Pfaffia glomerata polyploid accession compromises male fertility and fetal development.

ETHNOPHARMACOLOGICAL RELEVANCE: Pfaffia glomerata (Spreng.) Pedersen has traditionally been used as a tonic and a stimulant by the Brazilian population. It shows higher biomass accumulation and production of secondary compounds, such as the phytosterol 20-hydroxyecdysone. AIMS: The present study aimed to evaluate the effects of the hydroalcoholic extract of the root of tetraploid P. glomerata (BGEt) on testicular parenchyma, and its implications on fertility. MATERIAL AND METHODS: Adult Swiss mice were divided as: control (water) and sildenafil citrate (7 mg/kg), BGEt at 100, 200, and 400 mg/kg, and BGEtD 200 mg/kg (treated with BGE every three days). Males (n = 4/group) were mated with normal untreated adult females to assess fertility rates, while other animals (n = 6/group) were euthanized for testis, epididymis, and oxidative stress analyses. RESULTS: Increase in tubule diameter and epithelium height in the discontinuous group, in addition to an increase in the proportion of tubules with moderate pathologies was observed. The pre-implantation loss was lower in all treated groups. The post-implantation loss was significantly increased in all treated groups, except for the lowest BGEt dose. BGEt intake caused a decrease in daily sperm production, along with the number and quality of sperm in the epididymis. Changes were observed in protein carbonylation and hydrogen peroxide and nitric oxide levels, characterizing oxidative stress. CONCLUSIONS: The hydroalcoholic extract of P. glomerata tetraploid altered sperm and testicular parameters, compromising embryonic development after implantation.

Alterations in the testicular parenchyma of Foxn1+/- and Foxn1-/- adult mice.

Nude mice carry an autosomal recessive mutation in the Foxn1 gene and therefore are homozygous recessive animals (Foxn1 -/-). The fertility rate of homozygous male (Foxn1-/- ) is low, which seems to be related to the delay in the production of gametes at the beginning of sexual maturity. The present study evaluated the structural and organizational aspects of the testicles of homozygous and heterozygous offspring related to the Foxn1 gene in mice, describing its implications on spermatogenesis. Adult males Balb/c, Foxn1+/- and Foxn1-/- mice were used. Testes and epididymis were harvested for histological, biochemical, and sperm transit analyses. Gonadal weight was significantly lower in Foxn1+/- and Foxn1-/- animals, the same behavior was noticed for the activity of antioxidant enzymes. In addition, tubular parameters such as epithelial proportion, length, and area, as well as germ and Leydig cell's populations were significantly reduced in the aforementioned groups, leading to lower sperm production. In conclusion, our results indicate the importance of the Foxn1 in Leydig cell's function, reflecting in the preservation of spermatogenesis, thus in germ cell's population and sperm cell production.

Effects of Dietary Crude Glycerin Concentration on Testicular Morphology and Oxidative Stress Markers and on Plasma Testosterone Concentrations.

We evaluated the effects of feeding 6%, 12% or 18% crude glycerin, containing 80.5% glycerol, on testicular histomorphometry and markers of oxidative stress and on plasma testosterone concentrations in lambs. Body weight, testicular biometric measurements, gonadosomatic index and net weight of the testicles were higher for the treated groups (P <0.05) compared with a control group that did not receive dietary glycerin. The mean total length of seminiferous tubules was higher in the 6% group (P <0.05), while the mean total tubular and seminiferous epithelium volumes increased in all treated groups (P <0.05). The volume of Leydig cells increased in the 12% group, while their number per gram of testicle decreased (P <0.05). There was a decrease in mean nuclear diameter and mean volume of Leydig cells, and an increase in the mean number of these cells per gram of testicle, in the 18% group (P <0.05). Plasma testosterone concentrations were unaffected. There was desquamation of seminiferous epithelium and vacuolation of Sertoli cells in the treated groups. Variable degrees of spermatocyte necrosis and the presence of giant cells were seen in all groups and there was intense vacuolation of Sertoli cells in the 12% and 18% groups. Superoxide dismutase and catalase production increased most in the 12% and 18% groups (P <0.05), while glutathione production was higher in the 18% group (P <0.05). Mean nitric oxide concentration decreased in all treated groups (P <0.05), while malondialdehyde production was higher in the 18% group than in the control and 6% groups (P <0.05). We conclude that the inclusion of 6% glycerin in the diet of lambs results in changes in testicular morphology that have been previously associated with improved reproductive function, but without evidence of oxidative stress.

Seasonal evaluation of spermatogenesis of the hematophagous bat Desmodus rotundus in the Caatinga biome.

This study was aimed to characterize the spermatogenic process and its seasonal variation in Desmodus rotundus, in the Caatinga biome, a water-limited ecosystem, with marked water restriction during most of the year. Collections of adult animals were performed during the dry and rainy seasons, and after euthanasia, their testes were processed histologically to perform morphological, morphometric, ultrastructural and immunohistochemical analyzes. The percentage of seminiferous epithelium, number of Leydig cells per gram of testis, and population of Sertoli cells and A-type spermatogonia presented by D. rotundus were significantly higher in the rainy season, while the percentage of lumen, mitotic index, support capacity performed by Sertoli cells, and overall yield of spermatogenesis were higher in the dry season. The ultrastructure of spermatogenesis was similar to that described in other mammals, and the immunohistochemical analysis revealed activity of the aromatase enzyme in Sertoli cells, Leydig cells, spermatocytes and spermatids, as well as the presence of androgen receptors in Sertoli cells and Leydig cells. FGF2 activity was detected in primary spermatocytes in zygotene and pachytene, as well as secondary spermatocytes and rounded and elongated spermatids, while the BCL-2 protein was expressed in primary spermatocytes in zygotene and pachytene, secondary spermatocytes, and rounded spermatids. The activity of these molecules was similar in both seasons, and associated with the morphometric findings, indicates maintenance in the integrity of the seminiferous epithelium throughout the year. The seasonal study of D. rotundus spermatogenesis indicates a continuous spermatogenesis pattern and suggests a greater production of spermatozoa in the rainy season in the Caatinga biome.

Spermatogenic dynamics of the spiny rat Kannabateomys amblyonyx (Wagner, 1845) (Rodentia, Echimyidae).

Rodents are distributed worldwide, playing important ecological roles regarding preservation of forest areas. Thus, the study of their reproductive biology is a key for conservation initiatives that prevent extinction and/or improve species management. The present study aimed to describe the spermatogenic dynamics of the spiny rat Kannabateomys amblyonyx, an endemic species from Atlantic Rainforest areas, in Brazil. The average body weight was 418.43g with a gonadosomatic index of 0.41%. The testicular parenchyma organization followed the pattern described for other rodent species, with a large amount of seminiferous tubules occupying 93.57% of the organ, in a total of 26.04m per testicle. Stage I of the seminiferous tubule cycle was the most frequent in K. amblyonyx, while stage IV the most scarce. Each tubular section in stage I showed 0.47 type-A spermatogonia, 11.78 primary spermatocytes in pre-leptotene, 3.81 in zygotene and 14.31 in pachytene, whereas 32.19 cells were round spermatids and 6.23 were Sertoli cells. From these results it was possible to determine the sperm reserve of 274.49×106 cells per gram of testis. The mitotic and meiotic indexes were 25.06 and 2.25 cells, respectively, whereas the spermatogenic yield was 69.73 cells. Those findings are significant since this is the first study regarding the reproductive aspects of the only Echimyidae species in Brazil, which shows a monogamous mating system.