Preferential release of newly synthesized insulin assessed by a multi-label reporter system using pancreatic β-cell line MIN6.

Preferential release of newly synthesized insulin assessed by a multi-label reporter system using pancreatic β-cell line MIN6.
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DOI:
10.1371/journal.pone.0047921
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发表时间:
2012
期刊:
影响因子:
3.7
通讯作者:
Torii S
Torii S
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Hou N;Mogami H;Kubota-Murata C;Sun M;Takeuchi T;Torii S

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新合成的激素被认为优先由各种神经内分泌细胞分泌。这一观察结果表明,有一个独特的人口分泌颗粒含有新的和旧的激素。牛肾上腺嗜铬细胞中使用的荧光计时器蛋白的最新发展揭示了分泌囊泡分离成不同的年龄依赖性群体。在这里,我们验证了优先释放的新合成的胰岛素在胰腺β细胞系,MIN6,使用多标记报告系统与荧光和生化程序的组合。该系统允许标记任何年龄的激素或颗粒,与计时器蛋白相反,计时器蛋白需要荧光移位时间。脉冲追踪标记不同颜色的探针区分胰岛素分泌颗粒的年龄,年轻的颗粒具有主要的细胞内定位,而不是在细胞周边。
Newly synthesized hormones have been suggested to be preferentially secreted by various neuroendocrine cells. This observation indicates that there is a distinct population of secretory granules containing new and old hormones. Recent development of fluorescent timer proteins used in bovine adrenal chromaffin cells revealed that secretory vesicles segregate into distinct age-dependent populations. Here, we verify the preferential release of newly synthesized insulin in the pancreatic β-cell line, MIN6, using a combination of multi-labeling reporter systems with both fluorescent and biochemical procedures. This system allows hormones or granules of any age to be labeled, in contrast to the timer proteins, which require fluorescence shift time. Pulse-chase labeling with different color probes distinguishes insulin secretory granules by age, with younger granules having a predominantly intracellular localization rather than at the cell periphery.
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