Targeting of green fluorescent protein to secretory granules in oxytocin magnocellular neurons and its secretion from neurohypophysial nerve terminals in transgenic mice.
Targeting of green fluorescent protein to secretory granules in oxytocin magnocellular neurons and its secretion from neurohypophysial nerve terminals in transgenic mice.
复制标题
DOI：
10.1210/endo.143.3.8700
复制
发表时间：
2002-03
期刊：
影响因子：
4.8
通讯作者：
B-J. Zhang;K. Kusano;P. Zerfas;A. Iacangelo;W. Young;H. Gainer
中科院分区：
文献类型：
--
作者：
B-J. Zhang;K. Kusano;P. Zerfas;A. Iacangelo;W. Young;H. Gainer
催产素（OT）是一种下丘脑九肽，作为一种更大的前体蛋白的一部分合成，该前体蛋白在其C-末端还含有一种约10 kDa的称为neurophysin的蛋白质。这种前体蛋白通过受调节的分泌途径进入分泌颗粒，然后经轴突运输到垂体神经叶中的神经末梢并从其分泌。在本文中，我们表明，AI-03转基因，其中包含增强的绿色荧光蛋白（EGFP）融合的神经垂体素的C-末端的OT前激素原，选择性地表达在OT-大细胞神经元，并被贩运到转基因小鼠的分泌颗粒。然后将含有EGFP的分泌颗粒运输到神经垂体中的OT-神经分泌末端，在那里EGFP荧光经历去极化诱导的钙依赖性分泌。在神经叶的内源性荧光是足够强的图像分泌事件在个别OT神经末梢（神经分泌体）分离的后垂体在这些转基因小鼠。
Oxytocin (OT) is a hypothalamic nonapeptide that is synthesized as part of a larger precursor protein that also contains an approximately 10-kDa protein called neurophysin at its C-terminus. This precursor protein is trafficked through the regulated secretory pathway into secretory granules and then axonally transported to and secreted from nerve terminals in the neural lobe of the pituitary. In this paper, we show that the AI-03 transgene that contains enhanced green fluorescent protein (EGFP) fused to the end of the neurophysin at the C-terminus of the OT pre-prohormone, is expressed selectively in OT-magnocellular neurons and is trafficked to secretory granules in transgenic mice. The EGFP-containing secretory granules are then transported to OT-neurosecretory terminals in the neurohypophysis, where the EGFP fluorescence undergoes depolarization-induced calcium-dependent secretion. The endogenous fluorescence in the neural lobes is sufficiently intense to image secretory events in individual OT nerve terminals (neurosecretosomes) isolated from the posterior pituitaries in these transgenic mice.