The kiss2/kiss2r system directly modulates the activity of the pituitary gland in largemouth bass (Micropterus salmoides)

Wuhui Li , Yan Miao , Zexun Zhou , Shi Wang, Fangzhou Hu, Zhongyuan Shen, Lei Zeng, Jie Hu, Ping Wu, RuRong Zhao, Min Tao, Chun Zhang, Qinbo Qin, Shaojun Liu
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Abstract

The kisspeptin system, which involves hypothalamic neuropeptides, plays pivotal roles in the regulation of teleost reproduction. A previous study suggested that the kiss2/kissr2 system in largemouth bass (Micropterus salmoides) was involved in regulating gonadal development through the hypothalamus-pituitary-gonad (HPG) axis. However, whether the kiss2/kiss2r system directly regulates the activity of pituitary gland were not studied. This study aimed to elucidate the detailed mechanism of kisspeptin-induced effects on gonadotropin hormone-producing cells in vivo and in vitro. In situ hybridization analysis revealed very widespread expression of kiss2r mRNA in the optic tectum, hypothalamus and pituitary. In the pituitary, kiss2r-expressing cells were mainly located in the proximal pars distalis and pars intermedia, among which the fshβ- and lhβ-positive cells exhibited expression of kiss2r messengers. Furthermore, primary culture of pituitary cells and peptide administration experiments revealed that the Kiss2-10 peptide stimulates the expression of fshβ and lhβ and the secretion of FSH and LH in 24 h. In addition, histology analysis revealed that three injections of the Kiss2-10 peptide promoted ovary development and increased oocyte size but had little effect on testis development. These results suggested that the intrapituitary kisspeptinergic system, as a hypophysiotropic neuropeptide factor, directly modulates of gonadotroph function. This study will help uncover the reproductive endocrinology network and improve artificial breeding technology for largemouth bass.

kiss2/kiss2r系统直接调节大嘴鲈鱼垂体的活动
涉及下丘脑神经肽的kisspeptin系统在硬骨鱼繁殖的调控中起着关键作用。先前的研究表明,大口黑鲈的kiss2/kissr2系统通过下丘脑-垂体-性腺轴参与性腺发育的调节。然而,关于kiss2/kiss2r系统是否直接调控脑垂体活性的研究尚未见报道。本研究旨在阐明kisspeptin对促性腺激素产生细胞体内外作用的详细机制。原位杂交分析显示kiss2r mRNA在视顶叶、下丘脑和垂体中广泛表达。在垂体中,kiss2r表达细胞主要分布在远部近端和中部,其中fshβ-和lhβ-阳性细胞表达kiss2r信使。此外,垂体细胞原代培养和肽给药实验显示,Kiss2-10肽在24 h内刺激FSH β和LH β的表达以及FSH和LH的分泌。此外,组织学分析显示,三次注射Kiss2-10肽可促进卵巢发育,增加卵母细胞大小,但对睾丸发育影响不大。提示垂体内kisspeptinergic系统作为一种促垂体神经肽因子,直接调节促性腺功能。本研究将有助于揭示大口黑鲈生殖内分泌网络,改进大口黑鲈人工养殖技术。
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