巢蛋白在胚胎及新生大鼠脊髓的表达

The Expression of Nestin in Spinal Cord of Rat from Embryo to Newborn

目的 检测不同发育时期巢蛋白 (nestin)在脊髓神经干细胞的表达水平 ,探讨脊髓神经干细胞的发育规律 ,为分离、富集和增殖均质的神经干细胞以及后期的研究工作如神经干细胞定向分化及移植治疗等研究提供实验依据。方法 将Wistar大鼠按胎龄及日龄分组 ,剥离脊髓 ,制成单细胞悬液 ,经间接荧光法标记nestin抗原 ,用流式细胞仪检测和分析。结果 从E13到P7整个发育过程中都有nestin抗原的表达 ,呈升高趋势 ,其中P1期表达水平最高。出生后表达下降 ,趋近于 90d(同型对照 )表达水平。结论 巢蛋白在脊髓的表达出现于胚胎早期 ,表达水平随胎龄轻微波动 ,呈升高趋势 ,P1达到高峰 ,出生后表达下降 。

Objective To find the effective strategies of isolation, enriching and proliferation of homogeneous neural stem cells (NSCs) in vitro, we detected the expression of nestin positive cells in spinal cord at different stages of development in rat to contribute to the further understanding of the neurogenesis of spinal cord. Method Wistar rats were grouped into nine parts according to different stages of development, such as E13, E15, E17, E19, P1, P3, P5, P7, P90 (serving as control). Spinal cords were microsurgically obtained and made into suspensions. Nestin antigens of NSCs in the suspensions were combined with monoclonal antibody of nestin, then the compounds were fixed with the second antibodies of IgG FITC, which were determined with flow cytometry. Results Nestin antigens were expressed during the investigated period with a trend of slight fluctuation from E13 to P1 with the highest level in P1, then drastically decreased until P7 with the same level as that of 90d. Conclusion The expression of nestin in the spinal cord begins at early stage of rat embryo, accompanied with slightly fluctuateion, and reached the summit at P1, then decreased after born and tended to be the level of adult within a postnatal week.