AMPK alpha subunit gene characterization in Artemia and expression during development and in response to stress

Abstract

AMP-activated protein kinase (AMPK) plays a central role in maintaining the energy balance of organisms under physiological and environmental stresses. Here two AMPK alpha subunit gene transcripts (named Afr-AMPKalpha1 and Afr-AMPKalpha2) from Artemia franciscana were isolated and gene expression was characterized by semiquantitive reverse transcription-polymerase chain reaction (RT-PCR). Afr-AMPKalpha1 was differentially expressed during Artemia developmental stages as well as in response to stresses, such as heat-shock, starvation and a hyperosmotic environment. Afr-AMPKalpha1 mRNA expression in adult Artemia decreased under heat shock, but not in a time- and temperature-dependent manner. By contrast, the transcript sharply decreased in heat-shocked cysts in a time-dependent manner. Under hyperosmotic stress, however, the mRNA level in adults first declined and then increased with prolonged exposure. In the case of starvation, the gene expression in adults decreased and was undetectable after day 9. In addition, Afr-AMPKalpha2 mRNA expression was too low to be detected without nested PCR. Southern blot analysis, moreover, indicated AMPK alpha subunit was present in multiple copies in the Artemia genome. Furthermore, our results demonstrate that the Afr-AMPKalpha1 mRNA level sharply decreases in Artemia carrying diapause-destined embryos and this indicates the possibility that Afr-AMPKalpha1 is involved in determining the reproductive mode in Artemia.