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蜕皮抑制因子 Krüppel 同源物 1 的激素调控及其在桑蚕中的发育作用。

Hormonal regulation and developmental role of Krüppel homolog 1, a repressor of metamorphosis, in the silkworm Bombyx mori.

机构信息

National Institute of Agrobiological Sciences, Ohwashi 1-2, Tsukuba, Ibaraki 305-8634, Japan.

Faculty of Agriculture and Life Sciences, Hirosaki University, Hirosaki 036-8561, Japan.

出版信息

Dev Biol. 2014 Apr 1;388(1):48-56. doi: 10.1016/j.ydbio.2014.01.022. Epub 2014 Feb 4.

Abstract

Juvenile hormone (JH) has an ability to repress the precocious metamorphosis of insects during their larval development. Krüppel homolog 1 (Kr-h1) is an early JH-inducible gene that mediates this action of JH; however, the fine hormonal regulation of Kr-h1 and the molecular mechanism underlying its antimetamorphic effect are little understood. In this study, we attempted to elucidate the hormonal regulation and developmental role of Kr-h1. We found that the expression of Kr-h1 in the epidermis of penultimate-instar larvae of the silkworm Bombyx mori was induced by JH secreted by the corpora allata (CA), whereas the CA were not involved in the transient induction of Kr-h1 at the prepupal stage. Tissue culture experiments suggested that the transient peak of Kr-h1 at the prepupal stage is likely to be induced cooperatively by JH derived from gland(s) other than the CA and the prepupal surge of ecdysteroid, although involvement of unknown factor(s) could not be ruled out. To elucidate the developmental role of Kr-h1, we generated transgenic silkworms overexpressing Kr-h1. The transgenic silkworms grew normally until the spinning stage, but their development was arrested at the prepupal stage. The transgenic silkworms from which the CA were removed in the penultimate instar did not undergo precocious pupation or larval-larval molt but fell into prepupal arrest. This result demonstrated that Kr-h1 is indeed involved in the repression of metamorphosis but that Kr-h1 alone is incapable of implementing normal larval molt. Moreover, the expression profiles and hormonal responses of early ecdysone-inducible genes (E74, E75, and Broad) in transgenic silkworms suggested that Kr-h1 is not involved in the JH-dependent modulation of these genes, which is associated with the control of metamorphosis.

摘要

保幼激素(JH)具有抑制昆虫幼虫期过早变态的能力。Krüppel 同源物 1(Kr-h1)是一种早期 JH 诱导基因,介导 JH 的这种作用;然而,Kr-h1 的精细激素调节及其抗变态作用的分子机制尚不清楚。在这项研究中,我们试图阐明 Kr-h1 的激素调节和发育作用。我们发现,家蚕五龄幼虫表皮中的 Kr-h1 表达是由咽侧体(CA)分泌的 JH 诱导的,而 CA 不参与预蛹期 Kr-h1 的短暂诱导。组织培养实验表明,预蛹期 Kr-h1 的短暂峰值可能是由来自 CA 和蜕皮激素的预蛹激增的腺体以外的 JH 共同诱导的,尽管不能排除未知因素(s)的参与。为了阐明 Kr-h1 的发育作用,我们生成了过表达 Kr-h1 的转基因家蚕。转基因家蚕正常生长直至吐丝阶段,但它们的发育在预蛹期被阻止。在家蚕五龄幼虫期去除 CA 的转基因家蚕没有经历过早化蛹或幼虫-幼虫蜕皮,但进入预蛹期停滞。这个结果表明,Kr-h1 确实参与了变态的抑制,但 Kr-h1 本身不能实施正常的幼虫蜕皮。此外,转基因家蚕中早期蜕皮激素诱导基因(E74、E75 和 Broad)的表达谱和激素反应表明,Kr-h1 不参与 JH 依赖的这些基因的调节,这与变态的控制有关。

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