NMR-based untargeted metabolomic study of hydrogen peroxide-induced development and diapause termination in brine shrimp

Artemia diapause has been extensively studied in embryonic biology for a long time. It has been demonstrated that hydrogen peroxide (H2O2) can increase the hatching rate in the development and diapause termination of Artemia cysts. This study used an untargeted 1H NMR-based metabolomic approach to explore the physiological regulation of H2O2 in initiating the development and terminating the diapause of Artemia cysts. This experiment was divided into two parts. In the first part, we analyzed three groups with or without H2O2 as control-0 h, control-5 h and H2O2 (180 μM)-5 h; in the second part, after 7-d incubation, the non-hatching cysts were treated with different H2O2 concentrations as low as 180 μM and as high 1800 μM. The results showed that arginine and proline metabolism were up-regulated after 5 h, and H2O2 up-regulated valine, leucine and isoleucine biosynthesis in the development of cysts. In the second part, low H2O2 (180 μM) showed alanine, aspartate and glutamate metabolism, but high H2O2 (1800 μM) also up-regulated arginine and proline metabolism, as in the control group without H2O2 stimulus. These results suggest that enough H2O2 can catalyze cell transcription and translation in Artemia cysts, and it improves the cell growth rate, thus allowing embryo cells to grow again.