PHYSIOLOGY OF A MOSQUITO HOMOLOG OF ALLATOTROPIN

促尿素的蚊子同系物的生理学

• 批准号: 2068430
• 负责人: JAN A VEENSTRA
• 金额: $ 8.44万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-12-01 至 1997-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2068430
• 关键词: Aedes; antiserum; electron microscopy; enzyme linked immunosorbent assay; guinea pigs; hemolymph; high performance liquid chromatography; hormone biosynthesis; hormone regulation /control mechanism; immunoelectron microscopy; immunologic assay /test; juvenile hormones; laboratory rabbit; neuropeptide receptor; neuropeptides; ovary; peptide structure; protein structure function; reproduction

Juvenile hormone plays an important role in the reproduction of the mosquito Aedes aegypti, and the rate of its synthesis by the corpora allata is well regulated. In other insect species this regulation is accomplished by neuropeptides. One of them is a recently characterized allatotropin from Manduca sexta, which stimulates juvenile hormone synthesis in adult, but not larval corpus allatum. A peptide homologous to this allatotropin has been isolated from a locust as a stimulator of oviduct contractions. A peptide homologous to this allatotropin has been isolated from a locust as a stimulator of oviduct contractions. This suggests that members of this novel peptide family may play multiple roles in insect reproduction. I have raised antisera against Manduca allatotropin and used them to study its distribution in Manduca and Aedes by HPLC and ELISA. In Aedes a peptide was found that is very similar to, but different from the allatotropin from Manduca. Using immunocytology, this material was detected both in brain neurons with axon terminals in the corpus allatum, as well as in neurosecretory cells in the abdominal ganglia and neurohemal organs associated with these ganglia. Its presence in homologous structures in Aedes and Manduca, as well as its similarity in physico-chemical characteristics to Manduca allatotropin, suggests that this peptide is a homologue of Manduca allatotropin. This is therefore a putative mosquito allatotropin. The specific aims of this research proposal are: (1) To isolate, chemically characterize, and synthetize this putative Aedes allatotropin. (2) To determine its effects on juvenile hormone biosynthesis and oviduct contractions in the mosquito, and the inhibitory effects of antibodies against this peptide on reproduction. (3) To raise specific antisera to the synthetic peptide and use those to study its distribution within the mosquito and identify putative neural and neurohormonal release sites, and to show that it is released from these sites using immunoelectron microscopy. (4) To quantify concentrations of this putative Aedes allatotropin in the hemolymph in relation to the reproductive cycle with a sensitive sandwich immuno assay. Disruption of the interactions of insect neuropeptides with their receptors has been suggested as a target for a novel generation of insecticides. However, until the physiological relevance of these neuropeptides has been established it is very difficult to select any of them for detailed research in this direction. Manipulation of the putative mosquito allatotropin could have deleterious effects on mosquito reproduction. The work proposed here will yield data as to the physiological relevance of this neuropeptide.

青少年激素在繁殖中起重要作用 蚊子埃及埃及及其合成率 Allata受到良好的监管。 在其他昆虫物种中，这个法规是 通过神经肽完成。 其中之一是最近的特征 甘杜卡·塞克斯塔（Manduca Sexta 成年人的合成，但不是幼虫属的菜单。 肽同源 对此，腺激素已从蝗虫中分离为刺激剂 输卵管收缩。 与这种腺体性蛋白质同源的肽已经 从蝗虫分离为输卵管收缩的刺激器。 这 暗示这个新颖的肽家族的成员可能会玩多个 在昆虫繁殖中的作用。 我已经针对甘达阿拉托蛋白提出了抗血清，并用它们 研究了HPLC和ELISA在Manduca和Aedes中的分布。 在艾德斯 发现肽与 来自甘达的Allatotropin。 使用免疫细胞学，该材料是 在Allatum语料库中的轴突终端中检测到两者， 以及在腹部神经节中的神经分泌细胞中 与这些神经节相关的神经性器官。 它的存在 艾德斯和曼达的同源结构，及其相似之处 甘达藻蛋白的物理化学特征，表明 该肽是甘达藻蛋白的同源物。 因此，这是 推定的蚊子藻蛋白。 该研究建议的具体目的是： （1）分离，化学表征并合成该推定的 艾德斯·阿拉托罗蛋白。 （2）确定其对青少年激素生物合成和 蚊子中的卵形收缩和抑制作用 对该肽的抗体在繁殖时进行抗体。 （3）提出特定的抗血清对合成肽并使用 研究其在蚊子中的分布并确定推定的 神经和神经激素释放位点，并表明它已释放 使用免疫电子显微镜从这些位点进行。 （4）量化该假定伊蚊的浓度allatotropin 与生殖循环有关的血淋巴 三明治免疫测定法。 昆虫神经肽与它们的相互作用的破坏 已建议受体作为新产生的靶标 杀虫剂。 但是，直到这些的生理相关性 已经建立了神经肽，很难选择任何一个 他们朝着这个方向进行详细研究。 操纵 假定的蚊子藻蛋白可能会对蚊子产生有害影响 生殖。 这里提出的工作将产生有关 该神经肽的生理相关性。