摘要
dazl是重要的母源生殖基因,在鱼类生殖细胞发育和配子产生中具有重要意义。以大黄鱼(Larimichthys crocea)为研究对象,通过同源比对,从大黄鱼基因组中获得dazl基因序列,进行多重比较和进化分析,并借助分子手段半定量RT-PCR检测其在各组织的表达情况,进一步通过原位杂交分析其在性腺生殖细胞的分布。多重比较表明其含有保守的DAZ及DAZL结构域,且进化树显示其与鲈形目鱼类形成一个大支,证实其为大黄鱼dazl基因。半定量RT-PCR进一步分析显示dazl特异性表达于性腺,且精巢表达量高于卵巢。随后,利用原位杂交技术对dazl的时空表达进行定位,发现其在早期生殖细胞表达量较高,随着细胞的发育,表达逐渐减弱,甚至检测不到。dazl在卵巢中主要在卵原和早期卵母细胞中表达,而在精巢中则主要在精原和初级精母细胞中表达,暗示着dazl在雌雄大黄鱼性腺发育过程中发挥重要的作用。研究结果可为探究dazl基因对大黄鱼生殖发育调控机制及两性配子发生奠定基础。
The dazl is an important maternal germ gene,which plays an important role in the development of germ cells and the production of gametes in fish.Larimichthys crocea was selected and the dazl gene sequence was obtained from genome by homologous alignment.Multiple comparisons and evolutionary analysis were conducted with other fish.The dazl gene expression and distribution in various tissues was detected using semi-quantitative RT-PCR and in situ hybridization,respectively.Multiple comparisons indicated that the dazl sequence contained conserved DAZ and DAZL domains and formed a large branch with Perciformes fish as shown in the phylogenetic tree,confirming that it was the dazl gene of L.crocea.Semi-quantitative RT-PCR further analysis showed that dazl was specifically expressed in gonads,and the expression level in testis was higher than that in ovary.In addition,in situ hybridization was used to locate the spatiotemporal expression of dazl.We found that the expression level of dazl was relatively high in early germ cells,and as the cells developed,the expression gradually weakened and could even not be detected;and dazl was mainly expressed in oogonia and early oocytes in the ovary,while in the testis,it was mainly expressed in spermatogonia and primary spermatocytes,suggesting that dazl played an important role in the development of gonads in both male and female individuals.This study laid the foundation for exploring the reproductive development regulation mechanism of the dazl gene and its regulatory role in the formation of gametes in both genders of L.crocea.
作者
李菲艳
曾思源
韩坤煌
孙赵韩
周莉
LI Fei-Yan;ZENG Si-Yuan;HAN Kun-Huang;SUN Zhao-Han;ZHOU Li(College of Ocean Studies,Ningde Normal University,Ningde 352100,China;College of Biological Science and Engineering,Fuzhou University,Fuzhou 350108,China;Ningde Yiye Marine Industry Development Co.,Ltd.,Ningde 352103,China)
出处
《海洋与湖沼》
北大核心
2025年第4期968-975,共8页
Oceanologia Et Limnologia Sinica
基金
宁德师范学院重大科研项目培育计划,2022ZDK02号
宁德师范学院引进人才科研启动项目基金,2022Y06号,2020T03号
福建省自然科学基金项目,2022J05270号,2020J02011号。
关键词
大黄鱼
dazl基因
半定量RT-PCR
原位杂交
发育
large yellow croaker(Larimichthys crocea)
dazl
semi-quantitative RT-PCR
in situ hybridization
development
