Insulin receptor-mediated nutritional signalling regulates juvenile hormone biosynthesis and vitellogenin production in the German cockroach.

Female reproductive processes, which comprise, amongst others, the synthesis of yolk proteins and the endocrine mechanisms which regulate this synthesis, need a considerable amount of energy and resources. The role of communicating that the required nutritional status has been attained is carried out by nutritional signalling pathways and, in particular, by the insulin receptor (InR) pathway. In the present study, using the German cockroach, Blattella germanica, as a model, we analysed the role of InR in different processes, but mainly those related to juvenile hormone (JH) synthesis and vitellogenin production. We first cloned the InR cDNA from B. germanica (BgInR) and then determined that its expression levels were constant in corpora allata and fat body during the first female gonadotrophic cycle. Results showed that the observed increase in BgInR mRNA in fat body from starved compared to fed females was abolished in those females treated with systemic RNAi in vivo against the transcription factor BgFoxO. RNAi-mediated BgInR knockdown during the final two nymphal stages produced significant delays in the moults, together with smaller adult females which could not spread the fore- and hindwings properly. In addition, BgInR knockdown led to a severe inhibition of juvenile hormone synthesis in adult female corpora allata, with a concomitant reduction of mRNA levels corresponding to 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) synthase-1, HMG-CoA synthase-2, HMG-CoA reductase and methyl farnesoate epoxidase. BgInR RNAi treatment also reduced fat body vitellogenin mRNA and oocyte growth. Our results show that BgInR knockdown produces similar phenotypes to those obtained in starved females in terms of corpora allata activity and vitellogenin synthesis, and indicate that the InR pathway mediates the activation of JH biosynthesis and vitellogenin production elicited by nutrition signalling.

FoxO is required for the activation of hypertrehalosemic hormone expression in cockroaches.

BACKGROUND: FoxO proteins are a subgroup of the Forkhead-box family of transcription factors, which function as the main transcriptional effectors of the insulin receptor pathway. This pathway, activated by the binding of insulin or IGFs (or insect insulin-like peptides), promotes the phosphorylation and inactivation of FoxO because of its export from the nucleus to the cytoplasm. The homolog of FoxO in the cockroach Blattella germanica works in a situation of nutrient shortage by inhibiting the endocrine induction of reproduction. METHODS: Using Blattella germanica as a model, we studied the functions of FoxO using RNA interference methodologies. We analyzed the mRNA levels of hypertrehalosemic hormone (HTH) and genes related to lipolysis, glycogenolysis and gluconeogenesis and quantified triacylglycerides, glycogen and trehalose. RESULTS: FoxO knockdown eliminates the starvation-induced expression of HTH in the corpora cardiaca. In addition, FoxO knockdown prevents the activation of the expression of Brummer lipase, glycogen phosphorylase and phosphoenolpyruvate carboxylase in the fat body of starved females. CONCLUSIONS: Starvation-induced activation of FoxO stimulates the transcription of different genes related to catabolic processes, including HTH and genes involved in lipolysis, glycogenolysis and gluconeogenesis. GENERAL SIGNIFICANCE: Our results show conservation in the action of the transcription factor FoxO in the activation of catabolic processes from basal insects to vertebrates. The results also describe a new and essentially different mode of action of transcription factor FoxO, which works through the activation of neuropeptide HTH expression, which will subsequently produce its own catabolic stimulatory function.

FoxO inhibits juvenile hormone biosynthesis and vitellogenin production in the German cockroach.

The transcription factor Forkhead-box O (FoxO) is the main transcriptional effector of the Insulin Receptor/Phosphatidylinositol 3-kinase (InR/PI3K) pathway. In a situation of nutrient restriction, the pathway is inactive and FoxO translocates to the nucleus to exert its transcriptional action. In starved females of the cockroach Blattella germanica, the reproductive processes, and in particular the synthesis of juvenile hormone in the corpora allata and that of vitellogenin in the fat body, are arrested. In the present report we examine the possible role of FoxO in the transduction of the nutritional signals to these reproductive events. We first cloned FoxO cDNA from B. germanica (BgFoxO), and showed that its expression is not nutritionally regulated. BgFoxO knockdown using systemic RNAi in vivo in starved females elicited an increase of juvenile hormone biosynthesis, although without modifying mRNA levels of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) synthase-1, HMG-CoA synthase-2, HMG-CoA reductase or methyl farnesoate epoxidase (CYP15A1) in corpora allata. In addition, BgFoxO RNAi treatment produced a remarkable increase of vitellogenin mRNA levels in fat body and of vitellogenin protein in the haemolymph. Our results indicate that BgFoxO plays an inhibitory role on juvenile hormone biosynthesis and vitellogenin production in a situation of nutrient shortage.