锌铁转运蛋白9在睾酮调节小鼠海马突触可塑性过程中的作用  

The role of Zrt-and Irt-like protein 9 on testosterone induced hippocampal synaptic plasticity in mouse

作　　者：张沂洲 李莎[1] 左洪春 宋磊刚 杜鹃[1] 王畅[1] 崔慧先[1] Zhang Yizhou;Li Sha;Zuo Hongchun;Song Leigang;Du Juan;Wang Chang;Cui Huixian(Department of Human Anatomy,Hebei Key Laboratory of Neurodegenerative Disease Mechanism,Hebei Medical University,Shijiazhuang 050017,China)

机构地区：[1]河北医科大学人体解剖学教研室,河北省神经退行性疾病机制研究重点实验室,石家庄050017

出　　处：《神经解剖学杂志》2021年第6期623-629,共7页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金(91849134);河北省自然科学基金(C2020206009);河北省高等学校科学技术研究项目(QN2020131)。

摘　　要：目的:研究睾酮通过锌铁转运蛋白9(ZIP9)对海马CA1区神经元突触后致密蛋白95(PSD95)表达的非基因组效应及其作用机制.方法:腺相关病毒感染3月龄C57BL/6小鼠海马CA1区神经元,敲低锌铁转运蛋白9(ZIP9)表达,并通过Western Blot实验检测敲低效率;Golgi-cox染色实验观察去势手术或ZIP9-RNAi预处理后膜不通透性类固醇偶联物睾酮-胎牛血清白蛋白(T-BSA)对小鼠海马CA1区神经元树突棘密度的影响;Western Blot实验观察去势手术或ZIP9-RNAi预处理后,T-BSA对小鼠海马CA1区神经元PSD95蛋白、细胞外调节蛋白激酶1/2(Erk1/2)磷酸化和总蛋白、真核起始因子4E(eIF4E)磷酸化和总蛋白水平的影响.结果:海马CA1区可见病毒感染后绿色荧光GFP标记神经元,Western Blot结果显示ZIP9-RNAi预处理后海马CA1区ZIP9蛋白表达显著降低;Golgi-cox染色实验结果显示去势手术后小鼠海马CA1区神经元树突棘密度显著降低,T-BSA可抑制去势手术后树突棘密度的降低,而ZIP9-RNAi预处理显著抑制T-BSA对小鼠海马CA1区神经元树突棘密度的恢复作用;Western Blot实验结果显示去势手术后小鼠海马CA1区PSD95蛋白水平、Erk1/2和eIF4E磷酸化蛋白水平显著降低,T-BSA可恢复去势手术后上述蛋白的降低,而ZIP9-RNAi预处理显著抑制T-BSA对上述蛋白的恢复作用,但对Erk1/2和eIF4E总蛋白水平未见显著影响.结论:T-BSA可通过ZIP9介导的Erk1/2-eIF4E信号通路影响PSD95蛋白的表达,进而影响海马神经元突触可塑性.Objective: To study the non-genomic effect of testosterone on the postsynaptic density protein 95(PSD95) protein expression via Zrt-and Irt-like protein 9(ZIP9) in hippocampal neurons and the mechanism of this action. Methods: Adeno associated virus infected neurons in hippocampal CA1 region of 3-month-old C57 BL/6 mice to knock down the expression of ZIP9, and the knockdown efficiency was detected by Western Blot. The effects of membrane-impermeable steroid T-BSA on dendritic spine density of hippocampal CA1 neurons after pretreatment with castration or ZIP9-RNAi were observed by Golgi-cox staining. The effects of T-BSA on PSD95, p-Erk1/2, p-eIF4 E, t-Erk1/2, and t-eIF4 E protein levels of hippocampal CA1 after pretreatment with castration or ZIP9-RNAi were observed by Western Blot. Results: Green fluorescent GFP labeled neurons were observed in hippocampal CA1 region after virus infection. Western Blot results showed that ZIP9-RNAi pretreatment significantly reduced the ZIP9 protein level. Golgi-cox staining results showed that dendritic spine density of hippocampal CA1 decreased significantly after castration, and T-BSA rescued it, while ZIP9-RNAi pretreatment significantly inhibited the corrective effect of T-BSA on dendritic spine density. Western Blot showed that the levels of PSD95, p-Erk1/2, and p-eIF4 E protein in hippocampal CA1 were significantly decreased after castration, and T-BSA rescued these effects. But the effect of T-BSA on the above-mentioned proteins in hippocampal CA1 were significantly inhibited after pretreatment with ZIP9-RNAi. There was no significant effect on t-Erk1/2 and t-eIF4 E protein levels. Conclusion: T-BSA affects the expression of PSD95 through Erk1/2-eIF4 E signaling pathway mediated by ZIP9, and then affects the synaptic plasticity of hippocampal neurons.

关 键 词：睾酮 非基因组效应 锌铁转运蛋白9 突触后致密蛋白95 小鼠 

分 类 号：R285.5[医药卫生—中药学]