Zheng Yao, Liang Hongwei, Xu Peng, Li Meng, Wang Zaizhao
Shaanxi Key Laboratory of Molecular Biology for Agriculture, College of Animal Science and Technology, Northwest A&F University, 22 Xinong Road, Yangling, 712100, Shaanxi, China.
Fish Physiol Biochem. 2014 Aug;40(4):1141-55. doi: 10.1007/s10695-014-9911-1. Epub 2014 Jan 21.
Dmrt1, an important transcription factor associated with testicular differentiation, is conserved among teleost, which could also be detected in ovaries. In the present study, three isoforms of Pcc-dmrt1s (Pcc-dmrt1a, Pcc-dmrt1b and Pcc-dmrt1c) resulting from alternative splicing of the dmrt1 gene were cloned and characterized in the triploid gynogenetic fish, the Pengze crucian carp. Their mRNA expression profiling was investigated in juvenile developmental stages, tissues of the adult fish, and the juveniles under 84.2 ng/L 17α-methyltestosterone (MT) treatments. Results showed that their putative proteins shared high identities to Dmrt1 in cyprinid fish species. Gene expression profiling in the developmental stages showed that all the three target genes had a highest/lowest expression at 56/40 days post-hatching (dph), respectively. The period of 40 dph appeared to be a key time during the process of the ovary development of Pengze crucian carp. The tissue distribution results indicated that Pcc-dmrt1s were predominantly expressed in hepatopancreas, brain, spleen and ovary of the female fish. MT significantly increased the mRNA expression of Pcc-dmrt1a (all 4-week exposures) and Pcc-dmrt1b (except for week 2), while repressed Pcc-dmrt1c transcripts at all exposure period except for week 2. MT extremely significant repressed cyp19a1a transcripts for 1 week. The present study indicated that MT could influence the ovary development of Pengze crucian carp by disturbing gene expressions of Pcc-dmrt1s and cyp19a1a. Furthermore, the present study will be of great significance to broaden the understanding of masculinizing pathway during ovary development in gynogenetic teleost.
Dmrt1是一种与睾丸分化相关的重要转录因子,在硬骨鱼中保守,在卵巢中也能检测到。在本研究中,从三倍体雌核发育彭泽鲫中克隆并鉴定了dmrt1基因可变剪接产生的三种Pcc-dmrt1亚型(Pcc-dmrt1a、Pcc-dmrt1b和Pcc-dmrt1c)。研究了它们在幼鱼发育阶段、成鱼组织以及84.2 ng/L 17α-甲基睾酮(MT)处理下幼鱼中的mRNA表达谱。结果表明,它们的推定蛋白与鲤科鱼类中的Dmrt1具有高度同源性。发育阶段的基因表达谱显示,所有三个靶基因在孵化后56/40天分别具有最高/最低表达。40日龄似乎是彭泽鲫卵巢发育过程中的关键时期。组织分布结果表明,Pcc-dmrt1在雌鱼的肝胰腺、脑、脾和卵巢中主要表达。MT显著增加了Pcc-dmrt1a(所有4周暴露组)和Pcc-dmrt1b(第2周除外)的mRNA表达,而在除第2周外的所有暴露期均抑制了Pcc-dmrt1c转录本。MT在1周内极显著抑制了cyp19a1a转录本。本研究表明,MT可能通过干扰Pcc-dmrt1和cyp19a1a的基因表达来影响彭泽鲫的卵巢发育。此外,本研究对于拓宽对雌核发育硬骨鱼卵巢发育过程中雄性化途径的理解具有重要意义。