摘要
目的探讨多囊卵巢综合征(PCOS)易感基因dennd1a在斑马鱼模型胚胎发育过程以及成鱼不同器官中的表达,构建适用于PCOS分子机制研究的斑马鱼实验动物模型。方法采用反转录-聚合酶链反应(RT-PCR)及实时荧光定量逆转录聚合酶链反应(qRT-PCR)技术,检测dennd1a基因在斑马鱼中的表达特性。利用胚胎整体原位杂交(WISH)方法,观察dennd1a基因在斑马鱼胚胎内的表达及分布。结果 qRT-PCR结果显示斑马鱼dennd1a mRNA的表达水平随着胚胎的发育而增加,且在多个组织中均有表达,于神经系统和雄性生殖系统中存在高表达。WISH结果表明该基因早期主要在神经系统表达。结论斑马鱼dennd1a基因的表达在胚胎早期发育中发挥重要作用,基因高表达尤其发现于中枢神经系统和雄性生殖系统,提示该基因的表达可能通过影响神经内分泌功能,调控雄激素的合成或释放,诱发PCOS并导致相关表型。
Objective To study the expression characteristics of polycystic ovary syndrome( PCOS) susceptibility gene dennd1a in developing embryos and in adult organs of zebrafish,and to establish zebrafish models for studies of the mechanisms underlying PCOS. Methods Using reverse transcription-polymerase chain reaction( RT-PCR) and realtime fluorescence quantitative reverse transcription polymerase chain reaction( qRT-PCR) techniques,we detected the expression of dennd1a in zebrafish. We also determined the expression pattern of dennd1a gene in zebrafish embryos using whole-mount in situ hybridization( WISH) assays. Results In embryos and juveniles,the expression level of dennd1a increased with age,and in adult animals,the expression was found in different organs. qRT-PCR and WISH showed the expression of dennd1a especially in the nervous system and male reproductive system. Conclusion In early stages of zebrafish development,high expression of dennd1a was found in the nervous system and male reproductive system,indicating this gene may regulate the development and function of the endocrine system and interrupt androgen synthesis and release,thereby inducing PCOS.
作者
刘聪聪
刘洪彬
符江
李雷
LIU Congcong;LIU Hongbin;FU Jiang;LI Lei(Center for Reproductive Medicine,Shandong University,National Research Center for Assisted Reproductive Technology and Reproductive Genetics,Key Laboratory for Reproductive Endocrinology (Shandong University),Ministry of Education,Shandong Provincial Key Laboratory of Reproductive Medicine,Shandong Provincial Clinical Medicine Research Center for Reproductive Health,Jinan 250001,Shandong,China)
出处
《山东大学学报（医学版）》
CAS
北大核心
2019年第1期48-54,共7页
Journal of Shandong University:Health Sciences
基金
国家自然科学基金(81671242)
