RUI: Identifying reproductive roles for the Super-conserved Receptors Expressed in Brain (SREB) G protein-coupled receptor family using novel agonists and a comparative fish model

SREBs(在脑中表达的超保守受体)是脊椎动物体内的一组激素受体蛋白，其功能尚不清楚。这种知识差距源于缺乏与细胞表面SREB结合的已证实的激素。以前的研究已经支持SREB在大脑、肠道和性腺中的功能作用，这些作用可能在动物身上保存下来。这个项目应用了新的人工合成的分子，这些分子已知与三种鱼类卵巢中的SREB受体结合，以评估SREB在繁殖中的功能。这些物种是根据先前发现的遗传差异选择的，导致这些物种的SREB亚型分布不同。结果将在不同种类的鱼之间进行比较，以确定独特和共享的功能。因此，对SREB功能的进一步了解可能为未来在动物繁殖和水产养殖中的商业应用奠定基础。这项研究将生成数据库，以培养缅因州西部农村地区的本科生和高中生在生物信息学方面的研究经验，缅因州西部农村地区第一代大学生的比例很高。将培训当地高中教师在课堂上使用这些数据库，并开发可持续使用的独立生物信息学模块。这些教育活动是一个可扩展的模式，将生物信息学培训带给服务不足的学生群体，为生物技术劳动力的发展做出贡献。此外，通过与佛罗里达大学的两名研究人员合作，该项目将有助于培养一名研究生和博士后研究员。最近针对孤儿G蛋白偶联受体家族SREB的新型和特定激动剂的开发使理解受体功能的生理机制成为可能。两个合成和验证的激动剂分子正与潜在的内源性配体(菲尼辛)结合用于体外卵巢治疗和三种转基因鱼类的RNA测序：斑马鱼(Danio Rerio)、乳鼠(Fundulus Heterocltes)和河豚(Dichotomyctere Nigroviridis)。这些物种中的每一个都表现出不同的SREB受体互补，这使得比较转录转录方法能够识别整个家族中保守的生殖功能以及SREB成员特有的效应。这些方法将与类固醇定量和RNAScope结合使用，分别识别与SREB基因表达相关的生殖激素变化和特定性腺细胞类型。这种比较的多物种模型的发展影响了未来SREB的研究，而不仅仅是生殖，例如对大脑和肠道的研究，在那里SREB也涉及保守的功能。该项目由生物-IOS-生理机制和生物力学计划和既定的激励竞争性研究计划(EPSCoR)共同资助。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

SREBs (Super-conserved Receptors Expressed in Brain) are a group of hormone receptor proteins in vertebrate animals whose function is poorly understood. This knowledge gap stems from a lack of verified hormones that bind to SREBs on the surface of cells. Previous studies have supported functional roles for SREBs in the brain, gut, and gonads that may be conserved in animals. This project applies novel, artificially synthesized molecules that are known to bind to SREB receptors in ovaries of three fish species to assess SREB function in reproduction. The species were chosen based on previously identified genetic differences resulting in different profiles of SREB subtypes across these species. Results will be compared among the fish species to identify unique and shared functions. The resulting improved understanding of SREB functions may provide a foundation for future commercial applications in animal reproduction and in aquaculture. The research will generate databases that will foster undergraduate and high-school student research experiences in bioinformatics in rural western Maine, which has a high proportion of first-generation college students. Local high-school teachers will be trained to use these databases in their classes and to develop independent bioinformatics modules for sustainable use. These educational activities serve as a scalable model to bring bioinformatics training to under-served student populations, contributing to biotechnology workforce development. In addition, through a collaboration with two researchers at the University of Florida, the project will contribute to training of a graduate student and post-doctoral fellow.The recent development of novel and specific agonists for the orphan G protein-coupled receptor family SREB has enabled new approaches to understand the physiological mechanisms of receptor function. Two synthesized and validated agonist molecules are being used in combination with a potential endogenous ligand (phoenixin) in in vitro ovary treatments and RNA-sequencing of three genetically enabled fish species: zebrafish (Danio rerio), mummichog (Fundulus heteroclitus), and pufferfish (Dichotomyctere nigroviridis). Each of these species exhibits a different complement of SREB receptors, which enables comparative transcriptomic approaches to identify both conserved reproductive functions across the family as well as SREB member-specific effects. These methods will be used in combination with steroid quantifications and RNAScope to identify reproductive hormone changes and specific gonadal cell types associated with SREB gene expression, respectively. The development of this comparative multispecies model impacts future SREB studies beyond reproduction, such as investigations in the brain and gut where SREBs have also been implicated in conserved functions. This project is jointly funded by the BIO-IOS-Physiological Mechanisms and Biomechanics Program and the Established Program to Stimulate Competitive Research (EPSCoR).This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.