半滑舌鳎(Cynoglossus semilaevis)Patched 1基因的克隆和表达分析

Hedgehog(Hh)信号通路在脊椎动物和非脊椎动物发育过程中细胞信号传导方面发挥重要的作用。Patched 1 (Ptc1)基因是Hh的受体,通过和Hh配体结合调节Hh信号通路。本研究克隆和鉴定了半滑舌鳎的Ptc1基因(csptc1),该基因cDNA全长为5212 nt,编码蛋白包括1543个氨基酸残基,序列比对和进化分析显示该蛋白在进化过程中较为保守。荧光定量显示该基因在脑,肝脏,心脏,鳃,肠,脾脏、肾脏和性腺组织中普遍表达,在精巢中的表达水平显著高于卵巢。精巢中的原位杂交信号主要位于该基因主要在初级精母细胞,次级精母细胞和支持细胞中,而在卵母细胞中的信号较弱。Csptc1基因在卵巢中的甲基化水平高于在雄鱼和伪雄鱼精巢中。这一研究结果可能说明csptc1基因参与了 Desert Hedgehog (DHH)基因维持雄鱼和伪雄鱼的生殖细胞系以及精子发生等生物学过程。

Molecular characterization and expression analysis of Patched 1 gene in the half-smooth tongue sole (Cynoglossus semilaevis)

The signaling molecule hedgehog (Hh) is essential for cellular signaling required for body pattern formation in embryonic and individual development in vertebrate and invertebrates. Patched 1 (Ptc1), a receptor of Hh, mediates the activity of Hh by binging the Hh ligands. In this study, the Ptc1 of half-smooth tongue sole Cynoglossus semilaevis (CsPtc1) was cloned and characterized. The full-length of CsPtc1 cDNA is 5 212 nucleotides (nt), and encodes a protein with 1 543 amino acid residues. CsPtc1 shared many features with Ptc1 of other telesot species. Real-time quantitative PCR revealed CsPtc1 mRNA expressed in brain, liver, heart, gill, intestines, spleen, gonad and kidney, with expression level in testis significantly higher than in ovary. In testis, hybridization signals were mainly detected in primary spermatocytes, secondary spermatocytes, and sertoli cells, while weak signals were found in oocytes. Correspondingly, the degree of methylation is higher in female than in male and pseudo-male. Results indicate that CsPtc1 may be involved in Desert Hedgehog (DHH) maintenance of the male and pseudo-male germ line and spermatogenesis.