Sudhakumari Cheni-Chery, Anitha Arumugam, Murugananthkumar Raju, Tiwari Dinesh Kumar, Bhasker Dharavath, Senthilkumaran Balasubramanian, Dutta-Gupta Aparna
Department of Animal Biology, School of Life Sciences, University of Hyderabad, P.O. Central University, Hyderabad 500046, Telangana, India.
Department of Animal Biology, School of Life Sciences, University of Hyderabad, P.O. Central University, Hyderabad 500046, Telangana, India.
Gen Comp Endocrinol. 2017 Sep 15;251:54-65. doi: 10.1016/j.ygcen.2017.03.006. Epub 2017 Mar 18.
Neuropeptide-Y (NPY) has diverse physiological functions which are extensively studied in vertebrates. However, regulatory role of NPY in relation to brain ontogeny and recrudescence with reference to reproduction is less understood in fish. Present report for the first time evaluated the significance of NPY by transient esiRNA silencing and also analyzed its expression during brain development and gonadal recrudescence in the catfish, Clarias gariepinus. As a first step, full-length cDNA of NPY was cloned from adult catfish brain, which shared high homology with its counterparts from other teleosts upon phylogenetic analysis. Tissue distribution revealed dominant expression of NPY in brain and testis. NPY expression increased during brain development wherein the levels were higher in 100 and 150days post hatch females than the respective age-matched males. Seasonal cycle analysis showed high expression of NPY in brain during pre-spawning phase in comparison with other reproductive phases. Localization studies exhibited the presence of NPY, abundantly, in the regions of preoptic area, hypothalamus and pituitary. Transient silencing of NPY-esiRNA directly into the brain significantly decreased NPY expression in both the male and female brain of catfish which further resulted in significant decrease of transcripts of tryptophan hydroxylase 2, catfish gonadotropin-releasing hormone (cfGnRH), tyrosine hydroxylase and 3β-hydroxysteroid dehydrogenase in brain and luteinizing hormone-β/gonadotropin-II (lh-β/GTH-II) in pituitary exhibiting its influence on gonadal axis. In addition, significant decrease of several ovary-related transcripts was observed in NPY-esiRNA silenced female catfish, indicating the plausible role of NPY in ovary through cfGnRH-GTH axis.
神经肽Y(NPY)具有多种生理功能,在脊椎动物中已得到广泛研究。然而,关于鱼类中NPY在脑发育和与繁殖相关的再生方面的调节作用,人们了解较少。本报告首次通过瞬时esiRNA沉默评估了NPY的重要性,并分析了其在鲶鱼(Clarias gariepinus)脑发育和性腺再生过程中的表达。第一步,从成年鲶鱼脑中克隆出NPY的全长cDNA,系统发育分析表明其与其他硬骨鱼的对应物具有高度同源性。组织分布显示NPY在脑和睾丸中占主导性表达。在脑发育过程中NPY表达增加,其中孵化后100天和150天的雌性鲶鱼中NPY水平高于相应年龄匹配的雄性鲶鱼。季节性周期分析表明,与其他生殖阶段相比,产卵前期脑中NPY表达较高。定位研究显示,视前区、下丘脑和垂体区域大量存在NPY。将NPY-esiRNA直接瞬时沉默到鲶鱼脑中,显著降低了雌雄鲶鱼脑中的NPY表达,进而导致脑中色氨酸羟化酶2、鲶鱼促性腺激素释放激素(cfGnRH)、酪氨酸羟化酶和3β-羟基类固醇脱氢酶以及垂体中促黄体生成素-β/促性腺激素-II(lh-β/GTH-II)的转录本显著减少,表明其对性腺轴有影响。此外,在NPY-esiRNA沉默的雌性鲶鱼中观察到几种与卵巢相关的转录本显著减少,表明NPY可能通过cfGnRH-GTH轴在卵巢中发挥作用。