Molecular Analysis of Eclosion Hormone in the Tobacco Hornworm

这个项目将研究分子结构和 调节神经肽、羽化激素的合成 (EH)烟草天蛾Manduca sexta. EH的合成 在大脑和腹侧中枢神经系统中， 蜕皮以引发预先设定的神经活动模式 最终蜕皮。 62氨基酸肽最近 已经测序，我们打算分离EH基因， 基于蛋白质设计的合成寡核苷酸探针 顺序 EH基因的分离将提供 研究神经肽的合成和作用的机会 在昆虫相对简单的神经系统中 可识别的神经元 我们将通过以下方法来研究EH基因的结构： 核酸测序和转录图谱研究， 鉴定来自激素原的其他潜在肽。 我们将确定EH mRNA的发育调控 合成，并将分析调节内分泌因子， 它的表达。 将确定EH转录物的细胞定位 通过在大脑和细胞中的原位杂交 与腹侧CNS相关，EH的两个来源。 最后， 其他昆虫中EH样序列的存在将是 通过与EH基因的杂交进行检测。

This project will examine the molecular structure and the regulation of synthesis of the neuropeptide, eclosion hormone (EH), in the tobacco hornworm, Manduca sexta. EH is synthesized both in the brain and in the ventral CNS and released before each molt to elicit a preprogrammed pattern of neural activity culminating in ecdysis. The 62 amino acid peptide has recently been sequenced and we intend to isolate the EH gene using synthetic oligonucleotide probes designed based on the protein sequence. The isolation of the EH gene will offer the opportunity to study the synthesis and action of a neuropeptide in the relatively simple nervous system of an insect with it's identifiable neurons. We will examine the structure of the EH gene by a combination of nucleic acid sequencing and transcript mapping studies and will identify other potential peptides derived from the prohormone. We will determine the developmental regulation of EH mRNA synthesis and will analyze the endocrine factors that regulate its expression. The cellular localization of the EH transcript will be determined by in situ hybridization both in the brain and in the cells associated with the ventral CNS, the two sources of EH. Finally, the presence of EH-like sequences in other insects will be examined by hybridization to the EH gene.