Nitric oxide modulates mitochondrial activity and apoptosis through protein S-nitrosylation for preimplantation embryo development

• Published in: Journal of assisted reproduction and genetics 2013-08, Vol.30 (8), p.1063-1072
• Main Authors: Lee, Tsung-Hsien, Lee, Maw-Sheng, Huang, Chun-Chia, Tsao, Hui-Mei, Lin, Pi-Mei, Ho, Hong-Nerng, Shew, Jin-Yuh, Yang, Yu-Shih
• Format: Article
• Language: English
• Subjects: Animals; Apoptosis; Blastocyst - cytology; Blastocyst - drug effects; Blastocyst - ultrastructure; Embryo Biology; Embryo Culture Techniques; Embryonic Development - drug effects; Embryos; Endometriosis; Endoplasmic reticulum; Fertility; Gynecology; Human Genetics; Infertility; Medicine; Medicine & Public Health; Mice; Mitochondria; Mitochondria - drug effects; Mitochondria - metabolism; Mitochondria - physiology; Nitric oxide; Nitric Oxide - toxicity; Obstetrics; Oxidative stress; Proteins; Reproductive Medicine
• ISSN: 1058-0468; 1573-7330
• DOI: 10.1007/s10815-013-0045-7

Purpose Previous studies reported that patients with endometriosis had excess nitric oxide (NO) in the reproductive tract and poor embryo development in IVF cycles. This study aims to elucidate the effects of NO on early embryo development. Methods Zygotes from superovulated B6CBF1 mice were cultured to blastocysts in a variety of media. Sodium nitroprusside (SNP) and N G -nitro-L-arginine (LNA) were added to the culture medium as a NO donor and a NO synthase inhibitor, respectively. The localization and fluorescence intensity of S-nitrosylated (SNO) proteins within 2-cell stage embryos were analyzed with confocal microscopy. Apoptosis and ATP production in the blastocysts were measured. Result(s) Subsequent to NO exposure, the SNO proteins mainly colocalized with the mitochondria and endoplasmic reticulum and the intensity of SNO proteins increased. The addition of a quanylate cyclase inhibitor and a cyclic GMP mimic agent induced nonsignificant changes in SNO proteins, whereas addition of a superoxide scavenger or a reduced form of glutathione rescued the embryos from the effects of NO. However, superoxide scavenger supplementation resulted in decreased blastocyst ATP production. Conclusion(s) Elevated NO exerts deleterious effects on embryo development, possibly through protein S-nitrosylation in the mitochondria and endoplasmic reticulum. Including glutathione as a component in the culture medium might counteract this effect.