青鳉精巢gdnfa与gdnfb的细胞表达模式及视黄酸和11-酮基睾酮对其差异调控作用  

Differential expression patterns of gdnfa and gdnfb in the testis of medaka(Oryzias latipes)and their regulation by retinoic acid and 11-ketotestosterone

作　　者：屈锡梅 王园 赵长乐 刘磊 陶文静 王德寿 魏静 QU Ximei;WANG Yuan;ZHAO Changle;LIU Lei;TAO Wenjing;WANG Deshou;WEI Jing(Integrative Science Center of Germplasm Creation in Western China(Chongqing)Science City,Key Laboratory of Freshwater Fish Reproduction and Development Ministry of Education,School of Life Sciences,Southwest University,Chongqing 400715,China;Chengdu Kangnuoxing Biopharmaceutical Technology Co.,Ltd.,Chengdu 610219,China;Guizhou Hongcai Junong Investment Co.,Ltd.,Liupanshui 553500,China)

机构地区：[1]西部(重庆)科学城种质创制大科学中心,西南大学生命科学学院,淡水鱼类资源与生殖发育教育部重点实验室,重庆400715 [2]成都康诺行生物医药科技有限公司,四川成都610219 [3]贵州宏财聚农投资有限责任公司,贵州六盘水553500

出　　处：《水产学报》2025年第1期64-74,共11页Journal of Fisheries of China

基　　金：国家自然科学基金(31972776,32172969,31972778,32172953);重庆市自然科学基金(cstc2020jcyjmsxmX1045)。

摘　　要：【目的】探究青鳉胶质细胞源神经营养因子(GDNF)在精巢中的细胞表达模式,及视黄酸(RA)和11-酮基睾酮(11-KT)对其表达调控作用。【方法】通过荧光原位杂交(FISH)检测了青鳉gdnf的2个复制基因(即gdnfa与gdnfb)在精巢中的细胞表达模式,并通过实时定量聚合酶链式反应(qRT-PCR)、5′端上游序列转录活性分析等从组织、细胞及分子水平探究了精巢分化发育重要调控因子RA和11-KT对二者的表达调控作用。【结果】在精巢中,gdnfa主要表达于体细胞,而gdnfb除表达于体细胞外,在生殖细胞中也有明显表达。不同浓度RA和11-KT处理体外培养的青鳉精巢组织及其传代培养体细胞MTS1,结果表明,二者均可显著下调gdnfa的表达,上调gdnfb的表达。分别用gdnfa与gdnfb不同截短5′端上游序列的荧光素酶报告载体转染MTS1,结果显示,RA处理可降低gdnfa的多个截短5′端上游序列荧光素酶活性,增强gdnfb的多个截短5′端上游序列荧光素酶活性,11-KT处理的结果与此基本相似。【结论】青鳉gdnfa与gdnfb在精巢中具有差异的细胞表达模式,同时二者在组织、细胞及分子水平均受到了RA和11-KT的差异化调控。本研究深化了对鱼类gdnf的2个复制基因的细胞表达模式及其调控的深入认识,为其进一步生物学功能研究奠定了重要基础。Glial cell line-derived neurotrophic factor(GDNF)is a key factor mediating the self-renewal and maintenance of spermatogonial stem cells(SSCs).Our previous research indicated that there were two gdnf genes in medaka(Oryzias latipes),namely gdnfa and gdnfb,both of which were expressed in the testis and play important roles in the spermatogonial stem cell line SG3.However,the cellular expression patterns of gdnfa and gdnfb in the testis,as well as regulation of their expressions by retinoic acid(RA)and androgen,remain unclear.In this study,the cellular expression patterns of medaka gdnfa and gdnfb in the testis were detected using fluorescence in situ hybridization(FISH).Furthermore,the regulation of RA and 11-ketotestosterone(11-KT)on the expression of gdnfa and gdnfb was investigated at the tissue,cell,and molecular levels through real-time quantitative polymerase chain reaction(qRT-PCR)and transcriptional activity analyses of their 5′upstream sequences.These results indicated that in medaka testicular sections,gdnfa was primarily expressed in somatic cells,while gdnfb was expressed in both germ cells and somatic cells.In testicular organ culture and somatic cells MTS1 derived from adult medaka testis,RA and 11-KT at different concentrations significantly down-regulated the expression of gdnfa,whereas up-regulated the expression of gdnfb.moreover,RA and 11-KT treatment reduced the luciferase activity of different truncated 5′upstream sequences of gdnfa,but enhanced the luciferase activity of different truncated 5′upstream sequences of gdnfb.Collectively,medaka gdnfa and gdnfb exhibit differential cellular expression patterns in the testis and are differentially regulated by RA and 11-KT at the tissue,cell,and molecular levels.This study deepens our understanding of the cellular expression patterns and regulation of medaka gdnfa and gdnfb,and lays an important foundation for further research on their biological functions.

关 键 词：青鳉 胶质细胞源神经营养因子 视黄酸 11-酮基睾酮 精巢 体细胞 

分 类 号：S917.4[农业科学—水产科学]