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目的 研究胚胎小鼠下丘脑培养状态下 SRIF神经元的发生。 方法 采用原代分离培养的方法 ,将胚胎小鼠下丘脑进行体外分散细胞培养 ,观察了下丘脑细胞的生长分化过程 ,并且对培养不同时间的细胞作生长抑素 (SRIF)免疫细胞化学染色 ,阳性神经元记数 ,各种形态神经元所占的百分比统计。 结果 胚胎小鼠下丘脑SRIF阳性神经元在体外生长发育不同时间细胞指数不同 :7d时为 2 4± 1.5 2 ,10 d时为 34± 2 .2 0 ,12 d时为 34±3.30 ,14d时为 10 4± 6 .32 ,16 d时为 6 8± 5 .34 ,2 0 d时为 2 9± 1.5 4,其中在培养 14d时 ,SRIF阳性神经元达到高峰 ,以后出现下降趋势。而且神经元的形态、单极、双极、多极神经元的数目以及突起呈色深浅在发育过程中也有较大的变化 ,各种占比例较大神经元出现的时期也不一样 ,其中 ,单极在培养 10、14、16 d时分别占 5 2 .3%、6 0 .0 %、43.3%。双极在培养 10、14、16 d时分别占 30 .7%、2 0 .7%、30 .1%。多极神经元在培养 7、12、2 0 d时分别占 81.5 %、90 .1%、6 7.3%。 结论 (1)在培养状态下 ,下丘脑不同核区的细胞发育速度不同 ,并且随时间点的变化呈现规律性变化 ;(2 )不同类型的 SRIF神经元的发育随着不同的时间点呈现出不同比例很可能反映一些 SRIF神经元的功能发?
Objective To study the generation of SRIF neurons under hypothalamus culture in embryonic mice. Methods Primary cultured cells were isolated and cultured in vitro. The growth and differentiation of hypothalamus cells were observed. Immunocytochemical staining of somatostatin (SRIF) was performed on cultured cells at different time points. Positive neuron count, the percentage of various morphological neurons statistics. Results The number of SRIF positive neurons in hypothalamus of embryonic mice differed at different time points in vitro: 24 ± 1.5 2 at 7 d, 34 ± 2 at 10 d, 34 ± 3.30 at 12 d and 14 ± 3 at 14 d Was 10 4 ± 6. 32, 16 8 ± 5. 34 at 16 d, and 29 ± 1.5 4 at 20 d. The expression of SRIF positive neurons peaked at 14 d and then decreased. Moreover, the morphology of neurons, the number of unipolar, bipolar, multipolar neurons and the color depth of protuberances also changed greatly during the development, and the periods of occurrence of neurons of various proportions were also different. Among them, Monopoles accounted for 52.3%, 60.0% and 43.3% respectively at 10, 14 and 16 days. Bipolar cultures accounted for 30.7%, 20.7% and 30.1% at 10, 14 and 16 days respectively. Multipolar neurons accounted for 81.5%, 90.1% and 6 7.3% respectively at 7, 12 and 20 days. Conclusions (1) Under the culture conditions, the rate of cell development in different nuclear regions of the hypothalamus is different and shows regular changes with time. (2) The development of different types of SRIF neurons appears at different time points Different proportions are likely to reflect the functional development of some SRIF neurons?