Metabolomics reveals changes in metabolite profiles due to growth and metamorphosis during the ontogeny of the northern damselfly.

Many insects have complex life cycles where a drastic ontogenetic change happens between the larval stages and the adult stage, i.e. metamorphosis. Damselflies (order Odonata, suborder Zygoptera) are widely distributed and ecologically important semi-aquatic insects with a complex life cycle. Phenotypic changes over damselfly ontogeny have been documented, however, if and how metabolite profiles are also changing is currently unknown. Here we used a metabolomics methodology to gain insights into the metabolic changes during the life cycle of the Northern damselfly (Coenagrion hastulatum). Hatchlings of wild-caught damselflies were reared in the laboratory and metabolomics analyses using liquid chromatography and gas chromatography coupled to mass spectrometry were carried out at three larval stages and on adult damselflies. Additionally, a subset of larvae was exposed to wastewater effluent to assess how metabolite profiles responded to an environmental stressor. A total of 212 compounds belonging to several classes (e.g. amino acids, fatty acids, sugars) were annotated. Across metamorphosis, we found that damselflies shifted from protein catabolism to lipid catabolism. Wastewater effluent exposure resulted in ontogenetic stage-dependent changes of individual metabolites, but not to a marked extent. Overall, our study is one of the first to describe changes of metabolite profiles during ontogeny of an insect, and it provides a first step towards a greater understanding of the physiological changes occurring during general insect-but especially damselfly-ontogeny.