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Expression and Cell Distribution of SENP3 in Brain Tissue After Traumatic Brain Injury in Mice: A Pilot Study

机译:小鼠脑外伤后脑组织中SENP3的表达和细胞分布:一项初步研究

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SUMO-specific proteases 3 (SENP3) is a member of the small ubiquitin-like modifier-specific protease family and deconjugates SUMO2/3 from protein substrates. To date, the expression and function of SENP3 in traumatic brain injury (TBI) are unclear. The present study examined dynamic changes in SENP3 expression in the cerebral cortex and in its cellular localization, using an acute TBI model in adult mice. SENP3 expression was examined at 3, 6, 12, 24 h, 3, and 5 days after TBI using Western Blot analysis and quantitative real-time PCR. Immunohistochemistry and immunofluorescence were examined to detect SENP3 localization. Western Blot indicated that SENP3 protein levels gradually increased from 3 h after TBI and peaked at 24 h. Quantitative real-time PCR demonstrated a gradual increase in SENP3 expression, which peaked 12 h after TBI and declined subsequently. Immunohistochemical staining demonstrated that SENP3-positive cells were observed in both the sham and 24 h post-TBI groups. However, robust expression of SENP3 was seldom observed in the sham group, while it was notably enhanced after TBI. Furthermore, immunofluorescence results revealed that the expression of SENP3 increased more significantly in neurons at day 1 after TBI compared with sham group and less significantly in astrocytes and microglia. Moreover, the SENP3-positive cells that were co-expressed with NeuN also expressed caspase-3, indicating a potential correlation between SENP3 and apoptosis after TBI. Collectively, our results showed obvious up-regulation of SENP3 expression in the brain after TBI, especially in the neurons. However, the full role of SENP3 and its therapeutic potential in TBI needs further investigation.
机译:SUMO特异性蛋白酶3(SENP3)是小的泛素样修饰剂特异性蛋白酶家族的成员,可从蛋白质底物中解偶联SUMO2 / 3。到目前为止,尚不清楚SENP3在脑外伤(TBI)中的表达和功能。本研究使用成年小鼠的急性TBI模型检查了大脑皮层及其细胞定位中SENP3表达的动态变化。使用Western Blot分析和定量实时PCR在TBI后3、6、12、24 h,3和5天检查SENP3表达。检查了免疫组织化学和免疫荧光以检测SENP3的定位。 Western Blot表明SENP3蛋白水平从TBI后3小时开始逐渐升高,并在24 h达到峰值。实时定量PCR证明SENP3表达逐渐增加,在TBI后12小时达到峰值,随后下降。免疫组织化学染色表明,假手术组和TBI术后24小时均观察到SENP3阳性细胞。然而,在假手术组中很少观察到SENP3的稳定表达,而在TBI后却明显增强。此外,免疫荧光结果显示,与假手术组相比,TBI后第1天SENP3的表达在神经元中增加更为明显,而在星形胶质细胞和小胶质细胞中则不明显。此外,与NeuN共表达的SENP3阳性细胞也表达caspase-3,表明TBI后SENP3与细胞凋亡之间存在潜在的相关性。总体而言,我们的研究结果显示,TBI后脑部特别是神经元中SENP3表达明显上调。但是,SENP3的全部作用及其在TBI中的治疗潜力有待进一步研究。

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