RESUMO
Several plant species such as Pfaffia glomerata are widely used in traditional Brazilian medicine as stimulants and aphrodisiacs. In this regard, the aim of our study was to explore the effects of the long-term intake of the hydro-alcoholic root extract of P glomerata on the germ and somatic cells within the seminiferous tubules in adult Balb/c mice. The experimental groups were placed as: controls (water and DMSO), and treated with 300 and 400 mg/kg of the root extract. The number of germ and somatic cells, the proportion of pathological seminiferous tubules, and the germ cell apoptotic levels were evaluated. The volume and proportion of the seminiferous epithelium was decreased after the extract intake due to the increased germ cell apoptotic levels. Vacuolization of Sertoli cell cytoplasm was observed widely in pathological tubules, along with fully disorganized epithelia, showing multinucleated cells, which lead to decreased daily sperm production. Taken together, our results indicate that long-term intake of the P glomerata caused deleterious effects on spermatogenesis by inducing apoptosis and altering the seminiferous tubule's epithelial dynamics.
Assuntos
Amaranthaceae/química , Extratos Vegetais/farmacologia , Epitélio Seminífero/efeitos dos fármacos , Espermatogênese/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Células Germinativas/efeitos dos fármacos , Células Germinativas/patologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Raízes de Plantas/química , Epitélio Seminífero/patologia , Túbulos Seminíferos/efeitos dos fármacos , Túbulos Seminíferos/patologia , Células de Sertoli/efeitos dos fármacos , Células de Sertoli/patologiaRESUMO
The existence of cytoplasmic passages between germ cells and their potential function in the control of the spermatogenic process has long been an intriguing question. Evidence of the important role of such structures, known as intercellular bridges (ICB), in spermatogenesis has been implicated by the failure of spermatogenesis in testis-expressed gene 14 (Tex14) mutant mice, which lack the ICBs, to progress past the pachytene spermatocyte stage. Using these Tex14 mutants, the present study evaluated, for the first time, the behavior and synchrony of the spermatogonial lineage in the absence of ICBs. Our data suggest that the absence of these cytoplasmic connections between cells affects the expansion of the undifferentiated type A (Aundiff) spermatogonia compartment and their transition to A1, resulting in a significant numerical reduction of differentiating A1 spermatogonia, but did not interfere with cell amplification during subsequent mitotic steps of differentiating spermatogonia from A1 through intermediate (In). However, beginning at the type B spermatogonia, the synchrony of differentiation was impaired as some cells showed delayed differentiation compared to their behavior in a normal seminiferous epithelium cycle. Thus although spermatogonial development is able to proceed, in the absence of ICBs in Tex14-/- mutants, the yield of cells, specific steps of differentiation, the synchrony of the cell kinetics, and the subsequent progression in meiosis are quantitatively lower than normal.
Assuntos
Comunicação Celular , Diferenciação Celular , Meiose , Epitélio Seminífero/patologia , Espermatogênese , Espermatogônias/patologia , Fatores de Transcrição/fisiologia , Animais , Proliferação de Células , Citoplasma , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Epitélio Seminífero/metabolismo , Espermatogônias/metabolismoRESUMO
One of the most consumed pesticides in the world is glyphosate, the active ingredient in the herbicide ROUNDUP®. Studies demonstrate that glyphosate can act as an endocrine disruptor and that exposure to this substance at critical periods in the developmental period may program the fetus to induce reproductive damage in adulthood. Our hypothesis is that maternal exposure to glyphosate during pregnancy and lactation in mice will affect the development of male reproductive organs, impairing male fertility during adult life. Female mice consumed 0.5% glyphosate-ROUNDUP® in their drinking water [glyphosate-based herbicide (GBH) group] or filtered water [control (CTRL) group] from the fourth day of pregnancy until the end of the lactation period. Male F1 offspring were designated, according to their mother's treatment, as CTRL-F1 and GBH-F1. Female mice that drank glyphosate displayed reduced body weight (BW) gain during gestation, but no alterations in litter size. Although GBH male F1 offspring did not exhibit modifications in BW, they demonstrated delayed testicular descent. Furthermore, at PND150, GBH-F1 mice presented a lower number of spermatozoa in the cauda epididymis and reduced epithelial height of the seminiferous epithelium. Notably, intratesticular testosterone concentrations were enhanced in GBH-F1 mice; we show that it is an effect associated with increased plasma and pituitary concentrations of luteinizing hormone. Therefore, data indicate that maternal exposure to glyphosate-ROUNDUP® during pregnancy and lactation may lead to decreased spermatogenesis and disruptions in hypothalamus-pituitary-testicular axis regulation in F1 offspring.
