National Institute of Nutrition and Seafood Research (NIFES), PO Box 2029 Nordnes, 5817 Bergen, Norway.
Comp Biochem Physiol A Mol Integr Physiol. 2011 Jun;159(2):196-205. doi: 10.1016/j.cbpa.2011.02.026. Epub 2011 Mar 3.
The embryonic stages of Atlantic cod (Gadus morhua) are especially sensitive to incubation temperature. The purpose of the present study was to follow the ontogenetic expression of selected genes of maternal (pou2 and nanog) and zygotic origin (hsp70, hsp90α and stip1), in Atlantic cod embryos under ambient and thermally stressed conditions. The study also investigated how reference genes can be applied to studies on embryonic development, when maternal genes are degraded and the zygotic transcription stabilizes. Three batches of eggs were reared and gene expression profiles from the reference and target genes were determined. The embryos were reared at ambient 6 °C, and 10 °C for continuous long-term and acute short-term heat exposure. Both pou2 and nanog showed reduced expression whereas the zygotic and reference genes showed increased expression until stabilizing at gastrulation, when a normalized ontogenetic expression profile of target genes could be generated. pou2 and nanog were not affected by thermal stress. In contrast, hsp70 and hsp90α were upregulated after short-term heat exposure at the early blastula (hsp70 only), late blastula, 50% epiboly and 90% epiboly stages (hsp90α only). Long-term heat exposure of Atlantic cod embryos upregulated both hsp70 (90% epiboly) and hsp90α (90% epiboly and 20-somites). The results suggest that a cellular defense mechanism is activated even in the earliest stages of embryonic development, a period critical to developmental temperature.
大西洋鳕鱼(Gadus morhua)的胚胎期对孵化温度特别敏感。本研究的目的是在环境和热应激条件下,跟踪母体(pou2 和 nanog)和合子起源(hsp70、hsp90α 和 stip1)的选定基因在大西洋鳕鱼胚胎中的个体发生表达。该研究还调查了在母体基因降解和合子转录稳定时,如何将参考基因应用于胚胎发育研究。培养了三批卵,并确定了参考和靶基因的基因表达谱。将胚胎在环境温度 6°C 和 10°C 下培养,进行持续的长期和急性短期热暴露。pou2 和 nanog 的表达均降低,而合子和参考基因的表达增加,直到原肠胚形成时稳定下来,此时可以生成目标基因的正常个体发生表达谱。pou2 和 nanog 不受热应激的影响。相比之下,hsp70 和 hsp90α 在早期囊胚(仅 hsp70)、晚期囊胚、50%外胚层和 90%外胚层阶段(仅 hsp90α)的短期热暴露后上调。大西洋鳕鱼胚胎的长期热暴露上调了 hsp70(90%外胚层)和 hsp90α(90%外胚层和 20-体节)。结果表明,即使在胚胎发育的最早阶段,即对发育温度至关重要的阶段,也会激活细胞防御机制。