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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Expression of γ-aminobutyric acid ρ1 and ρ1△450 as gene fusions with the green fluorescent protein
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Expression of γ-aminobutyric acid ρ1 and ρ1△450 as gene fusions with the green fluorescent protein

机译：γ-氨基丁酸ρ1和ρ1△450与绿色荧光蛋白的基因融合表达

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摘要

The functional characteristics and cellular localization of the V aminobutyric acid (GABA) ρ1 receptor and its nonfunctional isoform ρ1△450 were investigated by expressing them as gene fusions with the enhanced version of the green fluorescent protein (GFP). Oocytes injected with ρ1-GFP had receptors that gated chloride channels when activated by GABA. The functional characteristics of these receptors were the same as for those of wild-type ρ1 receptors. Fluorescence. because of the chimeric receptors expressed. was over the whole oocyte but was more intense near the cell surface and more abundant in the animal hemisphere. Similar to the wild type, ρ1 △450-GFP did not lead to the expression of functional GABA receptors, and injected oo- cytes failed to generate currents even after exposure to high concentrations of GABA. Nonetheless, the fluorescence dis- played by oocytes expressing ρ1△450-GFP was distributed sim- ilarly to that of ρ1-GFP. Mammalian cells transfected with the ρ1-GFP or ρ1△450-GFP constructs showed mostly intracellularly distributed fluorescence in confocal microscope images. A sparse localization of fluorescence was observed in the plasma mem- brane regardless of the cell line used. We conclude that ρ1△450 is expressed and transported close to, and perhaps incorporated into, the plasma membrane. Thus, ρ1- and ρ1△450-GFP fusions provide a powerful tool to visualize the traffic of GABA type C receptors.
机译：通过将它们表达为与绿色荧光蛋白（GFP）增强版本的基因融合体，研究了V氨基丁酸（GABA）ρ1受体及其非功能性同工型ρ1△450的功能特性和细胞定位。注射了ρ1-GFP的卵母细胞具有被GABA激活时可控制氯离子通道的受体。这些受体的功能特性与野生型ρ1受体相同。荧光。因为嵌合受体的表达。在整个卵母细胞中，但在细胞表面附近更强烈，在动物半球中更丰富。与野生型相似，ρ1△450-GFP不会导致功能性GABA受体的表达，即使在暴露于高浓度的GABA后，注入的卵细胞也无法产生电流。尽管如此，表达ρ1△450-GFP的卵母细胞所显示的荧光与ρ1-GFP相似。用ρ1-GFP或ρ1△450-GFP构建体转染的哺乳动物细胞在共聚焦显微镜图像中显示出大部分细胞内分布的荧光。无论使用哪种细胞系，在血浆膜中均观察到荧光的稀疏定位。我们得出的结论是，ρ1△450在质膜附近表达并运输，也许掺入到质膜中。因此，ρ1-和ρ1△450-GFP融合蛋白为可视化GABA C型受体的运输提供了强大的工具。

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