Assuntos
Glicina/análogos & derivados , Herbicidas/toxicidade , Exposição Materna/efeitos adversos , Efeitos Tardios da Exposição Pré-Natal/induzido quimicamente , Espermatogênese/efeitos dos fármacos , Animais , Animais Lactentes , Modelos Animais de Doenças , Feminino , Ganho de Peso na Gestação/efeitos dos fármacos , Glicina/toxicidade , Humanos , Sistema Hipotálamo-Hipofisário/efeitos dos fármacos , Sistema Hipotálamo-Hipofisário/metabolismo , Lactação , Hormônio Luteinizante/sangue , Hormônio Luteinizante/metabolismo , Masculino , Camundongos , Gravidez , Efeitos Tardios da Exposição Pré-Natal/sangue , Efeitos Tardios da Exposição Pré-Natal/patologia , Epitélio Seminífero/efeitos dos fármacos , Epitélio Seminífero/patologia , Contagem de Espermatozoides , Espermatozoides/efeitos dos fármacos , Espermatozoides/crescimento & desenvolvimento , Testosterona/análise , Testosterona/metabolismo , GlifosatoRESUMO
STUDY QUESTION: Can all types of testicular germ cells be accurately identified by microscopy techniques and unambiguously distributed in stages of the human seminiferous epithelium cycle (SEC)? SUMMARY ANSWER: By using a high-resolution light microscopy (HRLM) method, which enables an improved visualization of germ cell morphological features, we identified all testicular germ cells in the seminiferous epithelium and precisely grouped them in six well-delimitated SEC stages, thus providing a reliable reference source for staging in man. WHAT IS ALREADY KNOWN: Morphological characterization of germ cells in human has been done decades ago with the use of conventional histological methods (formaldehyde-based fixative -Zenker-formal- and paraffin embedding). These early studies proposed a classification of the SEC in six stages. However, the use of stages as baseline for morphofunctional evaluations of testicular parenchyma has been difficult because of incomplete morphological identification of germ cells and their random distribution in the human SEC. STUDY DESIGN, SIZE, DURATION: Testicular tissue from adult and elderly donors with normal spermatogenesis according to Levin's, Johnsen's and Bergmann's scores were used to evaluate germ cell morphology and validate their distribution and frequency in stages throughout human spermatogenesis. PARTICIPANTS/MATERIALS, SETTING, METHODS: Testicular tissue from patients diagnosed with congenital bilateral agenesis of vas deferens (n = 3 adults) or prostate cancer (n = 3 elderly) were fixed in glutaraldehyde and embedded in araldite epoxy resin. Morphological analyses were performed by both light and transmission electron microscopy. MAIN RESULTS AND THE ROLE OF CHANCE: HRLM method enabled a reliable morphological identification of all germ cells (spermatogonia, spermatocytes and spermatids) based on high-resolution aspects of euchromatin, heterochromatin and nucleolus. Moreover, acrosomal development of spermatids was clearly revealed. Altogether, our data redefined the limits of each stage leading to a more reliable determination of the SEC in man. LIMITATIONS, REASONS FOR CAUTION: Occasionally, germ cells can be absent in some tubular sections. In this situation, it has to be taken into account the germ cell association proposed in the present study to classify the stages. WIDER IMPLICATIONS OF THE FINDINGS: Our findings bring a new focus on the morphology and development of germ cells during the SEC in human. Application of HRLM may be a valuable tool for research studies and clinical andrology helping to understand some testicular diseases and infertility conditions which remain unsolved. STUDY FUNDING/COMPETING INTEREST: Experiments were partially supported by Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Fundação de Amparo à Pesquisa de Minas Gerais (FAPEMIG) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq). The authors declare that there are no conflicts of interest. TRIAL REGISTRATION NUMBER: Not applicable.
Assuntos
Envelhecimento , Modelos Biológicos , Epitélio Seminífero/ultraestrutura , Espermatogênese , Espermatozoides/ultraestrutura , Adulto , Idoso , Idoso de 80 Anos ou mais , Biópsia , Disgenesia Gonadal/patologia , Humanos , Processamento de Imagem Assistida por Computador , Masculino , Microscopia , Microscopia Eletrônica de Transmissão , Orquiectomia , Tecido Parenquimatoso/citologia , Tecido Parenquimatoso/crescimento & desenvolvimento , Tecido Parenquimatoso/patologia , Tecido Parenquimatoso/ultraestrutura , Neoplasias da Próstata/patologia , Neoplasias da Próstata/cirurgia , Epitélio Seminífero/citologia , Epitélio Seminífero/crescimento & desenvolvimento , Epitélio Seminífero/patologia , Espermatozoides/citologia , Espermatozoides/crescimento & desenvolvimento , Espermatozoides/patologia , Testículo/anormalidades , Ducto Deferente/anormalidadesRESUMO
Several different strategies have been adopted in attempt to recover from chemotherapy-damaged spermatogenesis that is often seen in oncologic patients. In this study, we have evaluated the impact of short period of exposure to busulphan on the haemogram and seminiferous epithelium of adult rats, focusing on spermatogonial depletion and Sertoli cell (SC) integrity. We then examined whether vitamin B12 supplementation improves the haematological parameters and spermatogonia number. The animals received 10 mg/kg of busulphan (BuG) or busulfan+vitamin B12 (Bu/B12 G) on the first and fourth days of treatment. In H.E.-stained testicular sections, the areas of the seminiferous tubule (ST) and seminiferous epithelium were measured. The number of spermatogonia in H.E-stained and PCNA-immunolabelled testicular sections was quantified. The frequency of tubules with abnormal SC nuclei or TUNEL-positive SC was evaluated. Vimentin immunofluorescence in ST was also evaluated. In BuG and Bu/B12 G, the animals showed leukopenia and thrombocytopenia, but the body weight reduced only in BuG. The areas of ST and seminiferous epithelium decreased in Bu/B12 G and BuG. In BuG, the number of H.E.-stained and PCNA-immunolabelled spermatogonia reduced significantly. The frequency of tubules containing abnormal SC nuclei and TUNEL-positive SC increased and the vimentin immunoexpression pattern changed. In Bu/B12 G, the number of H.E.-stained or PCNA-immunolabelled spermatogonia increased fourfold in comparison with BuG. The structural changes in ST after 6 days of busulphan exposure may be associated with the potential effect of this anti-neoplastic agent on SC. The increased number of spermatogonia in the busulphan-treated animals receiving vitamin B12 indicates that this vitamin can be an adjuvant therapy to improve the fertility in male cancer patients.
