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

Abstract

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-0h, control-5h and H₂O₂ (180μM)-5h; in the second part, after 7-d incubation, the non-hatching cysts were treated with different H₂O₂ concentrations as low as 180μM and as high 1800μM. The results showed that arginine and proline metabolism were up-regulated after 5h, and H₂O₂ up-regulated valine, leucine and isoleucine biosynthesis in the development of cysts. In the second part, low H₂O₂ (180μM) showed alanine, aspartate and glutamate metabolism, but high H₂O₂ (1800μM) 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.