The adipokinetic hormone (AKH) of one of the most basal orders of Pterygota: Structure and function of Ephemeroptera AKH

This is the first reported primary sequence of a bioactive peptide isolated from three Ephemeroptera families. Peptides of the adipokinetic hormone (AKH) family from the corpora cardiaca of nymphs of Afronurus spp. (Family: Heptageniidae), Siphlonurus lacustris (Family: Siphlonuridae) and Ephemera danica (Family: Ephemeridae) were investigated functionally in homologous (hypertrehalosaemic activity demonstrated in E. danica nymphs) and heterologous (active in cockroach and locust) bioassays, and structurally by liquid-chromatography coupled with ion trap electrospray ionisation mass spectrometry. All species investigated synthesise the octapeptide code-named Anaim-AKH (pGlu-Val-Asn-Phe-Ser-Pro-Ser-Trp amide). Confirmation of this peptide being present in corpora cardiaca of E. danica nymphs was obtained via reverse phase-high pressure liquid chromatography. Phylogenetically, the presence of only one AKH peptide may constitute a basal condition; all other lower insect orders, e.g. Odonata, Blattodea, Orthoptera, amongst others, have more than one AKH analogue. We propose that Anaim-AKH is the ancestral peptide which may support the Palaeoptera hypothesis that mayflies (Ephemeroptera) and dragonflies (Odonata) form the Palaeoptera clade, the sister group of Neoptera. The structural data cannot, however, shed any light on the phylogenetic scenarios within Ephemeroptera itself. Finally, this study demonstrates the successful use of larvae as an alternative biological source to study neuropeptides in ephemeral, elusive or difficult to obtain adult insects.