Assuntos
Células-Tronco Germinativas Adultas/efeitos dos fármacos , Antineoplásicos Alquilantes/toxicidade , Bussulfano/toxicidade , Epitélio Seminífero/efeitos dos fármacos , Vitamina B 12/farmacologia , Células-Tronco Germinativas Adultas/patologia , Animais , Peso Corporal/efeitos dos fármacos , Leucopenia/induzido quimicamente , Leucopenia/prevenção & controle , Masculino , Tamanho do Órgão/efeitos dos fármacos , Ratos Sprague-Dawley , Epitélio Seminífero/patologia , Células de Sertoli/efeitos dos fármacos , Células de Sertoli/patologia , Espermatogênese/efeitos dos fármacos , Testículo/efeitos dos fármacos , Testículo/metabolismo , Testículo/patologia , Trombocitopenia/induzido quimicamente , Trombocitopenia/prevenção & controle , Vimentina/metabolismoRESUMO
Orchiopexy is performed as part of cryptorchidism and testicular torsion treatment. The inflammation caused by the needle and suture penetration has been suggested to be one of the possible causes of subfertility after parenchymal transfixation of the testicles. The purpose of the present study was to investigate testicular alterations after parenchymal transfixation sutures at different ages in rats. Prepubertal, pubertal, and adult rats were submitted to parenchymal suturing (without tying the knots, thus avoiding local ischemic injury) of the right testicle, which was maintained for 4 hours. All animals were subjected to euthanasia on completion of 14 weeks of life. The right testicles were studied as the sutured testicles, whereas the left organs were studied as contralateral. One age-matched control group of rats that was not submitted to any procedure was used for comparison. During euthanasia, sperm were collected from the tail of the epididymal and evaluated for concentration, motility, and viability. Samples from testicular tissue were collected for morphologic analysis. Sperm analysis indicated that only the adult operated animals presented reductions in motility (38.2% of adult vs. 54.1% of control; P = 0.02) and viability (16.6% of adult vs. 24.6% of control; P = 0.003). Several morphologic alterations were noted both in sutured and in contralateral testes at all ages. For instance, the seminiferous epithelium volumetric density of right testicles was reduced from 50.4% in controls to 32.3% in prepubertal operated animals, 45.3% in pubertal operated animals, and 39.4% in adult operated animals (P < 0.05). The seminiferous epithelium volumetric density was also reduced to 39.9% and 39.0% in contralateral testicles of animals operated before and after puberty, respectively (P < 0.05). The animals operated on before puberty and in adulthood showed more testicular morphologic alterations, as seminiferous tubule volumetric density, seminiferous tubule length, and tubular diameter were reduced only in prepubertal and/or adult operated animals. Testicular transfixation in rats led to important morphologic modifications in the ipsilateral and contralateral organs. These alterations were observed regardless of the age when surgery was performed, but they were milder in animals operated on during puberty. Orchiopexy techniques that do not involve the application of testicular transfixation sutures should be recommended.
Assuntos
Orquidopexia/efeitos adversos , Túbulos Seminíferos/patologia , Maturidade Sexual , Espermatozoides/fisiologia , Fatores Etários , Animais , Sobrevivência Celular , Infertilidade Masculina/etiologia , Infertilidade Masculina/patologia , Masculino , Orquidopexia/métodos , Ratos , Ratos Wistar , Epitélio Seminífero/patologia , Contagem de Espermatozoides , Motilidade dos Espermatozoides , Técnicas de Sutura/efeitos adversos , Testículo/patologiaRESUMO
BACKGROUND: Testis-expressed sequence 101 (TEX101) was found to be highly expressed in testis and involved in acrosome reaction in previous studies. Recently, the metastasis suppressor function of TEX101 in cancer was disclosed, but the comprehensive investigation of its expression has rarely been reported. In this study, the expression features of TEX101 in normal human organs and seminoma were systematically analyzed. RESULTS: Immunohistochemistry demonstrated intense staining of TEX101 in human testis tissues; however, its expression in 27 other types of normal human organs, including the ovary, was negligible. Higher expression of TEX101 was observed in the spermatocytes and spermatids of the testis, but relatively lower staining was detected in spermatogonia. Western blotting showed a single TEX101 band of 38 kDa in human testis, but it did not correspond to the predicted molecular weight of its mature form at 21 KDa. Furthermore, we examined seminoma tissues by immunohistochemistry and found that none of the 36 samples expressed TEX101. CONCLUSIONS: Our data confirmed TEX101 to be a testis protein that could be related to the maturation process of male germ cells. The lack of TEX101 in seminoma indicated its potential role in tumor progression. This characteristic expression of TEX101 could provide a valuable reference for understanding its biological functions.
