光蛋白标记的胰岛素被包装在胰岛素分泌颗粒中，似乎优先将其停靠在质膜上。促进g的融合后，淡波显微镜表明，绿色荧光蛋白标记的胰岛素的荧光变亮，扩散（300毫秒内），然后消失。在代表释放第一阶段的KCL刺激下，连续的融合事件主要是从先前停靠的颗粒中看到的。然后将新颗粒招募到血浆对接地点。相比之下，葡萄糖用葡萄糖刺激在前120秒内引起了先前停靠颗粒的融合事件，此后观察到连续融合（第二阶段释放）在10分钟内观察到了10分钟，主要是从新募集的颗粒逐渐积累的颗粒。因此，我们的数据揭示了在释放的第一阶段和第二阶段中胰岛素颗粒运动的独特行为。
To study insulin exocytosis by monitoring the single insulin secretory granule motion, evanescent wave microscopy was used to quantitatively analyze the final stage of insulin exocytosis with biphasic release. Green fluorescent protein-tagged insulin transfected in MIN6 beta cells was packed in insulin secretory granules, which appeared to preferentially dock to the plasma membrane. Upon fusion evoked by secretagogues, evanescent wave microscopy revealed that fluorescence of green fluorescent protein-tagged insulin brightened, spread (within 300 ms), and then vanished. Under KCl stimulation, which represents the 1st phase of release, the successive fusion events were seen mostly from previously docked granules for the first minute; followed by the recruitment of new granules to the plasmalemmal docking sites. Stimulation with glucose, in contrast, caused the fusion events from previously docked granules for the first 120 s, thereafter a continuous fusion (2nd phase of release) was observed over 10 min mostly from newly recruited granules that progressively accumulated on the plasma membrane. Thus, our data revealed the distinct behavior of the insulin granule motion during the 1st and 2nd phase of release.