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Disruption of the endothelial nitric oxide synthase gene affects ovulation, fertilization and early embryo survival in a knockout mouse model

Fundacion Centro Nacional de Investigaciones Cardiovasculares Carlos III (CNIC), Melchor Fernandez Almagro 3, 28029 Madrid, Spain¹ Dpto. de Medicina y Cirugia Animal, Facultad de Veterinaria, UCM, 28040 Madrid, Spain² Dpto. de Reproduccion Animal, INIA, Avda. Puerta de Hierro s/n. 28040 Madrid, Spain³ Instituto de Ciencia Animal y Tecnologia de Carnes, UACh, Casilla 567, Valdivia, Chile

Correspondence should be addressed to A Gonzalez-Bulnes; Email: bulnes{at}inia.es

Two consecutive experiments determined whether disruption of the endothelial nitric oxide synthases (NOS) gene (Nos3) affects ovulation, fertilization, implantation, and embryo development. In the first trial, Nos3-knockout mice (groups Nos3^–/–) and wild-type mice (groups Nos3^+/+) showed significant differences in mean number of corpora lutea (9.7±1.2 in Nos3^–/– versus 14.2±1.2 in Nos3^+/+; P<0.01), rate of anovulation (48.3±7.3% in Nos3^–/– versus 29.7±6.3 in Nos3^+/+; P<0.05), total mean number of recovered oocytes/zygotes (4.0±1.1 in Nos3^–/– versus 10.4±1.6 in Nos3^+/+; P<0.01), and non-fertilization rate (50.7 in Nos3^–/– versus 3.3% in Nos3^+/+; P<0.001). In the second trial, implantation and early pregnancy losses in Nos3-knockout and wild-type dams were detected by real-time ultrasound imaging. The number of embryos reaching implantation was higher in Nos3^+/+ than in Nos3^–/– mice (7.5±0.4 vs 4.0±0.4; P<0.005); thereafter, embryo losses were detected between days 8.5 and 13.5, in 62.5% of the Nos3-knockout dams and, at days 10.5 and 11.5, in 16.7% of the control females (P<0.005). Thus, NO and NOS3 deficiencies affect reproductive and developmental features in the Nos3-knockout mouse model.