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半变态昆虫美洲大蠊(蜚蠊目,蜚蠊科)超气门基因同源物(RXR/USP)在卵黄发生过程中的分子克隆、特征分析及表达模式

Molecular cloning, characterization, and expression pattern of the ultraspiracle gene homolog (RXR/USP) from the hemimetabolous insect Periplaneta americana (Dictyoptera, Blattidae) during vitellogenesis.

作者信息

Elgendy Azza M, Elmogy Mohamed, Takeda Makio

机构信息

Department of Entomology, Faculty of Science, Cairo University, Giza, Egypt,

出版信息

Mol Biotechnol. 2014 Feb;56(2):126-35. doi: 10.1007/s12033-013-9688-y.

Abstract

Ecdysteroid and sequiterpenoids juvenile hormones play a gonadotrophic role in the insect adult female vitellogenesis. The molecular basis of hormone action has been analyzed in great detail in flies and moths, but rarely in primitive insect orders. The primitive hemimetabolous insect Periplaneta americana was used, as a model, to isolate and characterize, for the first time, two cDNAs of RXR/USP, a component of the heterodimeric ecdysone receptor. These two cDNAs correspond to two isoforms, named PamRXR-S (short form) and PamRXR-L (long form). Both are identical except for 25 amino acids deletion/insertion located in the loop between helices H1 and H3 of the ligand-binding domain. The two isoforms are differentially expressed in different tissues as revealed by RT-PCR and northern blot analysis. In fat body, brain, ovary, and muscle tissues, the predominant form was PamRXR-S, whereas PamRXR-L was abundant in ovaries. The PamRXR transcript was detected during all stages of vitellogenesis in the fat body with different levels. It was little low during the early vitellogenic period (days 2, 3), then a peak of increase was detected during days 4-6 (day 5) which was followed by another peak of increase at the end of vitellogenesis, day 9. We assumed that PamRXR might play a dual role of induction of vitellogenin through JH at early vitellogenesis and suppression through 20E during late vitellogenesis. The present work will pave the way for several other investigations to understand both the ecdysteroid-dependent genetic hierarchy and JH mechanism controlling vitellogenesis in the American cockroach, P. americana.

摘要

蜕皮甾类和倍半萜类保幼激素在昆虫成年雌性卵黄发生过程中发挥促性腺作用。激素作用的分子基础在果蝇和蛾类中已得到详细分析,但在原始昆虫目中却很少见。本研究以原始不完全变态昆虫美洲大蠊为模型,首次分离并鉴定了异源二聚体蜕皮激素受体的一个组分RXR/USP的两个cDNA。这两个cDNA对应两种异构体,分别命名为PamRXR-S(短形式)和PamRXR-L(长形式)。除了位于配体结合域螺旋H1和H3之间环区的25个氨基酸的缺失/插入外,两者完全相同。通过RT-PCR和Northern印迹分析发现,这两种异构体在不同组织中差异表达。在脂肪体、脑、卵巢和肌肉组织中,主要形式是PamRXR-S,而PamRXR-L在卵巢中含量丰富。在脂肪体中,卵黄发生的所有阶段都检测到了PamRXR转录本,但水平不同。在卵黄发生早期(第2、3天)水平较低,然后在第4-6天(第5天)检测到一个增加高峰,随后在卵黄发生末期第9天又出现一个增加高峰。我们推测,PamRXR可能在卵黄发生早期通过保幼激素诱导卵黄原蛋白合成,而在卵黄发生后期通过20E抑制其合成,发挥双重作用。本研究将为其他多项研究铺平道路,以了解美洲大蠊中依赖蜕皮甾类的遗传层次以及控制卵黄发生的保幼激素机制。

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