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首页> 外文期刊>Life sciences >Subcellular distribution of choline acetyltransferase by immunogold electron microscopy in non-neuronal cells: placenta, airways and murine embryonic stem cells.
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Subcellular distribution of choline acetyltransferase by immunogold electron microscopy in non-neuronal cells: placenta, airways and murine embryonic stem cells.

机译：非神经元细胞中免疫偶极电子显微镜的胆碱乙酰转移酶的亚细胞分布：胎盘，气道和鼠胚胎干细胞。

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摘要

Acetylcholine is synthesized in more or less all mammalian cells. However, little is known about the subcellular location of acetylcholine synthesis. Therefore, in the present experiments the subcellular location of the synthesizing enzyme choline acetyltransferase (ChAT) was investigated by anti-ChAT immunogold electron microscopy in human placenta and airways as well as in a murine embryonic stem cell line (CGR8 cell line).Human tissue was obtained as so-called surplus tissue (after delivery/surgical removal because of lung tumor); the CGR8 stem cell line was cultured under standard conditions. For human tissue a monoclonal mouse anti-ChAT antibody (ab) was used and for the CGR8 cell line a polyclonal goat anti-ChAT ab. Immunogold electron microscopy was applied to identify the subcellular location of ChAT.In trophoblast cells (placenta) specific anti-ChAT immunogold deposition was found within the cell membrane, microvilli, and caveolae but also within the cytosol, for example associated with intermediate filaments. In addition, immunogold deposition was identified within mitochondria and the nuclear membrane. In airway epithelial cells anti-ChAT immunogold was found particularly within the apical cell membrane, cilia, submucosa, cytosol and nuclear membrane. Likewise alveolar macrophages showed positive anti-ChAT immunogold within the nucleus, nuclear membrane and granula. Also in the CGR8 cell line positive anti-ChAT immunogold was identified within the cell nucleus and cytosol.The present experiments demonstrate a wide subcellular distribution of ChAT with particular preference of the cell membrane in human epithelial cells.
机译：乙酰胆碱在或多或少地合成所有哺乳动物细胞。然而，关于乙酰胆碱合成的亚细胞位置很少。因此，在本实验中，通过人胎盘和呼吸道以及鼠胚胎干细胞系（CGR8细胞系），通过抗聊天免疫电解电子显微镜研究了合成酶胆碱乙酰转移酶（聊天）的亚细胞位置。获得如所谓的剩余组织（由于肺肿瘤而递送/手术移除）; CGR8干细胞系在标准条件下培养。对于人体组织，使用单克隆小鼠抗聊天抗体（AB）和用于CGR8细胞系的多克隆山羊防聊AB。施用免疫角膜电子显微镜检查亚细胞的亚细胞位置。在细胞膜，微绒毛和Caveolae中发现滋养细胞（胎盘）特异性抗聊天免疫沉积，但也发现了胞质细胞溶胶内，例如与中间细丝相关。此外，在线粒体和核膜内鉴定免疫沉积。在气道上皮细胞中，特别是在顶端细胞膜，纤毛，粘膜，细胞溶溶胶和核膜内发现抗聊天的免疫元。同样肺泡巨噬细胞在核，核膜和颗粒中显示出正抗聊天免疫形状。同样在CGR8细胞系中，在细胞核和细胞溶胶中鉴定了阳性抗聊天免疫角质。本实验表明了人上皮细胞中细胞膜的特定优选聊天的宽亚细胞分布。

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