摘要
目的探讨体外原代培养胎鼠大脑皮层神经元NMDAR1（NR1）亚基表达的发育性变化。方法利用体外原代培养Wistar孕14～15d胎鼠大脑皮层神经元，采用免疫荧光法鉴定神经元的纯度和NR1亚基在神经元上的定位，采用流式细胞术和免疫印迹法（Western blot）检测不同培养时间神经元NR1亚基的表达情况。蛄杲体外原代培养的胎鼠大脑皮层神经元纯度均达到90％以上，免疫荧光标记显示NR1亚基主要分布在神经元胞膜及树突干膜上，流式细胞仪检测培养1、2、3、4、6、9、12d神经元NR1亚基平均荧光强度分别为4．57±0．99、8．18±1．22、13．67±1．99、20．33±1．03、26．30±2.88、31．71±2．47、28．63±1．40，westem blot显示细胞膜蛋白中NR1亚基在1～2d表达微弱，3～6d表达逐渐增高，6d以后保持稳定。结论体外原代培养的胎鼠大脑皮层神经元NR1亚基有发育性变化，这种变化与神经元的成熟度有关。
Objective To investigate developmental expression of NMDAR1 （NR1） subunit on cultured cerebral cortical neurons of fetal rats in vitro. Methods Cerebral cortical neurons were primarily cultured from pregnant 14 - 15 d fetal rats. Purity of neurons and location of NR1 subunit were identified by flow cytometry and Western blot. Results , and expression of NR1 subunit was determined by The purity of neurons was more than 90%. Immunofluoreseenee showed that NR1 subunit is mainly located on the membrane of neuronic bodies and dendritic stems. Flow cytometry showed that average fluorescence intensity of NR1 subunit on 1 d, 2d, 3d, 4d, 6d, 9d, and 12 d was 4.57± 0.99, 8.18±1.22, 13.67±1.99, 20.33 ± 1.03, 26.30 ± 2.88, 31.71 ± 2.47, and 28.63 ± 1.40, respectively. Western blot showed that the membrane protein NR1 subunit expression was weak on 1-2 d, and gradually strengthened on 3-6 d, then renmined stably. Conclusion Expression of NR1 subunit on cultured cerebral cortical neurons of fetal rats has developmental changes, which is related to neuronic maturity.
出处
《山东大学学报（医学版）》
CAS
北大核心
2009年第6期28-32,共5页
Journal of Shandong University:Health Sciences
