目的:研究Sytnaptotagmin(Syt)I和Ⅳ在LβT2细胞中的定位和shRNA干扰技术对这两种蛋白在促性腺激素释放细胞(LβT2)细胞中蛋白表达量的影响。方法:运用全内反射荧光显微系统,通过在LβT2细胞中共表达各种细胞器的蛋白标记物和Syt I、Ⅳ与荧光蛋白的融合蛋白,对SytI、Ⅳ进行细胞内定位。并运用shRNA干扰技术对sytI、Ⅳ在L8T2细胞中蛋白表达量进行了干扰。结果:发现了sytI、sytⅣ在LβT2细胞中和致密核心大囊泡(LDCV)都存在了一定的共存比例,而且shRNA技术对SytI和SytⅣ在LβT2细胞中的表达沉默产生了一定的效用。结论:SytI、SytⅣ在LβT2细胞中与致密核心大囊泡共存比例很大,shRNA干扰技术对SytI、SytⅣ的沉默具有高效性,为未来进一步对SytI、SytⅣ在LβT2细胞中的功能研究打下了基础。
Objective: To observe localizations of Syt I, and Syt IV in LβT2 cells and test the expression of Syt I, IV in LβT2 cells after silenced by shRNA. Methods: We used the total internal reflection fluorescence microscopy(TIRFM) to observe localizations of Syt I, and Syt IV in LβT2 cells, and silenced the expression Syt I,IV by shRNA technology. Results: The fusion proteins of Syt I, IV with fluorescent proteins are mostly co-localized with the maker of the large dense vesicle. And shRNA technology has a good efficiency on silencing Syt I, Syt IV in LβT2 cells. Conclusions: Syt I, Syt IV are colocalized with LDCV in LβT2 cells. The technology of shRNA for Syt I and Syt IV has a good efficiency and it is useful for the Syts function research in LβT2 cells.