Characterization and expression analysis of adipokinetic hormone and its receptor in eusocial aphid Pseudoregma bambucicola.

Aphids display an extraordinary phenotypic plasticity ranging from widespread reproductive and wing polyphenisms to the occurrence of sterile or subfertile soldier morphs restricted to eusocial species of the subfamilies Eriosomatinae and Hormaphidinae. Individual morphs are specialized by their behavior, anatomy, and physiology to perform different roles in aphid societies at different stages of the life cycle. The capacity of the insects to cope with environmental stressors is under the control of a group of neuropeptides of the adipokinetic hormone/red pigment-concentrating hormone family (AKH/RPCH) that bind to a specific receptor (AKHR). Here, we describe the molecular characteristics of AKH and AKHR in the eusocial aphid Pseudoregma bambucicola. The sequence of the bioactive AKH decapeptide and the intron position in P. bambucicola AKH preprohormone were found to be identical to those in a phylogenetically distant aphid Dreyfusia spp. (Adelgidae). We detected four transcript variants of AKHR that are translated into three protein isoforms. Further, we analyzed AKH/AKHR expression in different tissues and insects of different castes. In wingless females, a remarkable amount of AKH mRNA was only expressed in the heads. In contrast, AKHR transcript levels increased in the order gut<ovary<fat body<head. In aphids from both the primary and secondary hosts (Styrax suberifolia and Bambusa spp., respectively), the highest AKH expression levels were recorded in winged, migratory females and soldiers, whereas reduced levels were found in wingless, sedentary females that are functionally oriented to reproduction. The highest AKHR expression was found in soldiers in gall-dwelling populations, whereas in bamboo colonies the highest transcript level was detected in winged females. We propose a possible explanation for the correlation between AKH and AKHR transcript levels and task partitioning among individual forms in P. bambucicola colonies.

Jedličková V ，Jedlička P ，Lee HJ 《-》

The adipokinetic hormone system in Culicinae (Diptera: Culicidae): molecular identification and characterization of two adipokinetic hormone (AKH) precursors from Aedes aegypti and Culex pipiens and two putative AKH receptor variants from A. aegypti.

Kaufmann C ，Merzendorfer H ，Gäde G 《-》

Functional characterization of the adipokinetic hormone in the pea aphid, Acyrthosiphon pisum.

Aphids are important plant phloem-sucking pests and detailed knowledge about the hormonal control of their metabolism can potentially contribute to the development of methods for their management. The insect metabolism is predominantly controlled by neuropeptides belonging to the adipokinetic hormone/red pigment-concentrating hormone family (AKH/RPCH). The main goal of this study was to obtain the sequence of AKH transcripts and analyze its expression in all polyphenic female forms of the pea aphid, Acyrthosiphon pisum. The neuropeptide is expressed in the brain of all female forms and in the ovaries of the both (wingless and winged) parthenogenetic forms. The form of active Acypi-AKH decapeptide was confirmed by the LC/MS and +ESI tandem mass spectrometry. The highest relative amount of Acypi-AKH was recorded in winged virginoparae. Furthermore, a potential role of this hormone when directly applied to the aphid was studied as well. Interestingly, no significant increase of trehalose in the wingless virginoparae after application of synthetic Acypi-AKH was detected. Yet this treatment did affect the level of protective polyol (mannitol) and furthermore led to increased activity of the detoxification enzyme glutathione S-transferase. The possible physiological function of AKH in A. pisum under the stress conditions is discussed.

Jedlička P ，Steinbauerová V ，Simek P ，Zahradníčková H ... -  《-》

Adipokinetic hormone signalling system in the Chagas disease vector, Rhodnius prolixus.

Neuropeptides and their G protein-coupled receptors are widespread throughout Metazoa and in several cases, clear orthologues can be identified in both protostomes and deuterostomes. One such neuropeptide is the insect adipokinetic hormone (AKH), which is related to the mammalian gonadotropin-releasing hormone. AKH has been studied extensively and is known to mobilize lipid, carbohydrates and proline for energy-consuming activities such as flight. In order to determine the possible roles for this signalling system in Rhodnius prolixus, we isolated the cDNA sequences encoding R. prolixus AKH (Rhopr-AKH) and its receptor (Rhopr-AKHR). We also examined their spatial expression pattern using quantitative PCR. Our expression analysis indicates that Rhopr-AKH is only expressed in the corpus cardiacum of fifth-instars and adults. Rhopr-AKHR, by contrast, is expressed in several peripheral tissues including the fat body. The expression of the receptor in the fat body suggests that AKH is involved in lipid mobilization, which was confirmed by knockdown of Rhopr-AKHR via RNA interference. Adult males that had been injected with double-stranded RNA (dsRNA) for Rhopr-AKHR exhibited increased lipid content in the fat body and decreased lipid levels in the haemolymph. Moreover, injection of Rhopr-AKH in Rhopr-AKHR dsRNA-treated males failed to elevate haemolymph lipid levels, confirming that this is indeed the receptor for Rhopr-AKH.

Zandawala M ，Hamoudi Z ，Lange AB ，Orchard I ... -  《-》

Adipokinetic hormone receptor gene identification and its role in triacylglycerol mobilization and sexual behavior in the oriental fruit fly (Bactrocera dorsalis).

Energy homeostasis requires continuous compensation for fluctuations in energy expenditure and availability of food resources. In insects, energy mobilization is under control of the adipokinetic hormone (AKH) where it is regulating the nutritional status by supporting the mobilization of lipids. In this study, we characterized the gene coding for the AKH receptor (AKHR) and investigated its function in the oriental fruit fly (Bactrocera dorsalis) that is economically one of the most important pest insects of tropical and subtropical fruit. Bacdo-AKHR is a typical G protein-coupled receptor (GPCR) and phylogenetic analysis confirmed that Bacdo-AKHR is closely related to insect AKHRs from other species. When expressed in Chinese hamster ovary (CHO) cells, Bacdo-AKHR exhibited a high sensitivity and selectivity for AKH peptide (EC50 = 19.3 nM). Using qPCR, the developmental stage and tissue-specific expression profiles demonstrated that Bacdo-AKHR was highly expressed in both the larval and adult stages, and also specifically in the fat body and midgut of the adult with no difference in sex. To investigate the role of AKHR in B. dorsalis, RNAi assays were performed with dsRNA against Bacdo-AKHR in adult flies of both sexes and under starvation and feeding condition. As major results, the knockdown of this gene resulted in triacylglycerol (TAG) accumulation. With RNAi-males, we observed a severe decrease in their sexual courtship activity when starved, but there was a partial rescue in copulation when refed. Also in RNAi-males, the tethered-flight duration declined compared with the control group when starved, which is confirming the dependency on energy metabolism. In RNAi-females, the sexual behavior was not affected, but their fecundity was decreased. Our findings indicate an interesting role of AKHR in the sexual behavior of males specifically. The effects are associated with TAG accumulation, and we also reported that the conserved role of AKH-mediated system in B. dorsalis is nutritional state-dependent. Hence, we provided further understanding on the multiple functions of AKH/AKHR in B. dorsalis.

Hou QL ，Chen EH ，Jiang HB ，Wei DD ，Gui SH ，Wang JJ ，Smagghe G ... -  《-》