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引用本文:肖燃,宋长普,朱阳,黄伟,郑利兵,迟长凤,楼宝.神经肽FF在小黄鱼(Larimichthys polyactis)性腺发育早期的表达与定位分析.海洋与湖沼,2021,52(1):213-221.
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神经肽FF在小黄鱼(Larimichthys polyactis)性腺发育早期的表达与定位分析
肖燃1, 宋长普1, 朱阳1, 黄伟1, 郑利兵1, 迟长凤1, 楼宝2
1.浙江海洋大学海洋科学与技术学院 国家海洋设施养殖工程技术研究中心 海洋生物种质发掘与利用国家地方联合实验室 舟山 316022;2.浙江省农业科学院水生生物研究所 杭州 310021
摘要:
神经肽FF是属于RFamide相关肽家族中的一种八肽,在多种生物过程中具有调节功能。本文开展了小黄鱼(Larimichthys polyactis)神经肽FF相关基因(LpNPFF:Accession No.MT012894)的组织表达与定位分析研究。实时荧光定量PCR结果显示,小黄鱼神经肽FF基因在性未成熟期(第Ⅱ期)的小黄鱼的脑和性腺中表达较高,其次是肌肉和脾脏,在胃和肠中表达相对较少;原位杂交结果显示,在小黄鱼中脑的视顶盖(OTec)、圆环体(TL)、结节前核(NAT)、外侧结节核(NLT)、下丘脑弥散核(NDLI)以及延脑(MO)中检测到大量LpNPFF mRNA阳性的细胞,而端脑和小脑部分的表达较少。同时,在小黄鱼第Ⅱ期卵巢细胞的细胞质和核仁中也能观察到明显的阳性信号;免疫组织化学结果显示,在小黄鱼脑部表层细胞和卵巢壁周围的结缔组织中可见明显的RFamide免疫阳性纤维。上述研究结果有助于神经肽进化的深入研究,也为进一步探索NPFF相关基因在生殖调控方面的潜在功能提供了新的思路。
关键词:  小黄鱼(Larimichthys polyactis)  神经肽FF (neuropeptide FF)  组织表达与定位
DOI:10.11693/hyhz20200500146
分类号:Q789;S965.3
基金项目:国家自然科学基金项目,31872547号;浙江省重点科研专项,2017C02013-2号;浙江省自然科学基金项目,LY20C190007号。
EXPRESSION AND LOCALIZATION OF THE NEUROPEPTIDE FF GENE FROM LARIMICHTHYS POLYACTIS DURING PRIMITIVE GONADAL PERIOD
XIAO Ran1, SONG Chang-Pu1, ZHU Yang1, HUANG Wei1, ZHENG Li-Bing1, CHI Chang-Feng1, LOU Bao2
1.National and Provincial Joint Laboratory of Exploration and Utilization of Marine Aquatic Genetic Resources, National Engineering Research Center of Marine Facilities Aquaculture, School of Marine Science and Technology, Zhejiang Ocean University, Zhoushan 316022, China;2.Institute of Hydrobiology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
Abstract:
Neuropeptide FF is an octapeptide belonging to the RFamide-related peptide family, and has regulatory functions in a variety of biological processes. We analyzed the expression and positioning of NPFF related gene (LpNPFF:Accession No.MT012894) from Larimichthys polyactis. The results of qRT-PCR analysis show that LpNPFF was expressed mainly in the brain and gonads of L. polyactis (stage Ⅱ), followed by muscles and spleen, and relatively less expressed in the stomach and intestinal tissues. The localization and distribution of LpNPFF in L. polyactis were revealed by in situ hybridization and immunohistochemistry. In the in situ hybridization, a large number of LpNPFF mRNA positive cells were detected in optic tectum (OTec), torus lobe (TL), nodules pronucleus (NAT), lateral nodules (NLT), the hypothalamus (NDLI) of the midbrain, and medulla oblongata (MO), while less positive cells were detected in telencephalon and cerebellum in L. polyactis. And obvious positive signals were also observed in the cytoplasm and nucleolus in L. polyactis ovary cells at stage Ⅱ. In addition, immunohistochemical results show that RFamid immunopositive signals were visible mainly in the surface cells of the brain and the connective tissue around the ovarian wall of L. polyactis. These results contribute to future studies of neuropeptide evolution or also provide new ideas for further exploration of the reproductive regulation of NPFF-related genes.
Key words:  Larimichthys polyactis  neuropeptide FF  tissue expression and localization
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