流式细胞计用于大鼠脑发育研究

A DEVELOPMENTAL STUDY OF RAT BRAIN BY FLOW CYTOMETRY

用胚胎１６ｄ（ｅｍｂｒｙｏｎｉｃｄａｙ１６，Ｅ１６）、Ｅ１８、Ｅ２０、新生和成年Ｗｉｓｔａｒ大鼠各５只，取大脑、小脑、中脑、脑桥、延髓和脊髓组织制成细胞悬液，细胞悬液中，绝大部分细胞变为圆形，少数有突起的细胞经２—４ｈ的悬液培养突起回缩，细胞变圆后可顺利通过流式细胞计（ｆｌｏｗｃｙｔｏｍｅｔｒｙ，ＦＣＭ）上直径７０μｍ的喷嘴，证实ＦＣＭ可用于神经科学的研究，用ＦＣＭ测得Ｅ１６大鼠１万个细胞中Ｇ２／Ｍ期细胞数大脑明显高于小脑，Ｅ１８时大、小脑Ｇ２／Ｍ期细胞数达高峰，此时二者间无显著差异；Ｅ１６、Ｅ１８、Ｅ２０，中脑、脑桥、延髓和脊髓１万个细胞中Ｇ２／Ｍ期细胞数明显低于大、小脑。新生大鼠脑各部Ｇ２／Ｍ期细胞数急剧降低，成年与新生比无显著差异。

By using flow cytometry (FCM), 75 Wistar rats from different developmental stage (E16,E18, E20, neonate and adult) were studied. Suspensions were maken from cerebra, cerebella, mesencephalons,poutes, medulla and spinal cords respectively in which most of the cells had become round in shape and the rest withdrawed their processes after a culture of 2-4 Hrs. Thus the brain cells could go through the nozzlo (Φ: 70 μm) of FCM and be sandied. In the present study, FCM had been used tO measure the cell number of G2/ M phase in every 10 000 brain cells of various regions (mentioned above) from E16, E18, E20, neonatal and adult rats. The results showed that the G2/ M cells were significantly more in cerebrum and cerebellum than in mesencephalon, pons, medulla and spinal cord in E16, E18 AND E20. It decreased dramatically in all brain areas stodied in neonates. No significant difference was found in G2/ M cells between neonate and adult. G2/ Mcells were significantly higher in cerebrum than in cerebrllum in E16 and reached the hightest level in both cerebrum and cerebellum in E18 with no significant difference between them.