Hyperthermia induced atrial natriuretic peptide expression and deviant heart development in Atlantic salmon Salmo salar embryos

Heart abnormalities are increasingly recognized as a problem in salmon aquaculture. Fish in early life-stages are particularly susceptible to teratogens, including elevated water temperature. Recently, heat-induced mRNA expression of the cardiac hormone atrial natriuretic peptide (ANP), which is known to be involved in modulation of cardiac growth and regulation of cardiac homeostasis, was demonstrated in Atlantic salmon (Salmo salar) embryos by RAP-PCR. The relation between heat sensitive ANP expression and heart abnormalities was explored in two experiments. In an experiment with short-term exposure, salmon eggs were heat shocked at 16 °C at eight different embryonic stages from gastrulation till completion of somitogenesis. The RT-PCR results showed that the ANP mRNA expression was down-regulated at the onset of heart formation at the gastrula stage, while the transcription became heat inducible from the fusioning of the heart tube around the 15th–20th somite stage and onwards. This was confirmed by whole-mount in situ hybridization, which also showed that ANP is exclusively expressed in the heart of Atlantic salmon embryos. In a second long-term experiment, salmon embryos were incubated at either 10 °C (high temperature) or 8 °C (controls) from fertilization till first feeding, and subsequently reared within normal conditions to an average size of 52 g. The long-term hyperthermic embryos showed up-regulated ANP transcription at the ∼9th and ∼20th somite stage and at the completion of somitogenesis. The cardiosomatic index [CSI; (ventricle weight/body weight) ∗100] demonstrated a significant decrease in the relative heart weight of fish incubated at 10 °C during the embryogenesis compared with controls. In these fish, aplasia of septum transversum was observed in 2 of 25 fish, resulting in abnormally shaped hearts situated partly within the abdominal cavity. Altogether, our results demonstrate that hyperthermia both induce deviant development of heart and associated structures and up-regulation of ANP transcription during embryogenesis. A possible role of ANP in development of heart malformations is thus suggested.