We present first data on reproductive biology of male blackspotted smooth-houndMustelus punctulatusin the Adriatic Sea and first histological examination of gonads at different maturation stages and seasonal cycles for this species. We collected samples from 117 male specimens ofM. punctulatusranging in length from 44.6 to 126.5 cm, caught by commercial bottom trawls in the North-Central Adriatic Sea. Microscopic observation revealed a diametric development of testes, in which round-shaped spermatocysts have zonal arrangement, showing seven different stages of development. Males began to mature when they were between 61 and 88 cm body length, which was indicated by the presence of functional claspers or appearance of the spermatogenic cysts. While the smallest mature shark was only 80 cm long, all animals longer than 89 cm were sexually mature. The seasonal analysis of the testes indicated that mature spermatozoa dominated in testes during the first half of the year with a peak in May and June, after which followed less active period during July and August.