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Genetic divergence and environment influence on speciation process are the great deal studies over recent decades. One of the best ways for exploring the interaction of geography and genetics is the evaluation of hybrids in a contact zone. To understand if there is one or more hybrid zone between house mouse subspecies in Iran and what are the differences comparing these zones with European well-known hybrid zone, we performed this approach. Samples were live-trapped from Ilam city in west for sensu lato M. m. domesticus subspecies, and Neishabur city in north-east of Iran for sensu lato M. m. musculus subspecies. In five experimental groups, male and female mice of the two subspecies were crossed reciprocally to generate F1 hybrids, and then F1 offspring males and females were crossed also reciprocally between siblings to generate F2 hybrids. In the same manner as seen in European hybrid zone, hybridization between female M. m. musculus and male M. m. domesticus of all five groups showed male sterility in F1 generation, but intact female offspring. These sterile males comparing with a parent or healthy males showed low count and more abnormal sperm percentage in morphological and testis histological section studies. Comparing the results from this study with numerous studies carried out during several years on the European hybrid zone showed an equal condition of contact between two subspecies. Genetical elements have kept their same influence on postzygotic reproductive isolation more than environmental effects far from the Europe, here in Iran.