TIRF imaging of docking and fusion of single insulin granule motion in primary rat pancreatic β-cells:: different behaviour of granule motion between normal and Goto-Kakizaki diabetic rat β-cells
TIRF imaging of docking and fusion of single insulin granule motion in primary rat pancreatic β-cells:: different behaviour of granule motion between normal and Goto-Kakizaki diabetic rat β-cells
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DOI：
10.1042/bj20040434
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发表时间：
2004-07-01
影响因子：
4.1
通讯作者：
Nagamatsu, S
中科院分区：
文献类型：
--
作者：
Mica, OI;Nishiwaki, C;Nagamatsu, S
我们使用全内反射荧光显微镜（TIRFM）成像并分析了正常和糖尿病Goto-Kakizaki （GK）大鼠活胰腺β细胞中靠近质膜的单个胰岛素分泌颗粒的运动。在正常大鼠原代β细胞中，在第一阶段融合的颗粒来自先前停靠的颗粒，而在第二阶段的颗粒来自“新来者”。在糖尿病GK大鼠β细胞中，先前停靠颗粒的融合事件数量明显减少，相反，新颗粒的融合仍然保留。葡萄糖刺激后，GK大鼠β细胞中胰岛素停靠颗粒数量的动态变化表明，胰岛素停靠颗粒总数明显减少至初始数量的35%。TIRFM观察抗胰岛素抗体免疫组化显示，GK大鼠β细胞停靠在质膜上的内源性胰岛素颗粒明显减少。因此，我们的研究结果表明，在糖尿病GK大鼠β细胞胰岛素释放的第一阶段，停靠胰岛素颗粒数量的减少是胰岛素释放受损的原因。
We imaged and analysed the motion of single insulin secretory granules near the plasma membrane in live pancreatic beta-cells, from normal and diabetic Goto-Kakizaki (GK) rats, using total internal reflection fluorescence microscopy (TIRFM). In normal rat primary beta-cells, the granules that were fusing during the first phase originate from previously docked granules, and those during the second phase originate from 'newcomers'. In diabetic GK rat beta-cells, the number of fusion events from previously docked granules were markedly reduced, and, in contrast, the fusion from newcomers was still preserved. The dynamic change in the number of docked insulin granules showed that, in GK rat beta-cells, the total number of docked insulin granules was markedly decreased to 35 % of the initial number after glucose stimulation. Immunohistochemistry with anti-insulin antibody observed by TIRFM showed that GK rat beta-cells had a marked decline of endogenous insulin granules docked to the plasma membrane. Thus our results indicate that the decreased number of docked insulin granules accounts for the impaired insulin release during the first phase of insulin release in diabetic GK rat beta-cells.