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紫外线B辐射暴露对淡水虾胚胎中与细胞凋亡相关的基因和蛋白质表达的影响。

Effect of UVB radiation exposure in the expression of genes and proteins related to apoptosis in freshwater prawn embryos.

作者信息

Schramm Heloísa, Jaramillo Michael L, Quadros Thaline de, Zeni Eliane C, Müller Yara M R, Ammar Dib, Nazari Evelise M

机构信息

Departamento de Biologia Celular, Embriologia e Genética, Universidade Federal de Santa Catarina, Florianópolis, Santa Catarina, Brazil.

Departamento de Biologia Celular, Embriologia e Genética, Universidade Federal de Santa Catarina, Florianópolis, Santa Catarina, Brazil; Centro Universitário Católica de Santa Catarina, Joinville, Santa Catarina, Brazil.

出版信息

Aquat Toxicol. 2017 Oct;191:25-33. doi: 10.1016/j.aquatox.2017.07.014. Epub 2017 Jul 26.

Abstract

Our previous studies showed that embryos of the freshwater prawn Macrobrachium olfersii exposed to ultraviolet B (UVB) radiation exhibited DNA damage, excessive ROS production, mitochondrial dysfunction and increased hsp70 expression, which are able, independently or together, to induce apoptosis. Thus, we attempted to elucidate some key apoptosis-related genes (ARG) and apoptosis-related proteins (ARP) and their expression during different stages of embryonic development, as well as to characterize the chronology of ARG expression and ARP contents after UVB radiation insult. We demonstrate that p53, Bax and Caspase3 genes are active in the embryonic cells at early embryonic developmental stages, and that the Bcl2 gene is active from the mid-embryonic stage. After UVB radiation exposure, we found an increase in ARP such as p53 and Bak after 3h of exposure. Moreover, an increase in ARG transcript levels for p53, Bax, Bcl2 and Caspase3 was observed at 6h after UVB exposure. Then, after 12h of UVB radiation exposure, an increase in Caspase3 gene expression and protein was observed, concomitantly with an increased number of apoptotic cells. Our data reveal that ARG and ARP are developmentally regulated in embryonic cells of M. olfersii and that UVB radiation causes apoptosis after 12h of exposure. Overall, we demonstrate that embryonic cells of M. olfersii are able to active the cell machinery against environmental changes, such as increased incidence of UVB radiation in aquatic ecosystems.

摘要

我们之前的研究表明,暴露于紫外线B(UVB)辐射下的淡水虾罗氏沼虾胚胎表现出DNA损伤、过量的活性氧产生、线粒体功能障碍以及热休克蛋白70(hsp70)表达增加,这些因素能够单独或共同诱导细胞凋亡。因此,我们试图阐明一些关键的凋亡相关基因(ARG)和凋亡相关蛋白(ARP)及其在胚胎发育不同阶段的表达情况,以及表征UVB辐射损伤后ARG表达和ARP含量的时间顺序。我们证明,p53、Bax和Caspase3基因在胚胎发育早期的胚胎细胞中具有活性,而Bcl2基因在胚胎发育中期开始具有活性。在暴露于UVB辐射后,我们发现暴露3小时后p53和Bak等ARP增加。此外,在UVB暴露6小时后,观察到p53、Bax、Bcl2和Caspase3的ARG转录水平增加。然后,在UVB辐射暴露12小时后,观察到Caspase3基因表达和蛋白增加,同时凋亡细胞数量增加。我们的数据表明,ARG和ARP在罗氏沼虾胚胎细胞中受到发育调控,并且UVB辐射在暴露12小时后导致细胞凋亡。总体而言,我们证明罗氏沼虾的胚胎细胞能够激活细胞机制以应对环境变化,例如水生生态系统中UVB辐射发生率的增加。

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