【摘要】 针对兰州鲇人工驯食转化率、苗种培育成活率低等问题，为利用基因编辑技术创制和培育易驯食兰州鲇新种质，开展了兰州鲇神经肽Y基因(NPY)的克隆及其表达分析研究。结果表明：兰州鲇NPY基因cDNA序列全长为775 bp,包括288 bp的开放阅读框，编码95个氨基酸，含有1个跨膜螺旋和1个信号肽胰腺激素类识别结合位点。兰州鲇NPY蛋白序列与南方鲇的一致性最高，为98.95%。荧光定量PCR检测发现NPY mRNA在脑组织中表达量最高，其次是下丘脑，肌肉中表达量最低。不同年龄雄性各组织中NPY mRNA表达水平均高于雌性；且雌雄性别组间均表现出随年龄的增长NPY mRNA表达下降的特征。利用原位杂交技术进行脑组织细胞定位发现，脑细胞中NPY主要在细胞质中表达；不同部位脑组织中主要以前脑的视前区、视顶盖和脑垂体，小脑的豆状核区域以及延脑的尾状核区域表达量较丰富。进一步进行饥饿补偿验证发现，饥饿可引起脑组织NPY表达量显著升高，摄食后NPY表达量显著下降。上述结果表明，NPY基因可能在兰州鲇生长和摄食过程中都具有重要作用，可作为对后续功能基因进行编辑的重要验证并创制新种质，对加快兰州鲇良种新品种选育具有重要的理论意义和技术支撑作用。更多还原

【Abstract】 In view of the low conversion rate of artificial domestication and survival rate of seedling cultivation of Silurus lanzhouensis,in order to use gene editing technology to create and cultivate new germplasm of S.lanzhouensis that was easy to domesticate, the effects of neuropeptide Y gene(NPY) of S.lanzhouensis were studied.The results showed that the full cDNA sequence of the NPY gene was 775 bp in length, with an open reading frame(ORF)of 288 bp, which encoded 95 amino acids and contained a transmembrane helix and a signal peptide binding site for pancreatic hormone recognition.The NPY protein sequence of S.lanzhouensis had the highest consistency with that of Silurus meridionalis,which was 98.95%.The Real-time Fluorescent Quantitative PCR results showed that NPY mRNA expression was the highest in brain tissue, followed by the hypothalamus, and the lowest in muscle.The expression level of NPY mRNA in male tissues of different ages was higher than that in female tissues; and the expression of NPY mRNA decreased with age in both sexes.Localization of brain cells by in situ hybridization found that NPY was mainly expressed in the cytoplasm, and abundant expression was found in the preoptic area, tectum, pituitary, the lentiform nucleus of cerebellum and the caudate nucleus of the medulla oblongata in different parts of the brain.Further verification of starvation compensation showed that the expression of NPY in brain tissue increased significantly after starvation, and decreased significantly after feeding.The above results showed that NPY gene might play an important role in the growth and feeding process of S.lanzhouensis.It could be used as an important verification of the subsequent functional gene editing and the creation of new germplasm, and had important theoretical significance and technical support for accelerating the breeding of new varieties of S.lanzhouensis.更多还原