Systemic RNAi of the cockroach vitellogenin receptor results in a phenotype similar to that of the Drosophila yolkless mutant
Systemic RNAi of the cockroach vitellogenin receptor results in a phenotype similar to that of the Drosophila yolkless mutant
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DOI：
10.1111/j.1742-4658.2005.05066.x
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发表时间：
2006-01-01
期刊：
影响因子：
5.4
通讯作者：
Bellés, X
中科院分区：
文献类型：
--
作者：
Ciudad, L;Piulachs, MD;Bellés, X
卵黄发生是卵生生殖过程中调控最严格的过程之一，卵黄原蛋白通过卵黄原蛋白受体（VgR）介导的内吞作用进入卵母细胞。本文报道了德国小蠊VgR基因cDNA的克隆，并首次利用RNA干扰（RNAi）技术对VgR基因进行了功能分析。我们的特点是VgR，VgR的mRNA和蛋白质的表达模式，在成年前和成年阶段的这种蟑螂，以及VgR的免疫定位在卵巢管，属于panoistic型。然后，我们使用RNAi技术特异性地破坏VgR基因功能。敲低VgR表达导致的表型，其特征在于低卵黄含量的卵巢和高卵黄蛋白原浓度的血淋巴。这种表型相当于黑腹果蝇的无卵黄突变体，其yl（VgR）基因被破坏。这些结果还开辟了这样一个前景，即可以通过系统性RNAi在这种基础物种中对发育基因进行功能分析。
During vitellogenesis, one of the most tightly regulated processes in oviparous reproduction, vitellogenins are incorporated into the oocyte through vitellogenin receptor (VgR)-mediated endocytosis. In this paper, we report the cloning of the VgR cDNA from Blattella germanica, as well as the first functional analysis of VgR following an RNA interference (RNAi) approach. We characterized the VgR, VgR mRNA and protein expression patterns in pre-adult and adult stages of this cockroach, as well as VgR immunolocalization in ovarioles, belonging to the panoistic type. We then specifically disrupted VgR gene function using RNAi techniques. Knockdown of VgR expression led to a phenotype characterized by low yolk content in the ovary and high vitellogenin concentration in the haemolymph. This phenotype is equivalent to that of the yolkless mutant of Drosophila melanogaster, which have the yl (VgR) gene disrupted. The results additionally open the perspective that development genes can be functionally analyzed via systemic RNAi in this basal species.