Significance: The juvenile hormone (JH) plays a key role in regulating insect reproduction. Previous studies have demonstrated how JH activates vitellogenin production in the fat body and stimulates vitellogenin uptake by maturing oocytes. Our studies show that, in Drosophila , JH signaling induces laminin expression in ovarian muscle cells and collagen IV expression in the adult fat body. Laminin locally and collagen IV remotely contribute to the assembly of ovarian muscle extracellular matrix (ECM). The ECM components are indispensable for ovarian muscle contraction that externally generates a mechanical force to promote ovulation and maintain egg shape. This study reveals an essential role of JH in the regulation of insect reproduction.

In the field of insect endocrinology, juvenile hormone (JH) is one of the most wondrous entomological terms. As a unique sesquiterpenoid hormone produced and released by the endocrine gland, corpus allatum (CA), JH is a critical regulator in multiple developmental and physiological processes, such as metamorphosis, reproduction, and behavior. Benefited from the precise genetic interventions and simplicity, the fruit fly, Drosophila melanogaster, is an indispensable model in JH studies. This review is aimed to present the regulatory factors on JH biosynthesis and an overview of the regulatory roles of JH in Drosophila. The future directions of JH studies are also discussed, and a few hot spots are highlighted.

In insects, 20-hydroxyecdysone (20E) limits the growth period by triggering developmental transitions; 20E also modulates the growth rate by antagonizing insulin/insulin-like growth factor signaling (IIS). Previous work has shown that 20E cross-talks with IIS, but the underlying molecular mechanisms are not fully understood. Here we found that, in both the silkworm Bombyx mori and the fruit fly Drosophila melanogaster, 20E antagonized IIS through the AMP-activated protein kinase (AMPK)-protein phosphatase 2A (PP2A) axis in the fat body and suppressed the growth rate. During Bombyx larval molt or Drosophila pupariation, high levels of 20E activate AMPK, a molecular sensor that maintains energy homeostasis in the insect fat body. In turn, AMPK activates PP2A, which further dephosphorylates insulin receptor and protein kinase B (AKT), thus inhibiting IIS. Activation of the AMPK-PP2A axis and inhibition of IIS in the Drosophila fat body reduced food consumption, resulting in the restriction of growth rate and body weight. Overall, our study revealed an important mechanism by which 20E antagonizes IIS in the insect fat body to restrict the larval growth rate, thereby expanding our understanding of the comprehensive regulatory mechanisms of final body size in animals. Significance Final body size determination in animals remains a long-standing puzzle in the field of developmental biology. In insects, insulin/insulin-like growth factor signaling (IIS) and the steroid hormone 20-hydroxyecdysone (20E) are mainly responsible for the growth rate and growth period, respectively, and consequently the final body size. Here, we report that 20E activates AMPK by up-regulating its gene expression and inducing sugar starvation. In turn, AMPK activates protein phosphatase 2A, which further dephosphorylates and inactivates key components in IIS in the fat body, and eventually suppresses the larval growth rate. These data answer the question of how 20E antagonizes IIS and modulates growth rate.

Juvenile hormone (JH) regulates insect development and reproduction through both intracellular and membrane signaling, and the two pathways might crosstalk with each other. Recent studies have reported that JH membrane signaling induces phosphorylation of the JH intracellular receptor Met, thus enhancing its transcriptional activity. To gain more insights into JH-induced Met phosphorylation, we here performed phosphoproteomics to identify potential phosphorylation sites of Met and its paralog Germ-cell expressed (Gce) in Drosophila Kc cells. In vitro experiments demonstrate that JH-induced phosphorylation sites in the basic helix-loop-helix (bHLH) domain, but not in the Per-Arnt-Sim-B (PAS-B) domain, are required for maximization of Met transcriptional activity. Moreover, phosphoproteomics analysis reveale that JH also induces the phosphorylation of Hsp83, a chaperone protein involved in JH-activated Met nuclear import. The JH-induced Hsp83 phosphorylation at S219 facilitates Hsp83-Met binding, thus promoting Met nuclear import and its transcription. By using proteomics, subcellular distribution, and co-immunoprecipitation approaches, we further characterized 14-3-3 proteins as negative regulators of Met nuclear import through physical interaction with Hsp83. These results show that JH membrane signaling induces phosphorylation of the key components in JH intracellular signaling, such as Met and Hsp83, and consequently facilitating JH intracellular signaling.