Abstract: The aim of this study was to investigate the role of Nitric Oxide (NO) in the atresia of ovarian follicles in an animal model. Twenty adult, female rats (90 days old with body weights of 210 ± 10 g in the beginning of the experiments) were divided into 4 groups of 5 each. They were treated twice daily from the subcutaneous route for 21 successive days with either of the following chemicals: nitroglycerine, L-arginine, L-NAME, or saline. On day 22, all animals were sacrificed. Ovaries were dissected out free of connected tissue and were fixed in formaline 10%. Later, paraffine blocks were prepared and serial sections were made by means of H and E routine staining method. Intact and atretic follicles were counted separately. In addition, damages were analyzed qualitatively from the points of view of appearance and morphologic changes. In the evaluation of ovarian follicular structures, different types of healthy as well as atretic follicles were observed. In most of atretic follicles, the oocytes were abnormally elongated and increnation of their outlines were obvious. There were numerous macrophages around and inside of the atretic follicles. Our investigation regarding the distribution of atretic follicles in the ovaries of test groups revealed that atretic follicles in the L-NAME treated group were increased in comparison to the control group. Conversely, however, in the arginine-treated group, the atretic follicles were reduced compared to the control animals. Treatment with nitroglycerine of the rats decreased the number of atretic follicles significantly (p<0.05) in comparison to the control group. In conclusion, enhanced NO, either from endogenous or exogenous origins, prevents atresia phenomenon, while inhibition of NO exerts an opposite effect.