β-半乳糖苷酶在ROSAβgeo26小鼠的脑和神经干细胞中的表达

发布时间：2018-08-29 12:13

【摘要】：ROSAβgeo26(ROSA26)小鼠线系是由胚胎干细胞线系衍生而来，其在胚胎发育过程中的所有细胞均表达报告基因LacZ，故被视为骨髓移植实验中的一个有价值的标记物线系。LacZ基因的产物β-半乳糖苷酶在ROSA26小鼠线系的全身组织广泛表达。然而，，β-半乳糖苷酶在该小鼠线系脑中的细胞分布和转基因神经干细胞在移植中的利用还有待于研究。为此，本实验采用X-Gal染色、尼氏染色和免疫组织及细胞化学染色方法研究了β-半乳糖苷酶在ROSA26小鼠的脑和神经干细胞中的表达特征。结果显示：β-半乳糖苷酶在新生和成年ROSA26小鼠脑的神经干细胞和神经元中广泛表达，而在星形胶质细胞中的表达下调。转基因神经干细胞和星形胶质细胞的培养实验进一步证实了在体实验的研究结果。移植实验显示整合到宿主大鼠脑中的转基因神经干细胞在分化后的神经元具有β-半乳糖苷酶活性，而星形胶质细胞则不表达β-半乳糖苷酶的活性。上述结果提示ROSA26小鼠中的LacZ基因在神经干细胞分化为胶质细胞系的过程中沉默，故在移植研究中评价转基因细胞在宿主体内命运时应注意此现象。
[Abstract]:ROSA 尾 geo26 (ROSA26) mouse line is derived from embryonic stem cell line. The reporter gene LacZ, was expressed in all the cells during embryonic development, so it was regarded as a valuable marker line of bone marrow transplantation. The product of Lacz gene 尾 -galactosidase was widely expressed in the whole body of ROSA26 mice. However, the cell distribution of 尾 -galactosidase in the mouse linear brain and the use of transgenic neural stem cells in transplantation need to be studied. Therefore, the expression of 尾 -galactosidase in the brain and neural stem cells of ROSA26 mice was studied by X-Gal staining, Nissl staining, immunohistochemistry and cytochemical staining. The results showed that 尾 -galactosidase was widely expressed in neural stem cells and neurons of newborn and adult ROSA26 mice, but down-regulated in astrocytes. Transgenic neural stem cells and astrocytes culture experiments further confirmed the results of in vivo experiments. Transplantation experiments showed that the differentiated neurons of transgenic neural stem cells integrated into host rat brain had 尾 -galactosidase activity, while astrocytes did not express 尾 -galactosidase activity. These results suggest that the LacZ gene in ROSA26 mice is silenced during the differentiation of neural stem cells into glial cell lines, so we should pay attention to this phenomenon when evaluating the fate of transgenic cells in vivo.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R329

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