Molecular and functional characterization of the vitellogenin receptor in oriental river prawn, Macrobrachium nipponense

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• 作者：Hongkun Bai^a ; Hui Qiao^b ; Fajun Li^a ; ^d ; Hongtuo Fu^a ; ^b ; ^{fuht@ffrc.cn" class="auth_mail" title="E-mail the corresponding author} ; Sufei Jiang^b ; Wenyi Zhang^b ; Yuedi Yan^c ; Yiwei Xiong^b ; Shengming Sun^b ; Shubo Jin^a ; ^b ; Yongsheng Gong^b ; Yan Wu^b
• 关键词：Macrobrachium nipponense ; Vitellogenin receptor ; Eyestalk ablation ; RNA interference
• 刊名：Comparative Biochemistry and Physiology - Part A: Molecular & Integrative Physiology
• 出版年：2016
• 出版时间：April 2016
• 年：2016
• 卷：194
• 期：Complete
• 页码：45-55
• 全文大小：3362 K

文摘

A complementary DNA (cDNA) that encodes the vitellogenin receptor (VgR) in the oriental river prawn, Macrobrachium nipponense, was cloned using expressed sequence tag analysis and a rapid amplification of cDNA ends approach. The coding region consists of 5920 base pairs (bp) that encode a 1902 amino acid protein, with a predicted molecular mass of 209 kDa. The coding region is flanked by a 45 bp 5ʹ-untranslated region (UTR) and a 166 bp 3ʹ-UTR. The deduced amino acid sequence of the M. nipponense VgR cDNA had typically conserved domains, such as an extracellular, lipoprotein-binding domain, epidermal growth factor-like and O-glycosylation domains, a transmembrane domain and a short C-terminal, cytosolic tail. Quantitative real-time PCR (qPCR) indicated that Mn-VgR is highly expressed in the female ovary. Expression analysis by qPCR demonstrated the larval and ovarian developmental stage-specific expression pattern. As the ovaries developed, the expression level of Mn-VgR gradually increased during the reproductive cycle (stage I), to reach a peak in stage III. Levels then dropped as a new development cycle was entered after reproduction molting. Eyestalk ablation led to a significant increase in the expression of Mn-VgR during the ovarian development stages (P < 0.05), when compared with the eyestalk-intact group. The investigation revealed that eyestalk ablation initially affected Mn-VgR expression and then influenced vitellogenesis. In adult females, VgR RNA interference (RNAi) dramatically delayed the maturation of the ovary, in accordance with the gonad somatic index. In addition, Mn-VgR RNAi led to vitellin depletion in the oocytes and the accumulation of vitellin in the hepatopancreas.