Epoxidation of juvenile hormone was a key innovation improving insect reproductive fitness

Methyl farnesoate (MF) plays hormonal regulatory roles in crustaceans. An epoxidated form of MF, known as juvenile hormone (JH), controls metamorphosis and stimulates reproduction in insects. To address the evolutionary significance of MF epoxidation, we generated mosquitoes completely lacking either of the two enzymes that catalyze the last steps of MF/JH biosynthesis and epoxidation, respectively: the JH acid methyltransferase (JHAMT) and the P450 epoxidase CYP15 (EPOX). jhamt−/− larvae lacking both MF and JH died at the onset of metamorphosis. Strikingly, epox−/− mutants, which synthesized MF but no JH, completed the entire life cycle. While epox−/− adults were fertile, the reproductive performance of both sexes was dramatically reduced. Our results suggest that although MF can substitute for the absence of JH in mosquitoes, it is with a significant fitness cost. We propose that MF can fulfill most roles of JH, but its epoxidation to JH was a key innovation providing insects with a reproductive advantage.

Molecular cloning, expression profiling of a carboxylesterase gene and its potential role in methyl farnesoate degradation in Eriocheir sinensis (Brachyura, Varunidae)

In crustaceans, methyl farnesoate (MF) is an important sesquiterpenoid to regulate many physiological processes, especially reproduction and ovarian maturation. In this study, a 1919 bp cDNA of carboxylesterases (Es-CXE6) with some conserved motifs of the CXE multifunctional enzyme family was cloned from Eriocheir sinensis. Tissue and stage-specific expression results suggested that Es-CXE6 expression in hepatopancreas was highest and associated with the haemolymph MF titer. In vitro and in vivo experiments showed that Es-CXE6 expression was significantly upregulated by MF treatment in the hepatopancreas but not in the ovary. Furthermore, an eyestalk ablation experiment showed that Es-CXE6 expression was significantly upregulated on days 1 and 3 post eyestalk ablation in the hepatopancreas. Together, these results indicate that Es-CXE6 may degrade MF in the hepatopancreas in E. sinensis. Our results offer a potential approach to maintain the MF titer at appropriate levels, which has potential applications in crab aquaculture.

A Possible Role of Allatostatin C in Inhibiting Ecdysone Biosynthesis Revealed in the Mud Crab Scylla paramamosain

C-type allatostatins (C-type ASTs) are a family of structurally related neuropeptides found in a wide range of insects and crustaceans. To date, the C-type allatostatin receptor in crustaceans has not been deorphaned, and little is known about its physiological functions. In this study, we aimed to functionally define a C-type ASTs receptor in the mud crab, Scylla paramamosian. We showed that C-type ASTs receptor can be activated by ScypaAST-C peptide in a dose-independent manner and by ScypaAST-CCC peptide in a dose-dependent manner with an IC50 value of 6.683 nM. Subsequently, in vivo and in vitro experiments were performed to investigate the potential roles of ScypaAST-C and ScypaAST-CCC peptides in the regulation of ecdysone (20E) and methyl farnesoate (MF) biosynthesis. The results indicated that ScypaAST-C inhibited biosynthesis of 20E in the Y-organ, whereas ScypaAST-CCC had no effect on the production of 20E. In addition, qRT-PCR showed that both ScypaAST-C and ScypaAST-CCC significantly decreased the level of expression of the MF biosynthetic enzyme gene in the mandibular organ, suggesting that the two neuropeptides have a negative effect on the MF biosynthesis in mandibular organs. In conclusion, this study provided new insight into the physiological roles of AST-C in inhibiting ecdysone biosynthesis. Furthermore, it was revealed that AST-C family peptides might inhibit MF biosynthesis in crustaceans.

Effect of methyl farnesoate and farnesoic acid during 5th zoea and megalopa metamorphosis in the mud crab Scylla paramamosain Estampador, 1950 (Decapoda, Brachyura, Portunidae)

The mud crab Scylla paramamosain is one of the economically important aquaculture species in China. The larval development of the mud crab is characterized by two significant morphological changes, from the 5th zoea (Z5) to the megalopa (M) stage and from the M to the first juvenile crab (C1) stage. In this study, we found that methyl farnesoate (MF) could prohibit the Z5 to M metamorphosis in a concentration-dependent manner, and that a concentration of 10 μM MF could completely prohibit the Z5 metamorphosis. Farnesoic acid (FA) could also prohibit the Z5 metamorphosis, but its effects seemed to be concentration-independent. In addition, MF could delay rather than prohibit the M to C1 metamorphosis, while FA had no effect on the M to C1 metamorphosis at all. To summarize, it is hypothesized that either absence of MF and FA, or at least very low levels of these substances, might be necessary for a successful Z5 to M metamorphosis.

Juvenile hormone III skipped bisepoxide is widespread in true bugs (Hemiptera: Heteroptera)

Juvenile hormone (JH) plays important roles in almost every aspect of insect development and reproduction. JHs are a group of acyclic sesquiterpenoids, and their farnesol backbone has been chemically modified to generate a homologous series of hormones in some insect lineages. JH III (methyl farnesoate, 10,11-epoxide) is the most common JH in insects, but Lepidoptera (butterflies and moths) and ‘higher’ Diptera (suborder: Brachycera; flies) have developed their own unique JHs. Although JH was first proposed in the hemipteran suborder Heteroptera (true bugs), the chemical identity of the heteropteran JH was only recently determined. Furthermore, recent studies revealed the presence of a novel JH, JH III skipped bisepoxide (JHSB 3 ), in some heteropterans, but its taxonomic distribution remains largely unknown. In the present study, we investigated JHSB 3 production in 31 heteropteran species, covering almost all heteropteran lineages, through ultra-performance liquid chromatography coupled with tandem mass spectrometry. We found that all of the focal species produced JHSB 3 , indicating that JHSB 3 is widespread in heteropteran bugs and the evolutionary occurrence of JHSB 3 ascends to the common ancestor of Heteroptera.