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The objective of the study was to histologically describe the structure of the testis and epididymis, identifying the GAGs in them. Furthermore, the distribution of smooth muscle alpha‐actin, desmin, and vimentin is described of E. bilineatus, E. perditus, and U. vautieri, through immunohistochemical method. Two adult males of each species of the lizards were collected in southeastern Brazil. In the testicular albuginea, the presence of neutral glycoproteins was detected. In E. bilineatus, the epididymal duct epithelium is simple, composed of secretory cells, cylindrical in shape and acidophilic cytoplasm. In the testis of the three lizards there was a positive immunolocation accentuated for α‐SMA in the dense connective tissue of the tunica albuginea. A moderate reaction was observed in the connective tissue albuginous and in the layer of smooth muscle fibers in a circumferential arrangement of the epididymal duct. The immunolocation for desmin was accentuated in the fibers of the testicular albuginea of E. bilineatus, while in U. vautier and E. perditus was moderate. In the epididymis, only in E. perditus and E. bilineatus positive immunoreaction was verified. In the three lizards there were no observations of immunostaining for vimentin with the antibody utilized.