State Key Laboratory of Biocontrol, Institute of Aquatic Economic Animals, Sun Yat-Sen University, Guangzhou 510275, China.
Gen Comp Endocrinol. 2012 Oct 1;179(1):99-106. doi: 10.1016/j.ygcen.2012.07.018. Epub 2012 Aug 8.
In the present study, the first full-length cDNA encoding Neuropeptide Y (NPY) was cloned from the brain of Japanese eel (Anguilla japonica). The open reading frame of Japanese eel NPY gene is 294 bp in length, encoding a precursor protein of 97 amino acids, which contains a 36-amino-acid mature peptide. Sequence analysis showed that the Japanese eel NPY peptide is similar to that of other species. Real-time PCR revealed that NPY in Japanese eel is mainly expressed in the brain, especially in the hypothalamus and the optic tectum thalamus. The effect of a negative energy balance on NPY gene expression was examined subsequently. The mRNA level of NPY in the hypothalamus and the optic tectum thalamus showed a pronounced increase after 4 days of food deprivation. The biological activities of Japanese eel NPY were further investigated in vivo and in vitro. Intraperitoneal injection of the NPY peptide into Japanese eel could potently elevate the expression of the mammalian gonadotropin-releasing hormone (mGnRH) in hypothalamus and the follicle-stimulating hormone beta (FSHβ), the luteinizing hormone beta (LHβ) and growth hormone (GH) in pituitary. In static incubation studies, the stimulatory effects of NPY on mGnRH expression in hypothalamic fragments and on FSHβ, LHβ and GH expression in pituitary cells were also observed. However, in vivo and in vitro studies showed that NPY exhibits an inhibitory action on the expression of thyroid-stimulating hormone beta (TSHβ) in pituitary. The results indicate that NPY is involved in the regulation of multiple physiological processes in Japanese eel.
在本研究中,首次从日本鳗鲡(Anguilla japonica)脑中克隆出编码神经肽 Y(NPY)的全长 cDNA。日本鳗鲡 NPY 基因的开放阅读框长 294bp,编码 97 个氨基酸的前体蛋白,其中包含 36 个氨基酸的成熟肽。序列分析表明,日本鳗鲡 NPY 肽与其他物种相似。实时 PCR 显示,NPY 在日本鳗鲡中主要表达于脑,特别是下丘脑和视顶盖丘脑。随后研究了负能平衡对 NPY 基因表达的影响。禁食 4 天后,下丘脑和视顶盖丘脑的 NPY mRNA 水平显著增加。进一步在体内和体外研究了日本鳗鲡 NPY 的生物学活性。腹腔内注射 NPY 肽可强烈上调下丘脑哺乳动物促性腺激素释放激素(mGnRH)和垂体卵泡刺激素β(FSHβ)、黄体生成素β(LHβ)和生长激素(GH)的表达。在静态孵育研究中,还观察到 NPY 对下丘脑片段中 mGnRH 表达以及对垂体细胞中 FSHβ、LHβ 和 GH 表达的刺激作用。然而,体内和体外研究表明,NPY 对垂体中促甲状腺素β(TSHβ)的表达具有抑制作用。结果表明,NPY 参与了日本鳗鲡多种生理过程的调节。