This prospective study compares the diagnostic and predictive value of MND1 gene expression in testicular dysfunction in rats. Thirteen rats received a single intraperitoneal injection of 10 mg/kg of body weight busulfan to induce azoospermia, and 15 untreated rats remained as the control group. After 35 days, blood and testis were collected for analysis. MND1 gene expression was done by PCR. Follicle-stimulating hormone (FSH), luteinizing hormone (LH) and testosterone were determined in blood by ELISA. Oxidative stress markers and lipid profile were determined colimetrically. Histopathological and immunohistochemical studies were used H&E and YAP stain. Body weight, MND1 gene expression, TAC and testosterone were decreased significantly in busulfan-treated rats compared to control (p<0.0001). Serum FSH, LH, NO, LPO, and HDL were significantly increased in busulfan treated group compared to control.  Testis of busulfan-treated rats showed a significant decrease in YAP immuno-expression compared to control. In conclusion, significant decrease in MND1 expression in testicular tissue with increase in serum level of FSH and LH are promising predictor for testicular function.