System for evaluation of oxidative stress on in-vitro-produced bovine embryos

• Published in: Reproductive biomedicine online 2015-10, Vol.31 (4), p.577-580
• Main Authors: de Assis, Patricia Monken, Castro, Leticia Signori, Siqueira, Adriano Felipe Perez, Delgado, Juliana de Carvalho, Hamilton, Thais Rose dos Santos, Goissis, Marcelo Demarchi, Mendes, Camilla Mota, Nichi, Marcilio, Visintin, José Antonio, Assumpção, Mayra Elena Ortiz D'Ávila
• Format: Article
• Language: English
• Subjects: Animals; Blastocyst - cytology; Blastocyst - drug effects; Blastocyst - metabolism; Blastomeres - cytology; Blastomeres - drug effects; Blastomeres - metabolism; Cattle; CellROX®Green; development; Embryo Culture Techniques; embryo oxidative stress; Embryo, Mammalian - cytology; Embryo, Mammalian - drug effects; Embryo, Mammalian - metabolism; Embryonic Development - drug effects; Embryonic Development - physiology; Female; Fertilization in Vitro - veterinary; microscopy of epifluorescence; Microscopy, Fluorescence; Obstetrics and Gynecology; Oxidative Stress - drug effects; Vitamin K 3 - toxicity
• ISSN: 1472-6483; 1472-6491
• DOI: 10.1016/j.rbmo.2015.06.014

Abstract This study proposed a quantitative evaluation of oxidative status (OS) in bovine embryos. Sixteen-cell stage embryos, cultured under 5% O2 , were treated with oxidative stress inducer menadione (0, 1, 2.5 and 5 µmol/l) for 24 h. Blastocyst rate (BLR) was recorded and expanded blastocysts were stained with CellROX®Green (CRG; OS evaluation) and evaluated under epifluorescence microscopy (ratio of pixel/blastomere). A significant effect of menadione was observed for BLR ( P = 0.0039), number of blastomeres/embryo ( P < 0.0001) and OS ( P < 0.001). Strong negative correlations were found between BLR and the number of blastomeres with OS evaluation, demonstrating the efficacy of this analysis to evaluate OS levels of IVF bovine embryos.