Assuntos
Proteínas de Membrana/metabolismo , Epitélio Seminífero/metabolismo , Seminoma/metabolismo , Neoplasias Testiculares/metabolismo , Western Blotting , Diferenciação Celular , Epitélio/metabolismo , Feminino , Trato Gastrointestinal/metabolismo , Humanos , Imuno-Histoquímica , Tecido Linfoide/metabolismo , Masculino , Tecido Nervoso/metabolismo , Especificidade de Órgãos/fisiologia , Ovário/metabolismo , Epitélio Seminífero/patologia , Seminoma/patologia , Maturação do Esperma/fisiologia , Espermatozoides/crescimento & desenvolvimento , Neoplasias Testiculares/patologia , Testículo/metabolismo , Testículo/patologiaRESUMO
BACKGROUND: Testis-expressed sequence 101 (TEX101) was found to be highly expressed in testis and involved in acrosome reaction in previous studies. Recently, the metastasis suppressor function of TEX101 in cancer was disclosed, but the comprehensive investigation of its expression has rarely been reported. In this study, the expression features of TEX101 in normal human organs and seminoma were systematically analyzed. RESULTS: Immunohistochemistry demonstrated intense staining of TEX101 in human testis tissues; however, its expression in 27 other types of normal human organs, including the ovary, was negligible. Higher expression of TEX101 was observed in the spermatocytes and spermatids of the testis, but relatively lower staining was detected in spermatogonia. Western blotting showed a single TEX101 band of 38 kDa in human testis, but it did not correspond to the predicted molecular weight of its mature form at 21 KDa. Furthermore, we examined seminoma tissues by immunohistochemistry and found that none of the 36 samples expressed TEX101. CONCLUSIONS: Our data confirmed TEX101 to be a testis protein that could be related to the maturation process of male germ cells. The lack of TEX101 in seminoma indicated its potential role in tumor progression. This characteristic expression of TEX101 could provide a valuable reference for understanding its biological functions.
Assuntos
Humanos , Masculino , Feminino , Epitélio Seminífero/metabolismo , Neoplasias Testiculares/metabolismo , Seminoma/metabolismo , Proteínas de Membrana/metabolismo , Especificidade de Órgãos/fisiologia , Ovário/metabolismo , Epitélio Seminífero/patologia , Maturação do Esperma/fisiologia , Espermatozoides/crescimento & desenvolvimento , Neoplasias Testiculares/patologia , Testículo/metabolismo , Testículo/patologia , Imuno-Histoquímica , Diferenciação Celular , Western Blotting , Seminoma/patologia , Trato Gastrointestinal/metabolismo , Epitélio/metabolismo , Tecido Linfoide/metabolismo , Tecido Nervoso/metabolismoRESUMO
BACKGROUND: Doxorubicin is a potent chemotherapeutic drug used against a variety of cancers. It acts through interaction with polymerases and topoisomerase II and free radical production. Doxorubicin activity is not specific to cancer cells and can also damage healthy cells, especially those undergoing rapid proliferation, such as spermatogonia. In previous studies our group showed that etoposide, another topoisomarese II poison, causes irreversible damage to Sertoli cells. Thus, the aim of this study was to address the effects of doxorubicin on Sertoli cell morphology and function and on the seminiferous epithelium cycle when administered to prepubertal rats. METHODS: Prepubertal rats received the dose of 5 mg/Kg of doxorubicin, which was fractioned in two doses: 3 mg/Kg at 15dpp and 2 mg/Kg at 22 dpp. The testes were collected at 40, 64 and 127 dpp, fixed in Bouin's liquid and submitted to transferrin immunolabeling for Sertoli cell function analysis. Sertoli cell morphology and the frequency of the stages of the seminiferous epithelium cycle were analyzed in PAS + H-stained sections. RESULTS: The rats treated with doxorubicin showed reduction of transferrin labeling in the seminiferous epithelium at 40 and 64 dpp, suggesting that Sertoli cell function is altered in these rats. All doxorubicin-treated rats showed sloughing and morphological alterations of Sertoli cells. The frequency of the stages of the seminiferous epithelium cycle was also affected in all doxorubicin-treated rats. CONCLUSIONS AND DISCUSSION: These data show that doxorubicin administration during prepuberty causes functional and morphological late damage to Sertoli cells; such damage is secondary to the germ cell primary injury and contributed to enhance the spermatogenic harm caused by this drug. However, additional studies are required to clarify if there is also a direct effect of doxorubicin on Sertoli cells producing a primary damage on these cells.
Assuntos
Antibióticos Antineoplásicos/toxicidade , Doxorrubicina/toxicidade , Células de Sertoli/efeitos dos fármacos , Maturidade Sexual , Animais , Doxorrubicina/administração & dosagem , Masculino , Ratos , Ratos Wistar , Epitélio Seminífero/química , Epitélio Seminífero/patologia , Células de Sertoli/patologia , Células de Sertoli/fisiologia , Espermatogênese , Testículo/patologia , Transferrina/análiseRESUMO
Sexual differentiation in the brain takes place from late gestation to the early postnatal days. This is dependent on the conversion of circulating testosterone into estradiol by the enzyme aromatase. The glyphosate was shown to alter aromatase activity and decrease serum testosterone concentrations. Thus, the aim of this study was to investigate the effect of gestational maternal glyphosate exposure (50 mg/kg, NOAEL for reproductive toxicity) on the reproductive development of male offspring. Sixty-day-old male rat offspring were evaluated for sexual behavior and partner preference; serum testosterone concentrations, estradiol, FSH and LH; the mRNA and protein content of LH and FSH; sperm production and the morphology of the seminiferous epithelium; and the weight of the testes, epididymis and seminal vesicles. The growth, the weight and age at puberty of the animals were also recorded to evaluate the effect of the treatment. The most important findings were increases in sexual partner preference scores and the latency time to the first mount; testosterone and estradiol serum concentrations; the mRNA expression and protein content in the pituitary gland and the serum concentration of LH; sperm production and reserves; and the height of the germinal epithelium of seminiferous tubules. We also observed an early onset of puberty but no effect on the body growth in these animals. These results suggest that maternal exposure to glyphosate disturbed the masculinization process and promoted behavioral changes and histological and endocrine problems in reproductive parameters. These changes associated with the hypersecretion of androgens increased gonadal activity and sperm production.
