Fig 1.
A total of 48 adult and 48 prepubertal Sprague Dawley rats were divided into four treatment groups. The first group served as a fertile control. The second group underwent unilateral orchiectomy. The third group was treated with busulfan injections. The last group got the combination of a unilateral orchiectomy and busulfan injections. D: day.
Fig 2.
Evolution of testosterone levels of rats treated at adult (a) or prepubertal (b) age. Variables with a different letter are statistically different. FC: fertile controls, O: rats treated with unilateral orchiectomy, B: rats treated with busulfan, O+B: rats treated with busulfan and unilateral orchiectomy, D: day.
Fig 3.
Bodyweight (A) and testicular weight (B) of the adult treated rats at the time of euthanasia. Bodyweight (C) and testicular weight (D) of the prepubertal treated rats at the time of euthanasia. Variables with a different letter are statistically different. If no letters are indicated, no statistical differences were found in the graph. FC: fertile controls, O: rats treated with unilateral orchiectomy, B: rats treated with busulfan, O+B: rats treated with busulfan and unilateral orchiectomy.
Fig 4.
Evaluation of spermatogenesis and number of spermatogonia.
a) A UCHL1 staining was performed to evaluate the number of spermatogonia (white arrow) per tubule. All pictures show testis sections of prepubertal treated rats. Left row: DAPI, middle row: UCHL1, right row: merge UCHL1+DAPI. b) A HE staining was performed to evaluate spermatogenesis. All pictures show testis sections of prepubertal treated rats. Thirty percent of the rats that received the combined treatment at prepubertal age showed SCO tubules. FC: fertile controls, O: rats treated with unilateral orchiectomy, B: rats treated with busulfan, O+B: rats treated with busulfan and unilateral orchiectomy.
Fig 5.
a) Average number of spermatogonia per tubule. Variables with a different letter are statistically different. If no letters are indicated, no statistical differences were found in the graph. b) Proportion of tubules with ongoing spermatogenesis. A: treated at adult age, P: treated at prepubertal age, FC: fertile controls, O: rats treated with unilateral orchiectomy, B: rats treated with busulfan, O+B: rats treated with busulfan and unilateral orchiectomy.
Fig 6.
Number of litters produced during the whole mating experiment per adult (a) or prepubertal (c) treated rat per gestation period during 4 months of mating. Mean number of pups per litter produced per adult (b) or prepubertal (d) treated rat per gestation period. Variables with a different letter are statistically different. If no letters are indicated, no statistical differences were found in the graph. FC: fertile controls, O: rats treated with unilateral orchiectomy, B: rats treated with busulfan, O+B: rats treated with busulfan and unilateral orchiectomy.