拟穴青蟹短神经肽F（sNPF）生殖调控作用及机制

By far, the studies of neuropeptides in crustaceans have been focused on crustacean hyperglycemic hormone (CHH) superfamily in the eyestalk ganglia, while the structure and functions of small neuropeptides are poorly understood. Since short neuropeptide F(sNPF) has some potential application advantages, this project aims to study its reproductive role and mechanism in the mud crab Scylla paramamosain. The main research contents are as following: (1) Study the sequence, tissue distribution, cellular localization and ligand-receptor affinity interaction of sNPF and its receptor (sNPFR), in order to understand their characteristics and relationship; (2) Study the regulatory function of sNPF over the nervous organ, endocrine organ and ovary, in order to elucidate the reproductive regulation pathway of sNPF. In this project, experimental techniques such as cell transfection, calcium imaging, patch clamp, RNA interference and etc,.will be employed to clarify the reproductive regulation mechanism of sNPF in S. paramamosain from the molecular, cellular, organic and individual levels. It is for the first time that a comprehensive research on the reproductive regulation pathway of crustacean sNPF will be carried out in this project. Meanwhile, it is for the first time that we propose that sNPF could regulate reproduction as paracrine factors in ovary in S. paramamosain. In all, this research will provide scientific supports for crustacean breeding technique, and have both theoretical and practical significance.

目前甲壳动物神经肽的研究主要集中在眼柄神经节高血糖激素（CHHs）超家族上，有关小分子神经肽结构与功能的了解甚少。本项目针对小分子神经肽的潜在应用优势，对拟穴青蟹的一种小分子神经肽——短神经肽F（sNPF）的生殖调控作用及机制开展研究，内容主要包括：（1）研究sNPF及受体的基因序列、组织分布、细胞定位以及配体-受体亲和作用，了解二者的特性与关系；（2）研究sNPF对神经器官、内分泌器官和卵巢的调控作用，查明sNPF的生殖调控途径。本项目拟应用细胞转染、钙离子成像、膜片钳、RNA干扰等先进的实验技术，从分子、细胞、器官和个体水平，深入阐明sNPF在拟穴青蟹中的生殖调控机制。本项目首次对甲壳动物sNPF的生殖调控途径开展全面综合的探索研究，也是首次提出sNPF可能作为卵巢内的旁分泌因子调控生殖的观点。该研究成果将为甲壳动物繁殖技术提供科学依据，具有重要的理论意义和潜在的应用价值。

长期以来，甲壳动物神经肽的研究主要关注眼柄CHH家族，而对小分子神经肽的结构与功能缺少了解。本项目针对小分子神经肽的潜在应用优势，对拟穴青蟹的一种小分子神经肽——短神经肽F（sNPF）的内分泌作用及机制开展研究，主要成果如下：（1）查明了拟穴青蟹sNPF及受体的基因序列、组织分布、细胞定位以及配体-受体亲和作用；（2）确定了拟穴青蟹sNPF可以作为神经激素和自分泌/旁分泌因子调控卵巢发育。（3）发现sNPF对拟穴青蟹肝胰腺和血细胞的免疫功能具有促进作用。本项目首次对甲壳动物sNPF的生殖调控途径开展全面综合的探索研究，并提出sNPF可能作为卵巢内的旁分泌因子调控生殖的观点。此外，证实了甲壳动物存在神经内分泌-免疫调节机制。本研究成果将为甲壳动物繁殖技术和病害防治提供科学依据，具有重要的理论意义和潜在的应用价值。