Assuntos
Glicina/análogos & derivados , Gonadotropinas Hipofisárias/metabolismo , Herbicidas/toxicidade , Exposição Materna/efeitos adversos , Efeitos Tardios da Exposição Pré-Natal/induzido quimicamente , Reprodução/efeitos dos fármacos , Animais , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Estradiol/sangue , Feminino , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Glicina/toxicidade , Hormônio Luteinizante/sangue , Masculino , Preferência de Acasalamento Animal/efeitos dos fármacos , Preferência de Acasalamento Animal/fisiologia , Hipófise/efeitos dos fármacos , Hipófise/metabolismo , Gravidez , Efeitos Tardios da Exposição Pré-Natal/sangue , RNA Mensageiro/metabolismo , Ratos , Reprodução/fisiologia , Epitélio Seminífero/efeitos dos fármacos , Epitélio Seminífero/patologia , Maturidade Sexual/efeitos dos fármacos , Espermatogênese/efeitos dos fármacos , Testosterona/sangue , GlifosatoRESUMO
The present research was performed to isolate and study the effects of a low molecular weight (<1300Da) parasite-associated substance, obtained from peritoneal fluids of female mice infected with Taenia crassiceps cysticerci, on seminiferous epithelium cells of male mice testis. The results showed an intense disruption of Sertoli cells and germ cells within the seminiferous tubules of experimental mice, along with the destruction of their gap junction (GJ). Significant generalized apoptosis of germ cells within seminiferous tubules was determined by TUNEL staining (P=0.0159). In addition, a significant number of infiltrating macrophages were found in the luminal space of these seminiferous tubules (P<0.0001). Finally, electron microscopy studies revealed structural and morphological abnormalities in the somatic cells (Sertoli and Leydig cells) and in the germ cells, primarily in the round and elongate spermatids.
Assuntos
Líquido Ascítico/química , Cisticercose/parasitologia , Cysticercus/metabolismo , Testículo/patologia , Animais , Apoptose , Líquido Ascítico/parasitologia , Cromatografia em Gel , Cisticercose/imunologia , Cisticercose/patologia , Cysticercus/imunologia , Eletroforese em Gel de Poliacrilamida , Feminino , Marcação In Situ das Extremidades Cortadas , Macrófagos/patologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Microscopia Eletrônica de Transmissão , Peso Molecular , Epitélio Seminífero/patologia , Epitélio Seminífero/ultraestrutura , Testículo/ultraestrutura , UltrafiltraçãoRESUMO
BACKGROUND: Amifostine is an efficient cytoprotector against toxicity caused by some chemotherapeutic drugs. Doxorubicin, a potent anticancer anthracycline, is known to produce spermatogenic damage even in low doses. Although some studies have suggested that amifostine does not confer protection to doxorubicin-induced testicular damage, schedules and age of treatment have different approach depending on the protocol. Thus, we proposed to investigate the potential cytoprotective action of amifostine against the damage provoked by doxorubicin to prepubertal rat testes (30-day-old) by assessing some macro and microscopic morphometric parameters 15, 30 and 60 days after the treatment; for fertility evaluation, quantitative analyses of sperm parameters and reproductive competence in the adult phase were also carried out. METHODS: Thirty-day-old male rats were distributed into four groups: Doxorubicin (5 mg/kg), Amifostine (400 mg/kg), Amifostine/Doxorubicin (amifostine 15 minutes before doxorubicin) and Sham Control (0.9% saline solution). "Standard One Way Anova" parametric and "Anova on Ranks" non-parametric tests were applied according to the behavior of the obtained data; significant differences were considered when p < 0.05. RESULTS: The rats killed 30 and 60 days after doxorubicin treatment showed diminution of seminiferous epithelium height and reduction on the frequency of tubular sections containing at least one type of differentiated spermatogonia; reduction of sperm concentration and motility and an increase of sperm anomalous forms where observed in doxorubicin-treated animals. All these parameters were improved in the Amifostine/Doxorubicin group only when compared to Doxorubicin group. Such reduction, however, still remained below the values obtained from the Sham Control group. Nevertheless, the reproductive competence of doxorubicin-treated rats was not improved by amifostine pre-administration. CONCLUSIONS: These results suggest that amifostine promotes a significant reduction of the doxorubicin long-term side effects on the seminiferous epithelium of prepubertal rats, which is reflected in the epidydimal fluid parameters in the adult phase. However, fertility status results suggest that such protection may not be effective against sperm DNA content damage. Further investigation of sperm DNA integrity must be carried out using amifostine and doxorubicin-treated experimental models.
