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目的探讨小鼠小脑皮质发育中神经元凋亡的规律和机制。方法用激活型Caspase-3多抗免疫组织化学标记及Hoechst 33258染色液染色,检测从出生至成年小鼠小脑皮质中神经元的凋亡,用Western blotting方法对小脑组织中Caspase-3和Caspase-8的活化片段进行半定量测定。结果外颗粒层、普肯耶细胞层和内颗粒层凋亡细胞密度最高分别在出生后第8d(P8)、P5及P9,P20各层凋亡细胞密度都很低。Caspase-3活化片段的表达量在P5较高,P5以后逐渐降低,至P14消失;Caspase-8活化片段的表达量从P0到P10都较高,P10以后逐渐降低,至P30消失。结论P0至P14是小脑皮质神经元凋亡的重要时期,通过启动Caspase-8的活化进而激活效应性Caspase-3的细胞凋亡途径存在于小脑皮质的塑型成熟过程。
Objective To investigate the regularity and mechanism of neuronal apoptosis during cerebellar cortex development in mice. Methods The apoptotic neurons in the cerebellar cortex from birth to adult mice were detected by activated Caspase-3 polyclonal antibody immunohistochemistry and Hoechst 33258 staining. The expressions of Caspase-3 and Caspase-3 in cerebellum were detected by Western blotting. 8 activation fragment for semi-quantitative determination. Results The density of apoptotic cells in the outer granular layer, the Pukeye layer and the inner granular layer were the lowest at 8d (P8), P5, P9 and P20 layers after birth, respectively. The expression of Caspase-3 activation fragment was higher in P5, gradually decreased after P5, disappeared to P14; the expression of Caspase-8 activation fragment was higher from P0 to P10, then decreased gradually after P10, disappeared to P30. Conclusion P0 to P14 is an important period of cerebellar cortical neuron apoptosis. Activation of Caspase-8 activates the apoptotic pathway of effector Caspase-3 in the process of plastic maturation of cerebellar cortex.