Henan University of Science and Technology, Luoyang, 471000, China; Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Science (CAFS), Laboratory for Marine Fisheries Science and Food Production Processes, Pilot National Laboratory for Marine Science and Technology Qingdao, 266071, China.
Guangdong South China Sea Key Laboratory of Aquaculture for Aquatic Economic Animals, Guangdong Ocean University, Zhanjiang, 524088, China.
Gene Expr Patterns. 2021 Mar;39:119163. doi: 10.1016/j.gep.2020.119163. Epub 2020 Dec 28.
Desert hedgehog (dhh) is a gene that is crucial for spermatogenesis and Leydig cell differentiation, but little is known regarding its influence on gonadal differentiation and development in fish. To understand its function, we cloned and characterized the dhh gene from Cynoglossus semilaevis (csdhh). The full length csdhh cDNA was 2473 bp, including a 1386 bp open reading frame (ORF), a 475 bp 5'-UTR, and a 612 bp 3'-UTR, encoding a predicted protein of 461 amino acid residues. Phylogenetic analysis showed that the putative protein belongs to the hedgehog (HH) family, and contains typical HH-N and HH-C domains. Amino acid sequence analysis revealed that CsDhh shares many features with Dhh analogues in other teleost species. Real-time quantitative PCR showed that csdhh was detected in eight different tissues in male and female tongue sole. During early embryonic development, the relative expression of the csdhh was significantly higher in the neural stage than in other embryonic developmental stages (P < 0.05). csdhh was detected at 20 days after hatching (dah) and at the critical period of male gonadal differentiation (80-95 dah), the relative expression of the csdhh was significantly higher in the male gonads than the female gonads. In 5, 8, and 12 month old gonads, the relative expression of the csdhh was significantly higher in male and pseudo-male than in female fish. The in situ hybridization (ISH) results showed that the hybridization signal was strongly expressed in primary and secondary spermatocytes, spermatids, and sertoli cells of the 1-year-old fish testis, with only weak signal expression in the corresponding ovarian tissue. These results suggest that csdhh is highly conserved in evolution and plays an important role in spermatogenesis in males and pseudo-males.
沙漠刺猬基因（dhh）对于精子发生和莱迪希细胞分化至关重要，但对于其在鱼类性腺分化和发育中的影响知之甚少。为了了解其功能，我们从半滑舌鳎（ Cynoglossus semilaevis ）克隆并表征了 dhh 基因（csdhh）。全长 csdhh cDNA 为 2473bp，包含 1386bp 的开放阅读框（ORF）、475bp 的 5'-UTR 和 612bp 的 3'-UTR，编码一个预测的 461 个氨基酸残基的蛋白质。系统进化分析表明，该假定蛋白属于刺猬（HH）家族，含有典型的 HH-N 和 HH-C 结构域。氨基酸序列分析表明，CsDhh 与其他硬骨鱼类的 Dhh 类似物有许多共同特征。实时定量 PCR 显示，csdhh 在雌雄舌鳎的 8 种不同组织中均有表达。在早期胚胎发育过程中，csdhh 在神经期的相对表达量显著高于其他胚胎发育阶段（P<0.05）。孵化后 20 天（dah）和雄性性腺分化的关键时期（80-95dah）检测到 csdhh，在雄性性腺中的相对表达量显著高于雌性性腺。在 5、8 和 12 月龄的性腺中，csdhh 在雄性和假雄鱼中的相对表达量显著高于雌性鱼。原位杂交（ISH）结果表明，杂交信号在 1 龄鱼睾丸的初级和次级精母细胞、精子细胞和支持细胞中强烈表达，而在相应的卵巢组织中仅表达较弱的信号。这些结果表明，csdhh 在进化上高度保守，在雄性和假雄鱼的精子发生中发挥重要作用。