Assuntos
Amifostina/farmacologia , Doxorrubicina/efeitos adversos , Fertilidade/efeitos dos fármacos , Epitélio Seminífero/efeitos dos fármacos , Doenças Testiculares/induzido quimicamente , Doenças Testiculares/prevenção & controle , Animais , Antibióticos Antineoplásicos/efeitos adversos , Citoproteção/efeitos dos fármacos , Regulação para Baixo/efeitos dos fármacos , Feminino , Fertilidade/fisiologia , Nível de Saúde , Masculino , Gravidez , Protetores contra Radiação/farmacologia , Ratos , Ratos Wistar , Análise do Sêmen , Epitélio Seminífero/patologia , Maturidade Sexual/efeitos dos fármacos , Doenças Testiculares/patologiaRESUMO
Organophosphates like O,O-diethyl O-2-isopropyl-6-methyl pyrimidinyl-4-g-1-phosphorothioate (diazinon) are pesticides used worldwide, which can affect both animals and man even after a single exposure. Whereas their toxicity is due to acetylcholinesterase inhibition, their secondary toxic effects have been related to free oxygen radicals. This study evaluates the effects of a single dose of diazinon and melatonin-a powerful antioxidant-on plasmatic acetylcholinesterase activity and testis histopathology in adult mice 1 and 32 days post-treatment. Diazinon diminished the plasma acetylcholinesterase activity on day 1 post-treatment, although testosterone levels remained unaffected. Morphometrical analysis showed a decrease in seminiferous epithelium height (days 1 and 32), whereas an increase in testicular superoxide dismutase (SOD) activity was detected (day 32). Melatonin pretreatment prevented every alteration induced by diazinon, except the diminution of acetylcholinesterase plasmatic activity. Testicular damage might be due to elevated concentrations of free oxygen radicals released upon diazinon exposure, inducing alterations in the DNA and promoting local apoptosis; however, antioxidant pretreatment with melatonin prevents or diminishes this damage.
Assuntos
Antioxidantes/farmacologia , Inibidores da Colinesterase/toxicidade , Diazinon/toxicidade , Inseticidas/toxicidade , Melatonina/farmacologia , Doenças Testiculares/prevenção & controle , Testículo/efeitos dos fármacos , Acetilcolinesterase/sangue , Animais , Colinesterases/sangue , Colinesterases/efeitos dos fármacos , Antagonismo de Drogas , Masculino , Camundongos , Estresse Oxidativo/efeitos dos fármacos , Epitélio Seminífero/efeitos dos fármacos , Epitélio Seminífero/patologia , Superóxido Dismutase/metabolismo , Doenças Testiculares/induzido quimicamente , Doenças Testiculares/metabolismo , Doenças Testiculares/patologia , Testículo/enzimologia , Testículo/patologia , Testosterona/sangueRESUMO
Chronic exposure to ethanol may results in pathophysiologic changes in cellular function. The present work was designed to investigate the morphology of testis submitted to experimental ethanol ingestion. Experimental animals were divided into two groups. The control group (n=23) received a solid diet and tap water and the alcoholic group (n=23) received the same solid diet and ethanol P.A. diluted 20% in water (v/v). After 120 days of treatment, all animals were anesthetized, weighed and sacrificed. Testosterone and luteinizing hormone levels in serum were lower in the alcoholic group than in the control group. Histological and ultrastructural alterations were observed in the testicular alcoholic germinative cells like enormous spaces, lipid droplets accumulation, digestive vacuoles, irregular diameter of the seminiferous tubules and interstitial dilated blood vessels. It was concluded that 20% ethanol provokes lesions on the testis germinative epithelium probably inducing gonadal dysfunction.
Assuntos
Etanol/toxicidade , Infertilidade Masculina/induzido quimicamente , Espermatogônias/efeitos dos fármacos , Espermatogônias/patologia , Testículo/efeitos dos fármacos , Testículo/patologia , Alcoolismo/metabolismo , Alcoolismo/patologia , Alcoolismo/fisiopatologia , Animais , Depressores do Sistema Nervoso Central/toxicidade , Vesículas Citoplasmáticas/efeitos dos fármacos , Vesículas Citoplasmáticas/metabolismo , Vesículas Citoplasmáticas/patologia , Modelos Animais de Doenças , Infertilidade Masculina/patologia , Infertilidade Masculina/fisiopatologia , Metabolismo dos Lipídeos/efeitos dos fármacos , Metabolismo dos Lipídeos/fisiologia , Hormônio Luteinizante/análise , Hormônio Luteinizante/sangue , Masculino , Epitélio Seminífero/efeitos dos fármacos , Epitélio Seminífero/metabolismo , Epitélio Seminífero/patologia , Sigmodontinae/anatomia & histologia , Sigmodontinae/metabolismo , Espermatogônias/metabolismo , Doenças Testiculares/induzido quimicamente , Doenças Testiculares/metabolismo , Doenças Testiculares/patologia , Testículo/metabolismo , Testosterona/análise , Testosterona/sangueRESUMO
Cisplatin is a potent drug used in clinical oncology but causes spermatogenesis damage. Amifostine is a drug used against toxicity caused by ionizing irradiation and chemotherapeutic drugs. Since cisplatin provokes fertility and induces germ cell apoptosis and necrosis, we proposed to evaluate the amifostine cytoprotective action on testes of cisplatin-treated rats. Thirty-day-old prepubertal Wistar rats received a single cisplatin dose of 5 mg/kg and were killed after 3, 6, and 12 hr. The hematoxylin-eosin stained testicular sections were submitted to histological, morphometric, and stereological analysis. The terminal deoxynucleotidyl transferase-mediated deoxyuridinetriphosphate nick end-labeling (TUNEL) method was used to label apoptotic cells. TUNEL-positive and TUNEL-negative germ cells with abnormal nuclear morphology (ANM) were scored. Significant alterations of greater part of the parameters occurred in the cisplatin-treated group (CE) compared to the group that received amifostine before the cisplatin-treatment (ACE); however, testicular weight and volume did not vary between these groups. Tubular diameter was reduced in CE in comparison to ACE rats, while interstitial tissue and lymphatic space volume and volume density were significantly higher in CE rats; interstitial testicular edema probably occurred in cisplatin-treated rats. CE rats showed important histological alterations, which were more accentuated than in ACE rats. The numerical densities of apoptotic germ cells and TUNEL-negative cells with ANM were lower in ACE than in CE rats. In conclusion, the amifostine previously administered to prepubertal rats reduced the testicular damage caused by cisplatin. We conclude that amifostine partially protected the rat seminiferous epithelium against cisplatin toxicity.
