Wang Linna, Zhu Ying, Xu Wenteng, Shao Changwei, Dong Zhongdian, Li Hailong, Cui Zhongkai, Meng Liang, Guo Hua, Tian Yongsheng, Chen Songlin
Key Lab of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266273, China.
Key Lab of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China.
Biochem Biophys Res Commun. 2017 Dec 16;494(3-4):714-718. doi: 10.1016/j.bbrc.2017.10.126. Epub 2017 Nov 1.
Pod1 encodes a Class II bHLH transcription factor involved in the development of a number of tissues such as gonad, spleen, lungs and heart. However, to date, little is known about its function in teleosts. In this study, we cloned and characterized Pod1 gene from Cynoglossus semilaevis. This gene contains three exons and two introns, with the full-length cDNA of 918 nucleotides that encodes a 183 amino acid protein with a conserved bHLH domain. Realtime quantitative PCR revealed that Pod1 was predominantly expressed in the testes of C. semilaevis. In different stages of testes development, Pod1 expression was undetectable up to 120 days after hatching (dah), and then increased at 210 dah and 1 year after hatching (yah). Furthermore, in situ hybridization (ISH) analysis revealed that Pod1 was mainly localized in the germ cells of testes, but was not detected in ovarian cells; which suggested its possible functions in spermatogenesis of C. semilaevis. The methylation profile analysis of Pod1 genomic sequence in the gonads showed that the differences in their putative promoter regions of Pod1 among ovary, male and pseudo-male testes were not obvious. Thus, further research might be needed to evaluate whether Pod1 expression is regulated by epigenetic level.
Pod1编码一种II类bHLH转录因子，参与性腺、脾脏、肺和心脏等多种组织的发育。然而，迄今为止，关于其在硬骨鱼中的功能知之甚少。在本研究中，我们从半滑舌鳎中克隆并鉴定了Pod1基因。该基因包含三个外显子和两个内含子，全长cDNA为918个核苷酸，编码一个含有保守bHLH结构域的183个氨基酸的蛋白质。实时定量PCR显示，Pod1主要在半滑舌鳎的睾丸中表达。在睾丸发育的不同阶段，孵化后120天（dah）之前未检测到Podl表达，然后在210 dah和孵化后1年（yah）时增加。此外，原位杂交（ISH）分析显示，Pod1主要定位于睾丸的生殖细胞中，但在卵巢细胞中未检测到；这表明其在半滑舌鳎精子发生中可能具有作用。性腺中Pod1基因组序列的甲基化谱分析表明，卵巢、雄性和伪雄性睾丸中Pod1假定启动子区域的差异不明显。因此，可能需要进一步研究来评估Pod1表达是否受表观遗传水平调控。
