ABSTRACT. The goal of present study was to evaluate the effects of diabetes on quantitative parameters of Leydig cells. Twelve adults Wistar rats were divided in: 1) Diabetic Group (DG), which was induced by a single intraperitoneal injection of streptozotocin (60 mg kg -1 of body weight); and 2) Control Group (CG), which received citrate buffer intraperitoneal. After eight weeks of diabetic induction, the animals were weighted, anesthetized and testicles were removed and routinely processed to paraffin embedded. Body weight (40%) and testicular weight (18%) of diabetic rats were significantly lower than control group. Diabetic rats showed an increase in interstitial compartment but the tubular compartment did not differ. The individual volume of Leydig cells and nuclear diameter were lower in DG. However, the population of these cells was increased. In conclusion, diabetes induced by streptozotocin in adult rats promoted alterations in testicular compartments and changes on volume, nuclear diameter and population of Leydig cells, compromising the testicular function.Keywords: streptozotocin, testes, interstitial cells, morphometry. Avaliação de parâmetros quantitativos de células de Leydig em ratos diabéticos adultosRESUMO. O objetivo do presente estudo foi avaliar os efeitos do 'diabetes' nos parâmetros quantitativos de células de Leydig. Doze ratos machos adultos foram divididos em: 1) Grupo Diabético (GD) induzidos por injeção intraperitoneal única de estreptozotocina (60 mg kg -1 de peso corporal); e 2) Grupo Controle (GC) receberam tampão citrato, via intraperitoneal. Após oito semanas da indução, os animais foram pesados, anestesiados, os testículos foram removidos e processados rotineiramente em parafina. O peso corporal (40%) e testicular (18%) dos ratos diabéticos reduziu significativamente em relação ao grupo controle. Ratos diabéticos mostraram aumento no compartimento intersticial, mas o compartimento tubular não apresentou diferença significativa. O volume individual e o diâmetro nuclear de células de Leydig reduziram em GD. No entanto, a população dessas células aumentou. Em conclusão, o 'diabetes' induzido por estreptozotocina, em ratos adultos, promoveu alterações nos compartimentos testiculares e mudanças no volume, diâmetro nuclear e população das células de Leydig, comprometendo a função testicular.Palavras-chave: estreptozotocina, testículo, células intersticiais, morfometria.

Type 2 diabetes mellitus (T2D) during pregnancy is characterized by high levels of reactive oxygen species and pro-inflammatory factors in the placenta. Once these reactive species reach the foetus, they trigger physiological adaptations that allow the foetus to survive, but programme the organism to develop metabolic disorders in adulthood. The male reproductive system is highly susceptible to foetal programming. This study aimed to investigate the effects of intrauterine exposure to T2D on testicular histomorphometry and redox homeostasis of adult rats and evaluate the effects of maternal treatment with metformin and pentoxifylline. Female rats were induced to T2D, then treated with metformin and pentoxifylline, or co-treated with both drugs. The females were mated, the male offspring were sacrificed on postnatal day 90, and the testicles were collected for analysis. Metformin protected the tubular compartment, with the maintenance of the Sertoli cell population and daily sperm production. Pentoxifylline attenuated the effects of diabetes on Leydig cells, in addition to stimulating testosterone production and lowering lipid peroxidation. Intrauterine exposure to T2D results in important testicular alterations that compromise gonadal function, and the cotreatment with metformin and pentoxifylline may represent a promising therapy that attenuates these effects by combining the positive influences in both the tubular and interstitial compartments of the testicular parenchyma.

Ganciclovir (GCV) has been implicated in the development of testicular alterations. Exposure on gestational day (GD) 10 in rats induced permanent effects, including focal reduction or absence of germ cells (Sertoli cell-only tubules). Because the timing of exposure can be critical for testicular effects, we exposed rat dams to 300 mg/kg GCV (3 100 mg/kg subcutaneous injections) on GD10, 14 and 19, when germ cells have high rates of migration, proliferation and are mitotically quiescent, respectively. Males exposed to GCV in utero on GD10 and 14 were evaluated for androgenization markers, serum and fecal androgens, and testicular histomorphometry at adulthood. Double-labeling immunofluorescence for DAZL and Ki67 were used to assess gonocytes number and the proliferative activity of germ and somatic cells in fetal testes on GD15 and 20, ie, 24 h after GCV exposure. Adult rats exposed on GD14 showed delayed puberty onset, despite normal androgen levels. Also, there was a 50% reduction in testicular weight and about 30% of seminiferous tubules lacking germ cells. Effects on GD10 animals were less pronounced. In the fetal testis, the number of gonocytes was reduced by 50% in rats exposed on GD14, but normal in GD19 fetuses. GCV also reduced Sertoli cell proliferation immunolabeling in GD19 fetuses and Sertoli cell number in adults. In conclusion, GCV toxicity on germ cells seems to be linked to their proliferation rate and GD14 is a critical window in rats, when GCV exposure causes an acute massive loss of germ cells that persists until adulthood.

The females of yellowtail tetra (Astyanax lacustris), known as the freshwater sardine, are approximately 1.33 times larger than males, and thus, all-female monosex culture would increase production and reduce size variability. The present work aimed to identify the optimal dose of 17α-methyltestosterone (MT) to be used in the masculinization of A. lacustris for indirect sex reversal. Three different concentrations of MT (20, 40, and 60 mg/kg of feed in the diet) were fed to the fry for 30 days. Thirty adult individuals from each treatment, including the control (0 mg MT/kg), were evaluated for gonadal development, morphological and histological sexual identification, zootechnical performance, and the possible genotoxic effect caused by prolonged exposure to MT. MT significantly (P<0.01) affected the differentiation of the gonads, with the presence of possible inhibitory effects in all treatments. Intersex individuals were present in the 20 and 60 mg MT/kg treatments. All treatments were able to masculinize A. lacustris and the treatment with the lowest hormone concentration produced the highest percentage of males 76.7%, while the control had 46.7% males. The presence of erythrocyte nuclear alterations indicated a possible cytotoxic effect of MT in treatments 40 and 60 mg MT/kg, however, the use of the hormone did not affect the growth and the survival of the individuals. Thus, the use of MT is a viable option for obtaining neomales as a first step into the production of all-female progenies.

The aim of this study was to analyze the behavior and histopathological changes in the hippocampus of epileptic Wistar rats treated with acupuncture associated or not with phenobarbital. The experiment used 44 male rats with 90 days of birth, induced to status epileptics with pilocarpine hydrochloride in a single dose of 350mg/kg, separated into treatment groups and submitted for 5 minutes to the elevated plus-maze test. Group 1 received 0.2mL of saline solution orally; Group 2 treated with acupuncture at the yintang, baihui, shishencong, jizhong, naohu, thianzu points; Group 3 received orally phenobarbital, daily dose of 20mg/kg; Group 4 treated with an association of acupuncture and oral phenobarbital; Group 5 random needling. The results obtained showed that Groups 2 (acupuncture) and 4 (acupuncture and phenobarbital) presented decreased anxiety, epileptic seizures, and neuronal death in the CA1, CA3 areas of the hippocampus when compared to animals in groups 1, 3 and 5. It is concluded that the association of phenobarbital and acupuncture points used in the experiment allowed for the control of epileptic seizures, reduction of anxiety and reduction of lesions in the subareas of the hippocampus.