Assuntos
Amifostina/farmacologia , Antineoplásicos/antagonistas & inibidores , Antineoplásicos/toxicidade , Cisplatino/antagonistas & inibidores , Cisplatino/toxicidade , Testículo/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Marcação In Situ das Extremidades Cortadas , Masculino , Substâncias Protetoras/farmacologia , Ratos , Ratos Wistar , Epitélio Seminífero/efeitos dos fármacos , Epitélio Seminífero/patologia , Espermatogênese/efeitos dos fármacos , Testículo/patologiaRESUMO
Cyclosporine A (CsA) is known to have testicular toxicity, leading to male infertility. Stimulant and aphrodisiac properties have been attributed to the plant, Heteropterys aphrodisiaca. Thus, the present work was undertaken to evaluate the association of the drug and the medicinal herb in Wistar rats, applying testicular morphometry and ultrastructure. Twenty-four rats were used, divided into four groups: I, control; II, CsA; III, simultaneous use of CsA and H. aphrodisiaca; IV, H. aphrodisiaca. Daily administration by gavage was carried out, during 56 days, of water (sham), CsA in a dose of 15 mg/kg per day and/or H. aphrodisiaca in a dose of 0.5 ml of the infusion prepared with 25 g of roots/100 ml of boiling water. Increased body weight was observed for all groups, but the animals that received only CsA showed the smallest body weight gain. Morphometry showed increased connective tissue volumetric proportion and decreased Leydig cell volumetric proportion in CsA-treated rats. Using transmission electron microscopy, it was possible to ascertain that CsA caused seminiferous epithelium degeneration, resulting in Sertoli cell vacuolization, abnormal round and elongated spermatids and large accumulation of residual cytoplasm at the epithelium border next to the lumen. Expanded intercellular spaces between germ cells were still observed in H. aphrodisiaca-treated rat testes. The administration of H. aphrodisiaca infusion to CsA-treated rats diminished nearly all the CsA-induced damage to the testis ultrastructure, suggesting that H. aphrodisiaca infusion may be used combined with CsA to reduce CsA-induced injuries in the testis.
Assuntos
Ciclosporina/antagonistas & inibidores , Ciclosporina/toxicidade , Malpighiaceae , Fitoterapia , Testículo/efeitos dos fármacos , Animais , Ciclosporina/administração & dosagem , Imunossupressores/administração & dosagem , Imunossupressores/antagonistas & inibidores , Imunossupressores/toxicidade , Células Intersticiais do Testículo/efeitos dos fármacos , Células Intersticiais do Testículo/patologia , Masculino , Microscopia Eletrônica de Transmissão , Extratos Vegetais/administração & dosagem , Ratos , Ratos Wistar , Epitélio Seminífero/efeitos dos fármacos , Epitélio Seminífero/patologia , Epitélio Seminífero/ultraestrutura , Espermatogênese/efeitos dos fármacos , Testículo/patologia , Testículo/ultraestruturaRESUMO
OBJECTIVE: Analyze the changes in the seminiferous epithelium in rats with carbon tetrachloride-induced cirrhosis (CCl4). MATERIALS AND METHODS: Forty-eight male Wistar rats aged 45-50 days, weighing 150-180 grams were used. Twenty-two rats underwent CCl4-induced cirrhosis with CCl4 0.25 mL/Kg weekly intragastrically once a week, during 10 weeks. Additionally, they had a 44% food restriction diet (Group 1). The control group was divided in two subgroups: 13 rats had a 44% food restriction diet and no CCl4 (Group 2) and 10 rats were not submitted to CCl4 or food restriction (Group 3). After 10 weeks, the rats were sacrificed and liver sections were collected for histological analysis. The testicular analysis was carried out to evaluate the frequency of tubules in stages VIII and XIV. RESULTS: The mean rates of stage VIII in animals with food restriction plus CCl4-induced cirrhosis and food restriction without CCl4 were significantly different from animals without either food restriction or CCl4 (18.1 +/- 5.5%, 20.5 +/- 2.5% and 13.4 +/- 3.5%, respectively, p = 0.002). The mean rate of stage VIII in rats with cirrhosis was not significantly different from rats without cirrhosis (18.1 +/- 5.5% and 17.4 +/- 4.6% respectively). The mean frequency of stage XIV in rats with cirrhosis was significantly greater than rats without cirrhosis (4.7 +/- 2.3% and 6.8 +/- 1.9% respectively, p = 0.027). CONCLUSION: Animals with CCl4-induced cirrhosis and food restriction have shown alterations in spermatogenic cycle that were not seen in rats without CCl4-induced cirrhosis and food restriction.
