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19千道尔顿血淋巴蛋白HP19对米蛾Corcyra cephalonica发育的意义:抗体应用引起的形态学和生化效应

Significance of the 19-kDa hemolymph protein HP19 for the development of the rice moth Corcyra cephalonica: morphological and biochemical effects caused by antibody application.

作者信息

Arif Abul, Gullipalli Damodar, Scheller Klaus, Dutta-Gupta Aparna

机构信息

Department of Animal Sciences, School of Life Sciences, University of Hyderabad, Hyderabad, India.

出版信息

Arch Insect Biochem Physiol. 2007 Sep;66(1):32-44. doi: 10.1002/arch.20195.

Abstract

The hemolymph protein HP19 of the rice moth, Corcyra cephalonica, mediates the 20-hydroxyecdysone (20E)-dependent acid phosphatase (ACP) activity at a nongenomic level. Affinity-purified polyclonal antibody against HP19 (alphaHP19-IgG) was used in the present study to understand the role of HP19 during the postembryonic development of Corcyra. In the in vitro studies, HP19 action was blocked either by immuno-precipitation using alphaHP19-IgG, prior to its addition to the fat body culture or by the addition of the antibody directly to the culture, along with 20E and hemolymph containing HP19. The alphaHP19-IgG blocked the HP19-mediated 20E-dependent ACP activation. In the in vivo studies, the alphaHP19-IgG was injected into the fully developed last (final/Vth) instar larvae of Corcyra, to complex the HP19 in vivo, in order to block the action of HP19. The injection of alphaHP19-IgG resulted in defective development of larvae, which grew either into non-viable larvae or larval-pupal/pupal-adult intermediates relative to the effect of pre-immune IgG injected controls. The present study shows that HP19 plays an important role in controlling the metamorphosis of Corcyra by regulating the 20E-dependent ACP activity. Coupled with the earlier findings, the ecdysteroid hormone regulates this action at a nongenomic level.

摘要

米蛾(Corcyra cephalonica)的血淋巴蛋白HP19在非基因组水平介导20-羟基蜕皮激素(20E)依赖性酸性磷酸酶(ACP)活性。本研究使用针对HP19的亲和纯化多克隆抗体(αHP19-IgG)来了解HP19在米蛾胚后发育过程中的作用。在体外研究中,在将HP19添加到脂肪体培养物之前,通过使用αHP19-IgG进行免疫沉淀,或者将抗体与20E和含有HP19的血淋巴一起直接添加到培养物中,均可阻断HP19的作用。αHP19-IgG阻断了HP19介导的20E依赖性ACP激活。在体内研究中,将αHP19-IgG注射到米蛾发育完全的最后(第五)龄幼虫体内,使体内的HP19形成复合物,以阻断HP19的作用。注射αHP19-IgG导致幼虫发育缺陷,相对于注射免疫前IgG的对照,幼虫要么发育成无法存活的幼虫,要么发育成幼虫-蛹/蛹-成虫中间体。本研究表明,HP19通过调节20E依赖性ACP活性,在控制米蛾变态过程中发挥重要作用。结合早期研究结果,蜕皮甾体激素在非基因组水平调节这一作用。

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