Park Kiyun, Kwak Ihn-Sil
Department of Fisheries and Ocean Science, Chonnam National University, Yeosu, Korea.
Environ Health Toxicol. 2014 Jun 13;29:e2014002. doi: 10.5620/eht.2014.29.e2014002. eCollection 2014.
Heat shock protein 90 (HSP90) is a highly conserved molecular chaperone important in the maturation of a broad spectrum of protein. In this study, an HSP90 gene was isolated from Asian paddle crab, Charybdis japonica, as a bio-indicator to monitor the marine ecosystem.
This work reports the responses of C. japonica HSP90 mRNA expression to cellular stress by endocrine disrupting chemicals (EDCs), such as bisphenol A (BPA) and 4-nonylphenol (NP) using real-time. reverse transcription polymerase chain reaction.
The deduced amino acid sequence of HSP90 from C. japonica shared a high degree of homology with their homologues in other species. In a phylogenetic analysis, C. japonica HSP90 is evolutionally related with an ortholog of the other crustacean species. The expression of HSP90 gene was almost distributed in all the examined tissues of the C. japonica crab but expression levels varied among the different body parts of the crabs. We examined HSP90 mRNA expression pattern in C. japonica crabs exposed to EDCs for various exposure times. The expression of HSP90 transcripts was significantly increased in C. japonica crabs exposed to BPA and NP at different concentrations for 12, 24, 48 and 96 hours. The mRNA expression of HSP90 gene was significantly induced in a concentration- and time-dependent manner after BPA or NP exposures for 96 hours.
Taken together, expression analysis of Asian paddle crab HSP90 gene provided useful molecular information about crab responses in stress conditions and potential ways to monitor the EDCs stressors in marine environments.
热休克蛋白90(HSP90)是一种高度保守的分子伴侣,对多种蛋白质的成熟至关重要。在本研究中,从日本蟳(Charybdis japonica)中分离出一种HSP90基因,作为监测海洋生态系统的生物指标。
本研究报告了日本蟳HSP90 mRNA表达对内分泌干扰化学物质(EDCs),如双酚A(BPA)和4-壬基酚(NP)引起的细胞应激的反应,采用实时逆转录聚合酶链反应进行检测。
日本蟳HSP90推导的氨基酸序列与其在其他物种中的同源物具有高度同源性。在系统发育分析中,日本蟳HSP90与其他甲壳类物种的直系同源物在进化上相关。HSP90基因的表达几乎分布在日本蟳蟹的所有检测组织中,但在蟹的不同身体部位表达水平有所不同。我们检测了暴露于EDCs不同时间的日本蟳蟹中HSP90 mRNA的表达模式。在暴露于不同浓度的BPA和NP 12、24、48和96小时的日本蟳蟹中,HSP90转录本的表达显著增加。在暴露于BPA或NP 96小时后,HSP90基因的mRNA表达以浓度和时间依赖性方式显著诱导。
综上所述,亚洲蟳HSP90基因的表达分析提供了有关蟹在应激条件下反应的有用分子信息,以及监测海洋环境中EDCs应激源的潜在方法。