初生大鼠前脑基底某些神经核区和视网膜的神经元在培养中相互作用的研究

STUDY OF THE INTERACTIONS BETWEEN CULTURED NEURAL CELLS FROM CERTAIN NUCLEI OF THE BASAL FOREBRAIN AND RETINA OF NEWBORN RAT

为研究早老性痴呆病时前脑基底部老年斑块和神经原纤维缠结病变发生的原因,本实验采用分离脑神经细胞体外培养和MTT微量比色法,观察了新生大鼠前脑基底的杏仁核(A)、海马(H)、隔内侧核(S)、Meynert基底核(M)和视网膜(R)细胞生长活性和形态特点,以及它们之间的相互影响。培养3天后,MTT微量比色测定发现,分别取自两核区的细胞混合培养A+H、H+R的细胞生长明显活跃,细胞活性分别比单部位培养增加212％和270％(P<0.01);A+R、A+S、H+S和S+R混合培养也均比单部位培养分别增加157％、153％、192％和158％不等(P<0.01)。另外,不同核区的条件培养液,对细胞生长活性也有促进作用,如A的条件培养液(Ac)使H细胞生长活性增加186％(P<0.05);而Sc,Rc、Hc和Mc条件培养液均能使A区细胞生长分别提高150％、144％、137％及133％(P<0.01)。在单部位培养中,A区细胞形态多为圆形,体积较大(约15μm);H区细胞体积较小(约5～7μm)圆形为主;S区细胞体积大(约10～20μm)形态不规则;M区有大小两种细胞,大的为梭形及多角形,小的多为圆形;R细胞生长不大活跃,多为小圆细胞(5～7μm)。在各混合培养与加条件培养液培养中,生长活跃与体积大的细胞增多,并长出突起;与MTT的结果一致。这些结果表明,A区细胞的生长活性增加,可能受S、R、H、M各区所分泌的生长因子促进。

In order to study the cause of mental deterioration and increase of plaques and tangles in the basal forebrain in Alzheimer's disease, we have used isolated neural cells from the nuclei of amygdala (A), the hippocampus(H), the nuclei of medial septum (S), and the basal nuclei of Meynert (M) of basal forebrain, and the retina (R) of newborn rats, to study the effect of cellular interactions on the growth activity of these cells. The growth activity was measured quantitatively using MTT colorimetric microassay method. The MTT results revealed that, after 3 days of culture, the most active cell growth were found in mixed cultures of A+H and H+R reaching up to 212％ and 270％ of their respective individual control cultures (p<0.01) and the next active cell growth were found in mixed cultures of A+R, A+S, H+S, and S+R reaching up to 157％,153％,192％ and 158％ of their respective individual control cultures (P<0.01). Further, we had observed that conditioned medium from amygdala (Ac) could activate H cell growth activity to 186％ (P<0.05).Conditioned media from medial septum(Sc), retina(Rc), hippocampus (Hc), and nuclei of Meynert (Mc) could also activate A cell growth activity to 150％,144％,137％ and 133％, respectively (P<0.01). Morphologically individual control cultures from A composed mainly of large round cells (15 μm), from H mainly of small ronnd cells (5-7 μm), from S of large and irregular cells (10-20 μm), from M of small round cells as well as large fusiform or polygonal shaped cells, and from R of inactive small round cells (5-7 μm). In mixed cultures or cultures with conditioned medium the number of active and large cells increased, some with cell processes. These results were consistent with those of MTT. These results indicated that the growth activities of A cells could be stimulated by growth factor (s)from S, R, H, and M regions. Therefore, we concluded that our study of the possible growth factor (s) from A, H, R, and S might be able to contribute to the investigation of the development and the pathogenesis of Alzheimer's disease.