| 脊尾白虾VgR基因克隆及其在卵巢发育过程中的表达分析 |
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| 引用本文： | 梁俊平,李健,李吉涛,刘萍,刘德月.脊尾白虾VgR基因克隆及其在卵巢发育过程中的表达分析[J].中国水产科学,2016,23(4):800-812. |
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| 作者姓名： | 梁俊平 李健 李吉涛 刘萍 刘德月 |
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| 作者单位： | 1. 河南师范大学水产学院，河南省水产动物养殖工程技术研究中心，河南新乡 453007; 中国水产科学研究院黄海水产研究所，农业部海洋渔业可持续发展重点实验室，山东青岛 266071;2. 中国水产科学研究院黄海水产研究所，农业部海洋渔业可持续发展重点实验室，山东青岛 266071; 青岛海洋科学与技术国家实验室，海洋渔业科学与食物产出过程功能实验室，山东青岛 266071;3. 中国水产科学研究院黄海水产研究所，农业部海洋渔业可持续发展重点实验室，山东青岛 266071 |
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| 基金项目： | 国家虾产业技术体系(CARS-47);公益性行业(农业)科研专项经费(201103034);青岛海洋科学与技术国家实验室螯山科技创新计划项目(2015ASKJ02) |
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| 摘 要： | 为了解卵黄蛋白原受体(vitellogenin receptor,Vg R)在脊尾白虾卵巢发育中的作用,采用同源克隆和RACE技术,克隆了脊尾白虾Vg R基因全长c DNA序列,用实时荧光定量PCR方法分析了Vg R基因在雌虾不同组织、卵巢发育不同时期的表达特征。结果显示,脊尾白虾Vg R基因全长5892 bp,开放阅读框5661 bp,编码1886个氨基酸。脊尾白虾Vg R具有低密度脂蛋白受体(LDLR)家族典型结构特征,属于LDLR家族,进化上与日本沼虾等甲壳动物Vg R亲缘关系最近,然后与昆虫Vg R分支聚为一支,而与LDLR家族中其他成员亲缘关系较远。脊尾白虾Vg R在各组织中均有表达,但主要在卵巢内表达。随着卵巢的发育,卵巢Vg R表达量逐渐升高,在III期达到最大,与肝胰腺Vg表达量、血液Vg浓度变化趋势相一致;而在卵巢成熟时,卵巢Vg R表达量降到了最低,与肝胰腺Vg表达情况截然相反;排卵后的恢复期,血液中Vg浓度仍维持在较高水平,卵巢Vg R表达量又升至III期水平,同时卵巢Vg表达量也升至最高。由此可见,甲壳动物与昆虫Vg R起源于同一祖先,但在进化上已形成独立一支;脊尾白虾卵巢成熟前,卵巢主要通过Vg R介导作用摄取血液中Vg,以供卵巢快速成熟;卵巢成熟期,肝胰腺呈现补偿性合成Vg,以尽快恢复其营养储备功能;卵巢恢复期,卵巢通过Vg R介导摄入外源Vg和内源合成Vg两种途径,为卵巢二次发育提供营养物质。
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| 关 键 词： | 脊尾白虾 卵黄蛋白原受体 卵黄蛋白原 卵巢发育 组织表达 |
| 修稿时间： | 2016/7/21 0:00:00 |
Molecular cloning of the vitellogenin receptor and its expression during ovarian development of Exopalaemon carinicauda |
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| LIANG Junping,LI Jian,LI Jitao,LIU Ping,LIU Deyue.Molecular cloning of the vitellogenin receptor and its expression during ovarian development of Exopalaemon carinicauda[J].Journal of Fishery Sciences of China,2016,23(4):800-812. |
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| Authors: | LIANG Junping LI Jian LI Jitao LIU Ping LIU Deyue |
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| Institution: | LIANG Junping;LI Jian;LI Jitao;LIU Ping;LIU Deyue;College of Fisheries,Henan Normal University;Engineering Technology Research Center of Henan Province for Aquatic Animal Cultivation;Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences; Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture;Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences;Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture;Functional Laboratory of Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology; |
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| Abstract: | The vitellogenin receptor (VgR) plays an important role in oocyte maturation. VgRs have been studied extensively in various animals from vertebrates to invertebrates, including the chicken,Gallus gallus, fish,On-corhynchus mykiss andOreochromis aureus, insects,Bombyx mori andBlattella germanica, and crustaceans, Macrobrachium rosenbergii andPandalopsis japonica. The aims of this study were to identify and characterize the VgR fromExopalaemon carinicauda, to investigate its expression level during ovarian development, and to de-scribe its interactions with Vg. The full-lengthE. carinicauda VgR cDNA was cloned using degenerated oligonu-cleotide primers and rapid amplification of cDNA ends technology. Quantitative real-time polymerase chain reac-tion analysis was used to quantify relativeVgRexpression levels in different tissues duringE. carinicaudaovarian development. The results showed that the full-lengthE. carinicauda VgR was 5892 bp, containing an open reading frame of 5661 bp, encoding a 1886 amino acid polypeptide. The shrimp VgR contained several conserved domains, such as a ligand-binding domain, an epidermal growth factor-precursor domain, a transmembrane domain, and a cytoplasmic domain; thus, it belongs to the low-density lipoprotein receptor (LDLR) gene family. A phylogenetic analysis revealed that the shrimp VgR was more closely related to VgRs from other crustaceans and insects, rather than to vertebrate VgRs, and was distant from other LDLR members. TheE. carinicauda VgR was expressed in various tissues, and the highest expression was detected in the ovary. OvarianVgR mRNA expression level reached the maximum at stage III of ovarian development and was significantly positively correlated with Vg content in hemolymph andVg expression level in the hepatopancreas before ovarian maturation was completed. The relativeVgR expression level in the ovary reached the minimum at maturation whenVg expression level in the hepatopancreas reached the maximum. Post-spawning Vg concentration remained high when ovarianVg expres-sion level reached the maximum. These results indicate that all VgRs from crustaceans were orthologs of insect VgRs. Vg is synthesized rapidly in the hepatopancreas during maturation to supply a nutrient source for the de-veloping ovary. TheE. carinicauda VgR sequesters Vg in the hemolymph into developing oocytes during ovarian development. Vg was synthesized within and outside the ovary and was transported into the ovary by VgR during the ovarian recovery stage for subsequent ovarian development. |
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| Keywords: | Exopalaemon carinicauda vitellogenin receptor vitellogenin ovarian development tissue expression |
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