Department of Integrated Biosciences, Graduate School of Frontier Sciences, The University of Tokyo, Kashiwa, Chiba, 277-8562, Japan.
Faculty of Textile Science and Technology, Shinshu University, Ueda, 386-8567, Japan.
Sci Rep. 2019 Dec 5;9(1):18446. doi: 10.1038/s41598-019-54962-w.
Insulin family peptides are known to be key regulators of growth and metabolism in insects and vertebrates. Insects have two types of insulin family peptides: insulin-like peptides and insulin-like growth factor (IGF)-like peptides (IGFLPs). We recently demonstrated that an IGFLP in the silkmoth, Bombyx mori (BIGFLP) promotes the growth of the genital imaginal disc ex vivo. However, the role of BIGFLP in the regulation of insect growth remains unclear because no in vivo study has been performed. Therefore, we analysed the functions of BIGFLP in vivo by constructing BIGFLP knock-out (KO) B. mori using the clustered regularly interspaced palindromic repeats (CRISPR) and CRISPR-associated protein 9 (CRISPR-Cas9) system. The KO moths exhibited decreased body weights and size of the appendages compared wild-type (wt) moths. Interestingly, KO females also had drastically lower ovary weights and number of eggs than wt females. However, mutant ovaries that were transplanted into wt host pupae reached a similar weight to wt ovaries that were transplanted into the wt hosts, suggesting that IGFLP in the haemolymph promotes ovarian development. These findings show that BIGFLP regulates the growth and development of adult organs, particularly the ovaries, in B. mori.
胰岛素家族肽是昆虫和脊椎动物生长和代谢的关键调节剂。昆虫有两种类型的胰岛素家族肽:胰岛素样肽和胰岛素样生长因子(IGF)样肽(IGFLP)。我们最近证明,家蚕(Bombyx mori)中的一种 IGFLP(BIGFLP)促进了生殖器 imaginal 盘的体外生长。然而,由于没有进行体内研究,BIGFLP 在昆虫生长调节中的作用尚不清楚。因此,我们使用簇状规则间隔的短回文重复序列(CRISPR)和 CRISPR 相关蛋白 9(CRISPR-Cas9)系统构建了 BIGFLP 敲除(KO)家蚕,从而在体内分析了 BIGFLP 的功能。与野生型(wt)家蚕相比,KO 家蚕的体重和附肢大小降低。有趣的是,KO 雌蛾的卵巢重量和产卵数也明显低于 wt 雌蛾。然而,移植到 wt 宿主蛹中的突变卵巢的重量与移植到 wt 宿主的 wt 卵巢相似,这表明血液中的 IGFLP 促进了卵巢发育。这些发现表明,BIGFLP 在家蚕中调节成年器官的生长和发育,特别是卵巢。