大鼠中脑黑质器官型脑片与在体发育黑质结构的形态学比较

Comparison on the Morphology of the Organotypic Brain Slice Culture in Vitro of Substantia Nigra and in Vivo

目的:比较不同时间离体培养黑质脑片与在体发育黑质结构的形态学变化。方法:选出生当天SD大鼠,制备中脑黑质器官型脑片(厚度350μm)。用Millicell-CM微孔膜脑片离体培养技术分别培养脑片至7、14、21和28 d,光镜观察,常规苏木精-伊红染色及酪氨酸羟化酶(TH)免疫组化反应染色,并与同龄SD大鼠作形态学及TH阳性反应细胞计数比较。结果:培养0～28 d内器官型脑片黑质细胞的体积及细胞密集程度均小于同期在体发育的黑质结构;离体培养与同期在体发育的黑质结构比较,TH阳性反应细胞趋于幼稚,细胞间连接也出现较晚,两者细胞计数比较有统计学意义(P<0.05)。结论:SD大鼠黑质发育的成熟时间约为生后14 d;离体培养的黑质脑片与在体生长的黑质结构发育过程相似,但离体培养的黑质脑片发育滞后于在体发育的黑质结构。

Aim ： To compare the morphology change of the organotypic brain slice culture of substantia nigra and in different time in vivo. Methods. The organotypic brain slice culture of substantia nigra prepared from neonatal SD rats were placed on Millicell-CM porous membranes and cultured to day-7, day-14, day- 21 and day-28. The HE staining with stereological methods and tyrosine hydroxylase （TH） immunohistochemical staining with the cell counting were used to compare the brain slice culture with the same age rats. Results ： The substantia nigra cells cultured from 0 to 28d were smaller and the density of which were lower than that of in vivo of the same age. TH^＋ cells cultured were more immature than those in vivo, and the occurrence of cell junctions was later, between which there has been significant difference（P〈0.05） Conclusion ： The maturate time of substantia nigra in SD rats was approximately 14 d after neogenesis. The process of substantia nigra development in the organotypic brain slice culture in vitro was similar to that in vivo, but the development in vitro was hysteretic behind the postnatal development in vivo.