Assuntos
Privação de Alimentos , Cirrose Hepática Experimental/patologia , Epitélio Seminífero/patologia , Animais , Tetracloreto de Carbono , Cirrose Hepática Experimental/induzido quimicamente , Masculino , Ratos , Ratos Wistar , Epitélio Seminífero/efeitos dos fármacosRESUMO
OBJECTIVE: Analyze the changes in the seminiferous epithelium in rats with carbon tetrachloride-induced cirrhosis (CCl4). MATERIALS AND METHODS: Forty-eight male Wistar rats aged 45-50 days, weighing 150-180 grams were used. Twenty-two rats underwent CCl4-induced cirrhosis with CCl4 0.25 mL/Kg weekly intragastrically once a week, during 10 weeks. Additionally, they had a 44 percent food restriction diet (Group 1). The control group was divided in two subgroups: 13 rats had a 44 percent food restriction diet and no CCl4 (Group 2) and 10 rats were not submitted to CCl4 or food restriction (Group 3). After 10 weeks, the rats were sacrificed and liver sections were collected for histological analysis. The testicular analysis was carried out to evaluate the frequency of tubules in stages VIII and XIV. RESULTS: The mean rates of stage VIII in animals with food restriction plus CCl4-induced cirrhosis and food restriction without CCl4 were significantly different from animals without either food restriction or CCl4 (18.1 ± 5.5 percent, 20.5 ± 2.5 percent and 13.4 ± 3.5 percent, respectively, p = 0.002). The mean rate of stage VIII in rats with cirrhosis was not significantly different from rats without cirrhosis (18.1 ± 5.5 percent and 17.4 ± 4.6 percent respectively). The mean frequency of stage XIV in rats with cirrhosis was significantly greater than rats without cirrhosis (4.7 ± 2.3 percent and 6.8 ± 1.9 percent respectively, p = 0.027). CONCLUSION: Animals with CCl4-induced cirrhosis and food restriction have shown alterations in spermatogenic cycle that were not seen in rats without CCl4-induced cirrhosis and food restriction.
Assuntos
Animais , Masculino , Ratos , Cirrose Hepática Experimental/patologia , Epitélio Seminífero/patologia , Privação de Alimentos , Tetracloreto de Carbono , Cirrose Hepática Experimental/induzido quimicamente , Epitélio Seminífero/efeitos dos fármacos , Ratos WistarRESUMO
Cimetidine (Tagamet) is a potent histaminic H2-receptor antagonist, extensively prescribed for ulcers and now available without prescription. Cimetidine is a known testicular toxicant, but its mechanism of action remains uncertain. Rats were treated i.p. with cimetidine either at 50 mg/kg or 250 mg/kg body weight for 59 days. Accessory sex organ weights, but not testis weight, were significantly reduced in the high dose treated groups. FSH levels were significantly elevated in both treated groups, but testosterone levels were unchanged. A high degree of variability characterized testis histology, with most tubules appearing normal and some tubules (15-17%) partially lacking or devoid of germ cells. Morphometry showed that although seminiferous tubule volume was not significantly changed, the volume of peritubular tissue was reduced in the high dose group. There was extensive duplication of the basal lamina, lamina densa in both apparently normal spermatogenic tubules and severely damaged tubules. Apoptotic peritubular myoid cells were also found. TUNEL labeling confirmed extensive apoptotic cell death in peritubular cells, but revealed apoptosis of vascular smooth muscle. Given that 1) peritubular myoid cell apoptosis occurs in apparently normal tubules, that 2) basal lamina disorders are found, and that 3) peritubular cells are lost from the testis, it is suggested that the primary event in cimetidine-related damage is targeted to testicular smooth muscle cells. This is the first in vivo-administered toxicant to be described that targets myoid cells, resulting in abnormal spermatogenesis.
Assuntos
Cimetidina/toxicidade , Antagonistas dos Receptores H2 da Histamina/toxicidade , Epitélio Seminífero/patologia , Doenças Testiculares/induzido quimicamente , Doenças Testiculares/patologia , Animais , Apoptose/efeitos dos fármacos , Hormônios Esteroides Gonadais/sangue , Hormônios Esteroides Gonadais/metabolismo , Marcação In Situ das Extremidades Cortadas , Injeções Intraperitoneais , Masculino , Tamanho do Órgão/efeitos dos fármacos , Tamanho do Órgão/fisiologia , Ratos , Ratos Wistar , Testículo/citologia , Testículo/patologiaRESUMO
The scrotal testes of albino rats aged 35 and 45 days were immersed in water at constant temperatures of 43 degrees C, 44 degrees C, or 45 degrees C for periods of 15-45 min in a special heating device. At an age of 60 days, the rats were mated in individual cages with two primiparous rats each. At an age of 90 days, they were killed and their testes were histologically processed. Rats with testes that had been subjected to heating when the animals were 45 days old showed both alterations of the seminiferous tubules and a decrease in fertilizing capacity. The effect of heat was greater in animals at 45 than at 35 days of age. In heat-treated testes, tubules contained PAS-positive concretion, sometimes engulfed by macrophage-derived giant cells and multinucleate cells derived from spermatids that failed to separate during spermiogenesis. The decrease in testicular volume observed after heat treatment was due mainly to reduced parenchymal volume. Thermic lability of seminiferous stem cells increases with age until adulthood, and recovery from heat injury declines.