大鼠穹隆海马伞切割对海马伞内神经干细胞再生的影响  

Transection of the fimbria fornix inducing neurogenesis in its proximal portion in adult rats

作　　者：高露[1] 李伟[1] 金国华[1] 张新化[1] 邹琳清[1] 田美玲[1] 秦建兵[1] 

机构地区：[1]南通大学医学院人体解剖学系,江苏省神经再生重点实验室,江苏南通226001

出　　处：《解剖学报》2011年第6期751-755,共5页Acta Anatomica Sinica

基　　金：2010年江苏省高等学校大学生实践创新训练资助项目;江苏省高校优势学科建设工程资助项目(PAPD)

摘　　要：目的检测损伤的穹隆海马伞内自体神经干细胞的再生情况。方法 SD大鼠7只,切割左侧穹隆海马伞,术后腹腔注射5-溴脱氧尿嘧啶核苷(BrdU)。术后7d取脑组织制备冷冻切片,行Nestin、BrdU免疫荧光检测;取穹隆海马伞组织进行神经干细胞的体外分离培养,对获得的细胞球进行增殖性、胚胎性以及多分化潜能的检测。结果切割侧穹隆海马伞中见到较多的Nestin和BrdU阳性细胞,部分细胞表现为BrdU/Nestin双重阳性;取切割侧穹隆海马伞组织进行分离培养能获得Nestin与BrdU阳性的神经球,而正常侧海马伞未能培养出神经球;神经球可分化为微管相关蛋白-2(MAP-2)阳性细胞、胶质纤维酸性蛋白(GFAP)阳性细胞或2,3-环核苷酸-3-磷酸二酯酶(CNP)阳性细胞。结论切割穹隆海马伞后,切割侧穹隆海马伞中出现了自体神经干细胞的再生及增殖。Objective To determine the neurogenesis in injured fimbria fornix （FiFx）. Methods The left fimbria fornix of SD rat （n = 7） were subjected to transection followed by intraperitoneal injection of 5-bromo-2-deoxyuridine （BrdU）. Coronal sections through hippocampus were stained with Nestin and BrdU antibodies 7 days after operation. The normal and injured FiFx were dissociated for neural stem cell specific culture 7 days post transection. Immunofluorescenee staining was used to determine the properties of the neurospheres. Results Immunofluorescence revealed that Nestin-expressing ceils were present in FiFx following lesion. Majority of Nestin positive cells were also positive for BrdU, a marker of DNA synthesis. Nestin positive proliferative cells were almost entirely absent from unlesioned tissue. Neurospheres cultured in vitro from lesioned FiFx displayed the characteristics of neural stem cells （NSCs） proliferation, expression of embryonic markers, and muhipotential differentiation into neurons, astrocytes and oligodendrocytes. Conclusion These results demonstrate that transection of the tracts related with hippocampal function can stimulate NSCs proliferation in the projection pathways themselves. Ectopic neurogenesis in FiFx may provide an additional mechanism for repair of innervation following damage.

关 键 词：穹隆海马伞 神经干细胞 神经元 切割 细胞培养 免疫荧光 大鼠 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]