Functional molecular morphology of anterior pituitary cells, from hormone production to intracellular transport and secretion
Functional molecular morphology of anterior pituitary cells, from hormone production to intracellular transport and secretion
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垂体前叶细胞的功能分子形态，从激素产生到细胞内运输和分泌
DOI：
10.1007/s00795-011-0545-4
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发表时间：
2011
期刊：
影响因子：
1.8
通讯作者：
R.Y.
中科院分区：
文献类型：
--
作者：
Matsuno;A.;Mizutani;A.;Okinaga;H.;Takano;K.;Yamada;S.;Yamada;S.M.;Nakaguchi;H.;Hoya;K.;Murakami;M.;Takeuchi;M.;Sugaya;M.;Itoh;J.;Takekoshi;S.;Osamura;R.Y.
电子显微镜下的原位杂交 (ISH) 和免疫组织化学 (IHC) 组合 (EM-ISH & IHC) 具有足够的超微结构分辨率，可以提供垂体细胞中垂体激素及其 mRNA 亚细胞定位的二维图像。半导体纳米晶体（量子点；Qdots）和共焦激光扫描显微镜（CLSM）的优势使我们能够获得垂体激素及其mRNA亚细胞定位的三维图像。 EM-ISH & IHC 以及使用 Qdots 和 CLSM 的 ISH & IHC 都有助于在二维或三维上同时了解蛋白质和 mRNA 之间的关系。对 rab3B 和 SNARE 蛋白（例如 SNAP-25 和 Syntaxin）的 CLSM 观察表明，rab3B 和 SNARE 系统蛋白都发挥着重要作用，并作为垂体前叶细胞中的胞吐机制共同发挥作用。另一个重要问题是垂体激素的细胞内运输和分泌。已经开发出一种实验性垂体细胞系 GH3 细胞，其中生长激素 (GH) 与增强型黄色荧光素蛋白 (EYFP) 连接。这种稳定的 GH3 细胞在受到 Ca2+ 流入或储存的 Ca2+ 释放的刺激后，会分泌与 EYFP 连接的 GH。该 GH3 细胞可用于 GH 细胞内运输和分泌的实时可视化。这三种方法使我们能够连续地可视化垂体激素的转录、翻译、运输和分泌过程。
Combined in situ hybridization (ISH) and immunohistochemistry (IHC) under electron microscopy (EM-ISH & IHC) has sufficient ultrastructural resolution to provide two-dimensional images of subcellular localization of pituitary hormone and its mRNA in a pituitary cell. The advantages of semiconductor nanocrystals (Quantum dots; Qdots) and confocal laser scanning microscopy (CLSM) enable us to obtain three-dimensional images of the subcellular localization of pituitary hormone and its mRNA. Both EM-ISH & IHC and ISH & IHC using Qdots and CLSM are useful for understanding the relationship between protein and mRNA simultaneously in two or three dimensions. CLSM observation of rab3B and SNARE proteins such as SNAP-25 and syntaxin revealed that both rab3B and SNARE system proteins play an important role and work together as the exocytotic machinery in anterior pituitary cells. Another important issue is the intracellular transport and secretion of pituitary hormone. An experimental pituitary cell line, the GH3 cell, in which growth hormone (GH) is linked to enhanced yellow fluorescein protein (EYFP), has been developed. This stable GH3 cell secretes GH linked to EYFP upon being stimulated by Ca2+influx or Ca2+release from storage. This GH3 cell is useful for real-time visualization of the intracellular transport and secretion of GH. These three methods enable us to visualize consecutively the processes of transcription, translation, transport, and secretion of pituitary hormone.