西北农林科技大学学报（自然科学版）2026，Vol.54Issue(4)：17-25,9.DOI:10.13207/j.jnwafu.2026.04.003
大鳍鳠dnd基因的克隆与表达特征分析
Cloning and expression analysis of the dnd gene in Hemibagrus macropterus
摘要
Abstract
[Objective]This study aimed to investigate the expression patterns of the germ cell-specific marker gene dead end(dnd)during gonadal development in Hemibagrus macropterus.[Method]Tissues in-cluding the heart,liver,spleen,kidney,intestine,brain,gills,and gonads(ovary and testis)were collected from 1-,2-,and 4-year-old male and female H.macropterus.Total RNA was extracted from the ovaries of 4-year-old H.macropterus,which was reverse-transcribed into cDNA,and the full-length cDNA sequence of H.mac-ropterus dnd(Hmdnd)was cloned using RACE technology.Subsequently,sequence analysis was conducted.Using β-actin as the reference gene,real-time quantitative PCR(RT-qPCR)was employed to quantify dnd ex-pression in various tissues of 1-year-old H.macropterus and in gonads at different developmental stages.RNA in situ hybridization(ISH)was applied to localize dnd mRNA within germ cells.[Result]The full-length Hmdnd cDNA sequence was 2 035 bp(GenBank accession:PQ626051),containing an open reading frame(ORF)of 1 131 bp that encodes 376 amino acids.Sequence alignment and phylogenetic analysis revealed the highest protein sequence identity(79.24%)between H.macropterus and Pelteobagrus fulvidraco(yellow cat-fish).RT-qPCR demonstrated that Hmdnd was exclusively expressed in gonads,with significantly higher levels in ovaries than in testes,and no expression in somatic tissues.Hmdnd expression declined markedly during gam-ete maturation.RNA in ISH of gonadal sections revealed that Hmdnd mRNA was predominantly localized in the cytoplasm of germ cells.In ovaries,Hmdnd expression increased sharply as primary oocytes developed from stage Ⅰ to stage Ⅱ,but dropped to stage-Ⅰ-equivalent levels in stage Ⅲ primary oocytes.In testes,the stron-gest Hmdnd signals were detected in spermatogonia and diminished progressively during spermatogenesis,with no expression observed in spermatozoa.[Conclusion]Hmdnd is likely to play a critical role in the early gonadal development of H.macropterus.关键词
大鳍鳠/dnd基因/生殖细胞/性腺发育Key words
Hemibagrus macropterus/dnd gene/germ cell/gonadal development分类
农业科技引用本文复制引用
余乐,王锐,叶欢,岳华梅,谢勇,易沫,陈元坤,李创举..大鳍鳠dnd基因的克隆与表达特征分析[J].西北农林科技大学学报（自然科学版）,2026,54(4):17-25,9.基金项目
重庆市科研机构绩效激励引导专项(CSTB2023JXJL-YFX0084) （CSTB2023JXJL-YFX0084）
重庆市财政项目(C20240103) （C20240103）
国家重点研发计划项目(2022YFD2400101) （2022YFD2400101）
中央级公益性科研院所基本科研业务费专项资金项目(2023TD23) （2023TD23）
