内源性神经保护蛋白Iduna在新生大鼠脑组织中的表达及定位

Expression and localization of endogenous neuroprotective protein Iduna in neonatal rat brain

目的明确内源性神经保护蛋白Iduna在新生大鼠脑组织中的表达水平及定位特点。方法取SD新生鼠各器官组织和脑分区,用Western blot评价Iduna蛋白表达的组织特异性;制作SD新生鼠脑组织石蜡与冰冻切片,免疫组织化学观察Iduna蛋白的表达及定位特点;免疫荧光三标脑组织冰冻切片确认Iduna在神经元及星形胶质细胞中的定位;培养原代神经元及星形胶质细胞,real-time PCR定量分析Iduna mRNA在细胞中的表达和免疫荧光双染Neu N和GFAP验证Iduna的细胞定位。结果 Iduna在新生鼠脑组织中表达丰富,尤其是海马和皮层区域。Iduna主要定位在大脑海马神经元细胞的胞质内(79.85%),原代神经元中也验证了这一结果(80.6%)。Iduna mRNA在神经元中的表达丰度远远高于星形胶质细胞(P<0.001)。结论脑组织中Iduna主要分布在海马和皮层区域的神经元胞质中,提示其功能可能与神经元的活动密切相关。

Objective To clear the expression of endogenous neuroprotective protein Iduna in brain tissues of neonatal rats and the localization. Methods The organ tissues and different brain regions of SD neonatal rats were taken for evaluating the tissue specificity of Iduna protein expression using Western blot. The brain paraffin and frozen section of SD neonatal rats were made. Then to observe the expression of Iduna and the localization through IHC,and determine the positioning of Iduna in nerve cell and astrocyte through IF. Culture the primary neurons and astrocyte,and detect the expression of Iduna mRNA through the real-time PCR. Verify the cellular localization of Iduna through the IF double-stained Neu N and GFAP. Results Iduna had a rich expression in the brain of neonatal rats,especially in hippocampus and cortex. Iduna was mainly expressed in the cytoplasm of neuron（ 79. 85%）,that was also proved by primary neurons IF triple-staining（ 80. 6%）. The expressive abundance of Iduna mRNA in neurons was much higher than that in astrocytes（ P 〈0. 001）. Conclusions Iduna is mainly distributed in the neuron plasma in hippocampus and cortex of brain,indicating that the function may be closely related to neuron＇s activity.