Collaborative Research: The Molecular Identification and Action of Bursicon, the Insect Cuticle Sclerotizing Hormone

合作研究：昆虫角质层硬化激素 Bursicon 的分子鉴定和作用

• 批准号: 0217471
• 负责人: Karen Mesce
• 金额: $ 2.4万
• 依托单位: University of Minnesota-Twin Cities
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-01 至 2005-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0217471&HistoricalAwards=false
• 关键词: Collaborative; Research; Molecular; Identification; Action

An insect hormone called bursicon is essential for hardening of the new skin (cuticle) after the insect has shed its old one during molting (ecdysis). The timed release and actions of bursicon are critically important for the survival of all insects and, most likely, all arthropods. Although bursicon was discovered in 1935 and is known to be a peptide, its molecular structure has not been established, and remains a major unsolved problem in insect neuroendocrinology. The Honegger lab has purified the hormone and obtained partial amino acid sequences for bursicon from some insects. Recently they were able to use these sequences to identify a particular gene sequence, CG13419, in the genome of the fruitfly Drosophila. The primary aim of this collaborative project is to determine whether the CG13419 gene product is the functional bursicon hormone, as an important step to understanding the actions of bursicon. Molecular techniques for gene expression in eukaryotic cells and overexpression in Drosophila will be combined with the 'ligated fly' bioassay using the related blowfly (Sarcophaga) to test for bursicon activity. If bioactive bursicon is shown, the sequence will be used for cDNA screening in the moth Manduca, a well-studied insect system in which the complex hormonal interactions regulating ecdysis have been illuminated. From the recombinant proteins and synthesized peptides from conserved sequence stretches, new antibodies will be produced to identify neurosecretory cells in the brain, and to clarify the temporal and spatial activation of bursicon release in Manduca. Results from this work will provide a potential breakthrough for understanding how the brain and endocrine systems together orchestrate multiple biochemical events leading to periodic cuticular hardening. Knowledge about this key hormone may provide novel strategies for agriculture in developing insect pest controls, and in managing crustacean aquaculture. In addition, this collaboration between two institutions includes multi-disciplinary postdoctoral training.

昆虫在蜕皮（蜕皮）过程中脱落旧皮肤后，一种称为布尔西康（bursicon）的昆虫激素对于新皮肤（角质层）的硬化至关重要。 bursicon 的定时释放和作用对于所有昆虫（很可能是所有节肢动物）的生存至关重要。 尽管bursicon于1935年被发现并且已知是一种肽，但其分子结构尚未确定，并且仍然是昆虫神经内分泌学中尚未解决的主要问题。 霍内格实验室已经纯化了这种激素，并从一些昆虫中获得了布尔西康的部分氨基酸序列。 最近，他们能够使用这些序列来识别果蝇基因组中的特定基因序列 CG13419。 该合作项目的主要目的是确定 CG13419 基因产物是否是功能性 bursicon 激素，这是了解 bursicon 作用的重要一步。 真核细胞基因表达和果蝇过度表达的分子技术将与使用相关绿头蝇（Sarcophaga）的“结扎蝇”生物测定相结合，以测试 bursicon 活性。 如果显示出具有生物活性的bursicon，则该序列将用于Manduca 蛾中的cDNA 筛选，Manduca 是一种经过充分研究的昆虫系统，其中调节蜕皮的复杂激素相互作用已得到阐明。 从保守序列延伸的重组蛋白和合成肽中，将产生新的抗体来识别大脑中的神经分泌细胞，并阐明曼杜卡中布尔西康释放的时间和空间激活。 这项工作的结果将为理解大脑和内分泌系统如何共同协调导致周期性角质层硬化的多种生化事件提供潜在的突破。 关于这种关键激素的知识可能为农业发展害虫防治和甲壳类水产养殖管理提供新策略。 此外，两个机构之间的合作还包括多学科博士后培训。