Liposome-mediated transfection of mature taste cells.
Liposome-mediated transfection of mature taste cells.
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脂质体介导的成熟味觉细胞转染。
DOI：
10.1002/neu.20157
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发表时间：
2005
期刊：
影响因子：
--
通讯作者：
Chaudhari,Nirupa
中科院分区：
文献类型：
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作者：
Landin,AnaMarie;Kim,JoungWoul;Chaudhari,Nirupa
外源DNA在神经元中的引入和表达对于分析外周的一系列细胞和分子过程很有价值，例如转导相关蛋白的作用、生长因子对发育和分化的影响以及特定于细胞类型的启动子的功能。然而，感觉受体细胞，特别是化学感应细胞，很难转染。我们已成功将表达绿色和 Discosoma 红色荧光蛋白（GFP 和 DsRed）的质粒引入原代培养的大鼠味蕾中。当用新鲜蛋白酶重新消化分层的味觉上皮时，转染效率增加，这表明味觉细胞周围的细胞外基质的保护屏障通常可能存在。由于味蕾是细胞的异质聚集体，因此我们使用α-味觉素、神经元细胞粘附分子 (NCAM) 和神经元泛素羧基末端水解酶 (PGP9.5)（成熟味觉细胞特定亚群的标记物）来证明脂质体介导的转染针对多种味觉细胞类型。在测试了八种市售脂质后，我们确定了一种对味觉细胞最有效的 Transfast。我们还证明了两种常见的“混杂”启动子和一种味觉细胞内源使用的启动子的有效性。这些研究应该允许研究味觉细胞发育和细胞功能的离体策略。 © 2005 Wiley periodicals, Inc. J. Neurobiol，2005
The introduction and expression of exogenous DNA in neurons is valuable for analyzing a range of cellular and molecular processes in the periphery, e.g., the roles of transduction‐related proteins, the impact of growth factors on development and differentiation, and the function of promoters specific to cell type. However, sensory receptor cells, particularly chemosensory cells, have been difficult to transfect. We have successfully introduced plasmids expressing green and Discosoma Red fluorescent proteins (GFP and DsRed) into rat taste buds in primary culture. Transfection efficiency increased when delaminated taste epithelium was redigested with fresh protease, suggesting that a protective barrier of extracellular matrix surrounding taste cells may normally be present. Because taste buds are heterogeneous aggregates of cells, we used α‐gustducin, neuronal cell adhesion molecule (NCAM), and neuronal ubiquitin carboxyl terminal hydrolase (PGP9.5), markers for defined subsets of mature taste cells, to demonstrate that liposome‐mediated transfection targets multiple taste cell types. After testing eight commercially available lipids, we identified one, Transfast, that is most effective on taste cells. We also demonstrate the effectiveness of two common “promiscuous” promoters and one promoter that taste cells use endogenously. These studies should permitex vivostrategies for studying development and cellular function in taste cells. © 2005 Wiley Periodicals, Inc. J. Neurobiol, 2005