Dbn1在小鼠脑发育过程中表达与定位的研究

Expression and localization of Dbn1 in the developmental mouse brain

目的探索小鼠发育调节脑蛋白Dbn1在脑发育中的功能,检测在小鼠脑发育不同阶段的表达模式和定位。方法运用免疫组化和Western blot方法,检测小鼠脑发育不同阶段(E14,P1和P7d)以及成年(adult),Dbn1蛋白的表达模式和细胞内定位。结果Western blot方法检测小鼠各发育阶段全脑的Dbn1蛋白表达模式为:在胚胎14d时即有很高的表达,P1有所降低,P7期又有增加,成年时最低。免疫组化进一步显示:在E14时,Dbn1主要表达在大脑皮层,海马和室管膜区域,阳性信号弥散分布在细胞内;在P1期,Dbn1主要表达在海马,室管膜两侧,阳性信号在细胞内定位趋向于细胞周围及突起;P7期,Dbn1主要表达在大脑皮层,海马和神经元核团,阳性信号在细胞内的定位集中在细胞的突起;在成年2个月期,Dbn1的表达明显减弱,仅少量表达在某些神经核团,细胞内定位不十分明显。结论在小鼠发育过程中,Dbn1蛋白在脑内的表达为胚胎期最高随后降低,成年后表达甚微;而随着细胞的分化成熟,其在胞内的定位呈现出从细胞质向细胞突起集中的趋势,提示Dbn1在调节神经系统发育中有重要的作用,可能参与神经元的发育和迁移以及神经网络的建立的调节。

Objective To detect the expression changes and location of the mouse developmental regulation brain protein （ Dbn1 ） in the developmental mouse brain. Methods Monoclonal antibodies against drebrin protein were used to assess Dbn1 by immunohistochemistry and Western blot. The paternal expression of Dbn1 in developmental mouse brain （E14, P1, P7 and adult） was initially investigated by Western blot. Dbn1 was shown at various developmental stages （E14, P1 and P7） as well as in adult in different brain area of developmental mouse brain by immunohistochemical. Results Dbn1 protein was detected in developmental brain but a little in adult brain by Western blot, high at E14, decreased at P1, gradually increased at P7 and lowest in adult. Immunohistochemistry confirmed as follows： Dbn1 expressed mostly in cortex, hippocampus and ependyma areas at E14, and the positive signal was distributed at ceils border; The expression of Dbn1 was decreased at P1, mainly distributed at the verge of cells or dendrites ; The peak expression of Dbn1 appeared at P7, Dbn1 located at nucleus of hippocampus and cortex, and the positive signal located within cytoplasm and dendrites; Only a little positive Dbn1 cells were found in adult mouse brain. Conclusion Dnbl may be involved in regulating the differentiation and migration of neurons during the development of mouse brain.