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Cyst viability and stress tolerance upon heat shock protein 70 knockdown in the brine shrimp Artemia franciscana

机译:盐水虾卤虫中热休克蛋白 70 敲低后的囊肿活力和应激耐受性

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Females of the brine shrimp Artemia franciscana produce either free-swimming nauplii via ovoviviparous pathway of reproduction or encysted embryos, known as cysts, via oviparous pathway, in which biological processes are arrested. While previous study has shown a crucial role of ATP-dependent molecular chaperone, heat shock protein 70 (Hsp70) in protecting A. franciscana nauplii against various abiotic and abiotic stressors, the function of this protein in diapausing embryos and cyst development, however, remains unknown. RNA interference (RNAi) was applied in this study to examine the role of Hsp70 in cyst development and stress tolerance, with the latter performed by desiccation and freezing, a common method used for diapause termination in Artemia cysts. Hsp70 knockdown was apparent in cysts released from females that were injected with Hsp70 dsRNA. The loss of Hsp70 affected neither the development nor morphology of the cysts. The time between fertilization and cyst release from Artemia females injected with Hsp70 dsRNA was delayed slightly, but the differences were not significant when compared to the controls. However, the hatching percentage of cysts which lacks Hsp70 were reduced following desiccation and freezing. Taken together, these results indicated that Hsp70 possibly plays a role in the stress tolerance but not in the development of diapause-destined embryos of Artemia. This research makes fundamental contributions to our understanding of the role molecular chaperone Hsp70 plays in Artemia, an excellent model organism for diapause studies of the crustaceans.
机译:盐水虾卤虫的雌性通过卵胎生繁殖途径产生自由游泳的无节幼体,或通过卵生途径产生包囊胚胎,称为囊肿,其中生物过程被阻止。虽然先前的研究表明ATP依赖性分子伴侣热休克蛋白70(Hsp70)在保护弗朗西斯卡纳无节幼体免受各种非生物和非生物应激源的侵害方面起着至关重要的作用,但这种蛋白质在滞育胚胎和囊肿发育中的功能仍然未知。本研究采用RNA干扰(RNAi)检测Hsp70在囊肿发展和应激耐受性中的作用,后者通过干燥和冷冻进行,这是卤虫囊肿滞育终止的常用方法。Hsp70 敲低在注射 Hsp70 dsRNA 的女性释放的囊肿中很明显。Hsp70的缺失既不影响囊肿的发育,也不影响囊肿的形态。注射Hsp70 dsRNA的卤虫雌性受精和囊肿释放之间的时间略有延迟,但与对照组相比,差异不显著。然而,缺乏Hsp70的囊肿在干燥和冷冻后孵化率降低。综上所述,这些结果表明,Hsp70可能在应激耐受性中起作用,但在卤虫滞育胚胎的发育中不起作用。这项研究为我们理解分子伴侣Hsp70在卤虫中的作用做出了根本性的贡献,卤虫是甲壳类动物滞育研究的优秀模式生物。

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