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目的:探讨以人胶质纤维酸性蛋白(hGfap)基因启动子介导的Cre重组酶在小鼠小脑中的表达。方法:将hGfapcre转基因小鼠与Rosa26R转基因鼠杂交得到hGfapcre/Rosa26R基因型小鼠,分别在胚胎12.5、13.5、14.5、16.5 d和出生后3周,取小脑组织切片行X-gal染色观察Cre重组酶分布;另外,出生后3周的小脑组织切片同时使用细胞种类特异性抗体Blbp(星形胶质细胞标记物)、NeuN(颗粒细胞标记物)、Calbindin(浦肯野细胞标记物)进行免疫组织化学染色鉴定X-gal染色阳性细胞类型。结果:胚胎13.5 d,小脑菱唇开始出现X-gal染色阳性细胞;此后Cre重组酶表达范围进一步扩大,至胚胎16.5 d,X-gal染色阳性细胞可覆盖整个小脑;在出生后3周,X-gal染色阳性细胞可以和Blbp及NeuN共标记,和Cal-bindin不能共标记。结论:以hGfap基因启动子介导的Cre重组酶最早在胚胎13.5 d的菱唇开始表达,并且Cre重组酶主要表达于星形胶质细胞(包括Bergmann胶质细胞)和颗粒细胞,不表达于浦肯野细胞,表明在小脑发育过程中以GFAP为标记物的胶质细胞主要向星形胶质细胞(包括Bergmann胶质细胞)和颗粒细胞分化,不向浦肯野细胞分化。
AIM: To investigate the expression of Cre recombinase in mouse cerebellum mediated by hGfap gene promoter. Methods: The hGfapcre / Rosa26R genotype mice were obtained by crossing hGfapcre transgenic mice with Rosa26R transgenic mice. The recombination of Cre was observed by X-gal staining at 12.5, 13.5, 14.5, 16.5 days and 3 weeks after birth respectively In addition, cerebellum sections of 3 weeks after birth were also immunized with cell-type specific antibodies Blbp (astrocyte marker), NeuN (granulocyte marker), Calbindin (Purkinje cell marker) Histochemical staining identified X-gal staining positive cell types. Results: At 13.5 days after embryo implantation, X-gal positive cells began to appear in the rhizome of the cerebellum. After that, the expression of Cre recombinase further expanded to 16.5 days after implantation. X-gal positive cells covered the whole cerebellum. At 3 weeks after birth, -gal positive cells can be co-labeled with Blbp and NeuN, and Cal-bindin can not be co-labeled. CONCLUSION: The Cre recombinase mediated by the hGfap promoter was first expressed in the rhizome of embryonic day 13.5, and Cre was mainly expressed in astrocytes (including Bergmann glia) and granulosa cells, but not in Purkinje cells indicate that GFAP-labeled glial cells mainly differentiate into astrocytes (including Bergmann glia) and granulosa cells during cerebellar development and do not differentiate into Purkinje cells.