Molecular characterization and expression of sirtuin 2, sirtuin 3, and sirtuin 5 in the Wuchang bream (Megalobrama amblycephala) in response to acute temperature and ammonia nitrogen stress

Abstract

This study sought to characterize sirtuin 2 (sirt2), sirtuin 3 (sirt3), and sirtuin 5 (sirt5) in Megalobrama amblycephala (M. amblycephala) by cloning the open reading frame (ORF) of sirt2, sirt3, and sirt5. The full-lengths of the resulting M. amblycephala sirt2, sirt3, and sirt5 cDNA sequences were 1845, 1534, and 1920 bp, respectively, with 92%, 98%, and 91% similarities to Danio rerio sequences. Based on our bioinformatic analyses and predictions, the sirt2 and sirt3 genes of M. amblycephala were classified within the Sir2 I family, whereas sirt5 belonged to the Sir2 III family. Furthermore, sirt2, sirt3, and sirt5 were widely distributed in different M. amblycephala tissues. Particularly, sirt2 and sirt5 were highly expressed in gills, intestines, and liver (P < 0.05), whereas sirt3 was highly expressed in gills, kidney, liver, and spleen (P < 0.05). A 2 × 2 factorial experiment was also conducted to analyze sirt2, sirt3, and sirt5 expression patterns in response to acute temperature (25 and 32 °C) and ammonia nitrogen (0 and 20 mg/L) stress. Notably, these two stressors were found to interactively affect sirt2, sirt3, and sirt5 expression patterns in M. amblycephala liver. At the higher water temperature (32 °C) and ammonia nitrogen concentration (20 mg/L) tested herein, sirt2, sirt3, and sirt5 had similar expression levels and exhibited a down-regulation trend at 6 and 48 h post-stress but became up-regulated thereafter to counteract the stressors at 96 h post-stress.

Keywords: Ammonia nitrogen; Megalobrama amblycephala; Molecular cloning; Sirtuin family; Temperature.