Chlamydia trachomatis (CT) is one of the most common sexually transmitted pathogens. The consequences of CT has been extensively studied in models of female genital tract infections. On the other hand, male genital tract infections with CT are not well understood, specially prostatic infections, due to the lack of a versatile experimental model. In this work adult male mice (n= 20) were inoculated with CT (1x107 inclusion-forming units) in the meatus urethra. A control group (n=20) was sham infected. Animals were sacrificed at 4, 7, 10, 14, 20 and 30 days postinfection (dpi) and urethra, seminal vesicles, prostate and testis were removed aseptically, homogenized and inoculated onto LLCMK2 cells to reisolate CT. In addition, blood samples were collected at 20 dpi to evaluate the humoral response by indirect immunofluorescent assays. CT were reisolated at 4 and 15 dpi from urethra in 65% of the animals whereas it could be only reisolated from seminal vesicles at 7 dpi in 50% of animals and from testis in 50% and 30% at 7 and 10 dpi, respectively. Interestingly, CT were reisolated from prostate at 4, 7, 10 and 14 dpi in 35%, 100%, 65% and 35% of the animals repectively. At 20 and 30 dpi all tissues showed negative cultures. Chlamydia-specific IgG were detected 20 dpi with titers higher than 1/400. Animals from control group did not show positive reisolation or CT-specific IgG. These findings suggest that CT could infect different tissues from the genital tract being the prostate highly susceptible.