The purpose of this study was to describe the structure of the reproductive tract of males of Potamotrygon magdalenae before, during, and after they acquire sexual maturity, and to establish the first maturity scale for males within the family Potamotrygonidae. The male reproductive tract of P. magdalenae is composed of testes, efferent ducts, epididymides, deferent ducts, seminal vesicles, Leydig, alkaline, and clasper glands, and claspers, all of which are paired and functional. Four sexual maturity stages were established: immature, maturing, reproductively active, and resting. The degree of claspers calcification is also a good indicator of sexual maturity in this species. The testes are lobulated, each lobe contains numerous spermatocysts which are organized in zones and are displaced radially from germinal papillae to the spermatozoa zone where individual spermatozoa are conveyed to the efferent ducts. The epididymis can be regionalized in head, body, and tail; these regions are distinguished by external pigmentation and by the epithelium lining configuration. The tail of the epididymis is connected with the deferent duct and this, in turn, with the seminal vesicle. The spermatozoa are organized in spermatozeugmata which begin to form in the deferent duct; this latter organ is attached laterally at the Leydig gland that is composed by simple glandular units. Irregular and vesicular secretions can be found in the genital ducts. These secretions might be associated with the maturation of the spermatozoa and formation of spermatozeugmata. The male reproductive tract of P. magdalenae is similar to other elasmobranchs; however, two types of primary spermatogonia, an epididymis internally regionalized, and the presence and structure of spermatozeugmata are specific features not yet described in freshwater stingrays. Most of the year, the males were reproductively active, however, few resting adult males occurred during one of the months of the lowest waters.