Abstract
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.
Highlights
Animal growth and development are regulated by a wide variety of hormones, among which insulin family peptides are important
The present analysis of clustered regularly interspaced palindromic repeats (CRISPR)-Cas9-based gene KO B. mori clearly showed that the BIGFLP gene product regulates the growth and development of the adult organs
The size of appendages such as the antennae and legs was significantly lower in BIGFLP-deficient mutants, suggesting that the BIGFLP gene product plays a role in promoting the growth of these organs
Summary
Animal growth and development are regulated by a wide variety of hormones, among which insulin family peptides are important. An increase in blood glucose levels stimulates the secretion of insulin, which regulates sugar and lipid metabolism in the peripheral tissues[1,2], while another class of insulin family peptides, insulin-like growth factors (IGFs), are known to promote whole-body growth and organ development[3]. ILPs are mainly expressed in the insulin-producing cells (IPCs) in the brain and are released into the haemolymph in response to feeding signals[9,10], whereby they activate insulin/IGF signalling (IIS) in the peripheral tissues to promote cellular growth and metabolism[11]. The inhibition of Dilp[6] expression after pupation by RNA interference reduces the consumption of triacylglycerol and glycogen[15] It appears that Dilp[6] regulates the metabolism and growth of adult organs during development. This study revealed that genetic silencing of the IPCs resulted in a decrease in the growth rate of B. mori larvae, confirming the involvement of ILPs in the regulation of larval growth
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