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Molecular characterization and function analysis of the vitellogenin receptor from the cotton bollworm, Helicoverpa armigera (Hubner) (Lepidoptera, Noctuidae)

机译:棉铃虫Helicoverpa armigera(Hubner)(鳞翅目,夜蛾科)卵黄蛋白原受体的分子特征和功能分析

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摘要

Developing oocytes accumulate plentiful yolk protein during oogenesis through receptor-mediated endocytosis. The vitellogenin receptor (VgR), belonging to the low-density lipoprotein receptor (LDLR) family, regulates the absorption of yolk protein. In this work, the full-length vitellogenin receptor (HaVgR) in the cotton bollworm Helicoverpa armigera was identified, encoding a 1817 residue protein. Sequence alignment revealed that the sequence of HaVgR contained all of the conservative structural motifs of LDLR family members, and phylogenetic analysis indicated that HaVgR had a high identity among Lepidoptera and was distinct from that of other insects. Consistent with other insects, HaVgR was specifically expressed in ovarian tissue. The developmental expression pattern showed that HaVgR was first transcribed in the newly metamorphosed female adults, reached a peak in 2-day-old adults and then declined. Western blot analysis also revealed an ovarian-specific and developing expression pattern, which was consistent with the HaVgR mRNA transcription. Moreover, RNAi-mediated HaVgR knockdown strongly reduced the VgR expression in both the mRNA and protein levels, which inhibited the yolk protein deposition in the ovaries, led to the dramatic accumulation of vitellogenin and the up-regulation of HaVg expression in hemolymph, and eventually resulted in a declined fecundity. Together, all of these findings demonstrate that HaVgR is a specific receptor in uptake and transportation of yolk protein for the maturation of oocytes and that it plays a critical role in female reproduction.
机译:发育中的卵母细胞通过受体介导的内吞作用在卵子形成过程中积累了大量卵黄蛋白。卵黄蛋白原受体(VgR)属于低密度脂蛋白受体(LDLR)家族,可调节卵黄蛋白的吸收。在这项工作中,棉铃虫棉铃虫棉铃虫中的全长卵黄蛋白原受体(HaVgR)被鉴定出来,编码1817个残留蛋白。序列比对表明,HaVgR序列包含LDLR家族成员的所有保守结构基序,系统发育分析表明HaVgR在鳞翅目中具有高度同一性,与其他昆虫截然不同。与其他昆虫一样,HaVgR在卵巢组织中特异性表达。发育表达模式表明,HaVgR首先在新变态的雌性成年人中转录,在2天龄的成年人中达到峰值,然后下降。蛋白质印迹分析还揭示了卵巢特异性和发育中的表达模式,这与HaVgR mRNA转录相符。此外,RNAi介导的HaVgR敲低极大地降低了mRNA和蛋白质水平中的VgR表达,这抑制了卵黄蛋白在卵巢中的沉积,导致卵黄蛋白原的大量积聚和HaVg在血淋巴中的表达上调,最终导致生育力下降。总之,所有这些发现表明,HaVgR是卵黄蛋白摄取和运输中卵母细胞成熟的特定受体,并且在女性生殖中起关键作用。

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