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卤虫胚胎中滞育特异性分子伴侣的合成由热休克因子 1(Hsf1)的数量和位置决定。

The synthesis of diapause-specific molecular chaperones in embryos of Artemia franciscana is determined by the quantity and location of heat shock factor 1 (Hsf1).

机构信息

Department of Biology, Dalhousie University, Halifax, NS, B3H 4R2, Canada.

出版信息

Cell Stress Chaperones. 2019 Mar;24(2):385-392. doi: 10.1007/s12192-019-00971-7. Epub 2019 Jan 30.

Abstract

The crustacean, Artemia franciscana, displays a complex life history in which embryos either arrest development and undertake diapause as cysts or they develop into swimming nauplii. Diapause entry is preceded during embryogenesis by the synthesis of specific molecular chaperones, namely the small heat shock proteins p26, ArHsp21, and ArHsp22, and the ferritin homolog, artemin. Maximal synthesis of diapause-specific molecular chaperones is dependent on the transcription factor, heat shock factor 1 (Hsf1), found in similar amounts in cysts and nauplii newly released from females. This investigation was performed to determine why, if cysts and nauplii contain comparable amounts of Hsf1, only cyst-destined embryos synthesize diapause-specific molecular chaperones. Quantification by qPCR and immunoprobing of Western blots, respectively, demonstrated that hsf1 mRNA and Hsf1 peaked by day 2 post-fertilization in embryos that were developing into cysts and then declined. hsf1 mRNA and Hsf1 were present in nauplii-destined embryos on day 2 post-fertilization, but in much smaller amounts than in cyst-destined embryos, and they increased in quantity until release of nauplii from females. Immunofluorescent staining revealed that the amount of Hsf1 in nuclei was greatest on day 4 post-fertilization in cyst-destined embryos but could not be detected in nuclei of nauplius-destined embryos at this time. The differences in quantity and location of Hsf1 explain why embryos fated to become cysts and eventually enter diapause synthesize p26, ArHsp21, ArHsp22, and artemin, whereas nauplius-destined embryos do not produce these molecular chaperones.

摘要

卤虫,弗朗西斯科卤虫,在其复杂的生活史中表现出胚胎要么停滞发育并形成包囊进入休眠,要么发育成游动的无节幼体。在胚胎发生过程中,休眠的进入先于特定分子伴侣的合成,即小热休克蛋白 p26、ArHsp21 和 ArHsp22 以及铁蛋白同源物 artemin。休眠特异性分子伴侣的最大合成依赖于在刚从母体中释放的包囊和无节幼体中含量相似的转录因子热休克因子 1(Hsf1)。进行这项研究是为了确定为什么如果包囊和无节幼体含有相当数量的 Hsf1,只有注定要形成包囊的胚胎才会合成休眠特异性分子伴侣。通过 qPCR 定量和 Western 印迹免疫探测分别证明 hsf1 mRNA 和 Hsf1 在发育成包囊的胚胎中于受精后第 2 天达到峰值,然后下降。受精后第 2 天,无节幼体定向胚胎中存在 hsf1 mRNA 和 Hsf1,但数量比包囊定向胚胎少得多,并且在从母体中释放无节幼体之前,其数量增加。免疫荧光染色显示,受精后第 4 天,包囊定向胚胎中 Hsf1 的核内含量最大,但此时无法检测到无节幼体定向胚胎的核内 Hsf1。Hsf1 的数量和位置的差异解释了为什么注定要形成包囊并最终进入休眠的胚胎合成 p26、ArHsp21、ArHsp22 和 artemin,而无节幼体定向胚胎不产生这些分子伴侣。

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