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目的研究局灶性脑缺血后室管膜/室下区细胞的迁移分化,揭示梗塞区周围新生细胞的来源。方法大脑中动脉阻塞前,将10μl0.2%的荧光染料DiI注射于体质量250~350g的雄性SD大鼠侧脑室以预标记室管膜/室下区细胞。脑缺血后,采用累积式的BrdU标记方法标记新生细胞并通过双重免疫荧光染色确定细胞分化。标记的细胞通过激光共聚焦显微镜观察。结果在非缺血对照大鼠,DiI标记细胞定居于室管膜/室下区。局灶性脑缺血后,DiI标记细胞出现于胼胝体,邻近的纹状体和皮质。此外,缺血14d后,梗塞区周围纹状体和皮质内可见一些DiI/BrdU/GFAP或DiI/BrdU/NeuN三重标记阳性细胞。结论局灶性脑缺血后,室管膜/室下区细胞迁移到梗塞区周围并分化成神经元和星形胶质细胞,这一发现对于理解成体神经干细胞的起源和开发促进脑损伤后内源性神经发生的新措施具有重要意义。
Objective To study the migration and differentiation of ependymal / subventricular zone after focal cerebral ischemia, and to reveal the origins of newborn cells around the infarct zone. Methods Before occlusion of the middle cerebral artery, 10μl 0.2% fluorescent dye DiI was injected into the lateral ventricle of male SD rats with the body weight of 250 ~ 350g to pre-mark the ependymal / subventricular zone cells. After cerebral ischemia, newborn cells were labeled using a cumulative BrdU labeling method and cell differentiation was confirmed by double immunofluorescence staining. Labeled cells were observed by laser confocal microscopy. Results In non-ischemic control rats, DiI-labeled cells colonized the ependymal / subventricular zone. After focal cerebral ischemia, DiI-labeled cells appeared in the corpus callosum, adjacent striatum and cortex. In addition, some DiI / BrdU / GFAP or DiI / BrdU / NeuN triple positive cells were observed in the striatum and cortex around the infarct zone 14 days after ischemia. Conclusion After focal cerebral ischemia, the ependymal / subventricular zone cells migrate around the infarct zone and differentiate into neurons and astrocytes, a finding that is important for understanding the origin of adult neural stem cells and for promoting brain injury New measures of endogenous neurogenesis are of great importance.