摘要
为探讨胰岛素样生长因子-Ⅰ(Insulin like growth factor-Ⅰ,IGF-Ⅰ)对宽鳍鱲(Zacco platypus)繁殖期前后生长性状的影响,开展了宽鳍鱲IGF-Ⅰ基因序列的克隆及表达定位分析。宽鳍鱲IGF-Ⅰ基因全长13707 bp,包含5个外显子、4个内含子,其中5个外显子长度分别为222、160、182、36和829 bp;内含子长度分别为1194、7771、254和1879 bp,推测开放阅读框为486 bp,编码161个氨基酸。实时荧光定量PCR(Real-time qPCR,RT-qPCR)结果显示,宽鳍鱲IGF-ⅠmRNA在肝脏组织中的表达水平最高,其次是性腺、脾脏、心脏和脑,在肾脏中表达水平最低。在7—8月,处于繁殖期的性成熟宽鳍鱲性腺中IGF-Ⅰ表达水平显著上升,繁殖期过后回落至最低值。在其他组织中IGF-Ⅰ表达水平在生长发育过程和繁殖期前后波动不大,且雄鱼大多数组织中IGF-Ⅰ基因平均表达水平高于雌鱼。荧光原位杂交技术(Fluorescence in situ hybridization,FISH)定位显示,宽鳍鱲IGF-Ⅰ基因基本为胞浆阳性,少数为核阳性,在肝组织中呈全胞质性分布,在性腺组织的精母细胞、卵泡膜及卵泡液中阳性表达。上述结果表明宽鳍鱲IGF-Ⅰ基因表达模式具有性别差异性,推测精巢中IGF-Ⅰ在繁殖期的高表达是宽鳍鱲雄性成体大于雌性成体的原因之一。研究结果为宽鳍鱲的性二态和人工繁育的研究提供参考资料。
In order to investigate the effect of insulin-like growth factor-Ⅰ(IGF-Ⅰ)on the growth traits of Zacco platypus,IGF-Ⅰgene sequence was cloned and analyzed.The gene was 13707 bp in length and contained five exons and four introns.The five exons of IGF-Ⅰwere 222,160,182,36,and 829 bp long,and the four introns were 1194,7771,254,and 1879 bp long,respectively.The open reading frame was estimated to be 486 bp,encoding 161 amino acids.The results of real-time qPCR(RT-qPCR)showed that the IGF-ⅠmRNA in liver tissue was the highest,followed by gonad,spleen,heart and brain,and the lowest expression in kidney.From July to August,IGF-Ⅰin the sexually mature Zacco platypus gonads increased significantly during the breeding period and fell to the lowest value after the breeding period.The expression level of IGF-Ⅰin other tissues did not fluctuate greatly during the growth and development process and before and after reproduction.The average expression level of IGF-Ⅰgene in most tissues of male individuals were higher than that of females.Fluorescence in situ hybridization(FISH)mapping showed that IGF-Ⅰgene was broadly cytoplasmic-positive and a few were nuclear-positive,showing a full cytoplasmic distribution in liver tissues.It also positively expressed in spermatocytes,follicular membranes and follicular fluid.The above results indicated that the IGF-Ⅰgene expression pattern of Zacco platypus was gender-specific.It is speculated that the high expression of IGF-Ⅰin the testis during reproduction is one of the reasons why males are larger than females.The results of this study provide reference materials for the study of sexual dimorphism and artificial breeding of Zacco platypus.
作者
沈铭浩
朱迦玥
吴涛
任天惠
张哲盼
熊黎
刘芳玲
郑善坚
SHEN Ming-Hao;ZHU Jia-Yue;WU Tao;REN Tian-Hui;ZHANG Zhe-Pan;XIONG Li;LIU Fang-Ling;ZHENG Shan-Jian(College of Chemistry and Life Sciences,Zhejiang Normal University,Jinhua 321004,China;Key Laboratory of Wildlife Biotechnology and Conservation and Utilization of Zhejiang Province,Zhejiang Normal University,Jinhua 321004,China)
出处
《水生生物学报》
CAS
CSCD
北大核心
2020年第6期1182-1190,共9页
Acta Hydrobiologica Sinica
基金
浙江省重点研发计划项目(2019C02049和2020C02014)资助。
关键词
宽鳍鱲
胰岛素样生长因子-Ⅰ
克隆
序列分析
表达模式
Zacco platypus
Insulin-like growth factor-Ⅰgene
Clone
Sequence analysis
Expression analysis
