解剖学报

解剖学报 ›› 2013, Vol. 44 ›› Issue (1 ): 19-24.doi: 10.3969/j.issn.0529-1356.2013.01.004

• 神经生物学 • 上一篇    下一篇

成年大鼠最后区神经干细胞的检测及鉴定

李力，马常升，李岳，郝庆卯*，曹翠丽，张智琴，王立涛    

  1. 河北医科大学神经生物学研究室，石家庄 050017
  • 收稿日期:2012-03-28 修回日期:2012-06-15 出版日期:2013-02-06 发布日期:2013-02-06
  • 通讯作者: 郝庆卯 E-mail:hqm6609@yahoo.com.cn
  • 基金资助:

    河北省卫生厅医学科学研究重点资助课题（20100032）

Detection and identification of neural stem cells in area postrema of adult rats

LI Li,MA Chang-sheng,LI Yue,HAO Qing-mao*,CAO Cui-li,ZHANG Zhi-qin,WANG Li-tao   

  1. Laboratory of Neurological Biology, Hebei Medical University, Shijiazhuang 050017, China
  • Received:2012-03-28 Revised:2012-06-15 Online:2013-02-06 Published:2013-02-06

摘要：

目的 观察性别决定基因高迁移率组蛋白( SOX2)和巢蛋白(Nestin)阳性表达细胞及溴脱氧尿密啶核苷（BrdU）阳性标记细胞在最后区的分布。方法 成年雄性SD大鼠12只，6~8周龄，随机分为两组，每组6只。一组大鼠按照50mg/kg（0.3ml）腹腔注射BrdU，连续3d给药，每天2次；另一组注射等量生理盐水。4d后灌注大鼠，行免疫组织化学及免疫荧光检测。 结果 免疫组织化学染色显示，SOX2阳性表达细胞在最后区的腹侧部呈明显的V字形分布，背侧部呈带状分布，中央部散在分布。Nestin阳性表达细胞在最后区的腹侧部呈明显的V字形强阳性分布，中央部呈弱阳性表达。在最后区可见少量阳性BrdU标记细胞。SOX2/BrdU荧光双标染色显示，SOX2阳性表达细胞较密集分布，可见少量SOX2/BrdU双阳性细胞。SOX2/Nestin荧光双标染色显示，SOX2阳性表达细胞较密集分布，可见少量SOX2/Nestin双阳性表达细胞。结论 最后区SOX2及Nestin阳性细胞密集表达，呈明显的区域分布；在最后区内存在少量BrdU阳性标记细胞及SOX2/BrdU和SOX2/Nestin双阳性细胞；最后区可能存在神经干细胞/祖细胞。

关键词: 神经干细胞, 最后区, 性别决定基因高迁移率组蛋白, 巢蛋白, 免疫组织化学, 免疫荧光, 大鼠

Abstract:

Objective To observe the expression and distribution of SRY-related high-mobility-group-box-protein 2(SOX2), Nestin and BrdU positive cells in area postrema. Methods A total of 12 healthy 6-8 weeks old male SD rats were used in this study, 6 of which were treated with intraperitoneal injections of bromodeoxyuridine(BrdU), 50 mg/kg（0.3ml), two times a day for three days and other 6 were used as the control and treated with saline. After conventional heart perfusion, area postrema was detected by immunohistochemical staining and immunofluorescence staining for SOX2, Nestin and BrdU positive cells. Results Immunohistochemical staining indicated that SOX2 positive cells showed V shape distribution in the ventral part of area postrema. On the subpial of the back side, the dense cells were found with a zonal distribution. In the central part of area postrema, the positive cells were scattered and stained lighter. Nestin positive cells in the ventral part of area postrema showed V shape strongly positive color. Nestin showed weakly positive color in the central department of area postrema. The area postrema was visible with a small amount of BrdU positive cells. The SOX2 positive cells of area postrema were concentrated and shown the shape of the letter V in the ventral part. The cells were the divergence in expression in the central part. The visible positive expression of Nestin cells showed the shape of V distribution in the area postrema. Its positive immunoreactive products were granular and filaments. Immunofluorescence double staining demonstrated that in the SOX2 /BrdU fluorescence double staining specimen, SOX2 positive cells were densely distributed. Among the dense SOX2 positive cells, a small amount of SOX2/BrdU positive cells were scattered. In SOX2/Nestin fluorescence double staining specimens, SOX2 positive cells were densely distributed. The scattered distribution of SOX2/Nestin positive cells was seen in dense SOX2 positive cells. Conclusion SOX2 and Nestin positive cells show concentrated expression in area postrema. The positive cells in the ventral part show the shape of V dense distribution. BrdU labeled cells and SOX2/BrdU and SOX2/Nestin double labeled cells are visible in area postrema. These may support the existence of neural stem/progenitor cells in area postrema. 

Key words: Neural stem cell, Area postrema, SRY-related high-mobility-group-box-protein2, Nestin, Immunohistochemistry, Immunofluorescence, Rat