RESUMO
Pre- and/or post-natal administrations of di(2-ethylhexyl) phthalate (DEHP) in experimental animals cause alterations in the spermatogenesis. However, the mechanism by which DEHP affects fertility is unknown and could be through alterations in the survival and differentiation of the gonocytes. The aim of the present study was to evaluate the effect of a single administration of DEHP in newborn mice on gonocytic proliferation, differentiation and survival and its long-term effects on seminiferous epithelium and sperm quality. BALB/c mice distributed into Control and DEHP groups were used. Each animal in the DEHP group was given a single dose of 500 mg/Kg at birth. The animals were analyzed at 1, 2, 4, 6, 8, 10 and 70 days postpartum (dpp). Testicular tissues were processed for morphological analysis to determine the different types of gonocytes, differentiation index, seminiferous epithelial alterations, and immunoreactivity to Stra8, Pcna and Vimentin proteins. Long-term evaluation of the seminiferous epithelium and sperm quality were carried out at 70 dpp. The DEHP animal group presented gonocytic degeneration with delayed differentiation, causing a reduction in the population of spermatogonia (Stra8 +) in the cellular proliferation (Pcna+) and disorganization of Vimentin filaments. These events had long-term repercussions on the quality of the seminiferous epithelium and semen. Our study demonstrates that at birth, there is a period that the testes are extremely sensitive to DEHP exposure, which leads to gonocytic degeneration and delay in their differentiation. This situation can have long-term repercussions or permanent effects on the quality of the seminiferous epithelium and sperm parameters.
Assuntos
Animais Recém-Nascidos , Dietilexilftalato , Camundongos Endogâmicos BALB C , Animais , Dietilexilftalato/toxicidade , Masculino , Camundongos , Testículo/efeitos dos fármacos , Testículo/crescimento & desenvolvimento , Espermatogênese/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Plastificantes/toxicidade , Feminino , Epitélio Seminífero/efeitos dos fármacosRESUMO
Cryptorchidism (CO) is a risk factor for the development of testicular germ-cell tumors (TGCT). This is supported by reports showing the persistence of gonocytes in CO patients. These cells are proposed to be related to the development of germ-cell neoplasia in situ (GCNIS), which is considered the precursor stage/lesion of TGCT. Therefore, it is proposed that some patients with CO could express some molecular markers related to TGCT. In this study, we analyzed testicular tissue samples from CO, TGCT, and controls. We determined the expression of POU5F1, PLAP, and KIT by immunohistochemistry and that of the hsa-miR-371-373 cluster, hsa-miR-367, and LATS2, PTEN, and IGFR1 genes by RT-qPCR. We then carried out a bioinformatic analysis to identify other possible candidate genes as tumor biomarkers. We found that 16.7% (2/12) of the CO patients presented increased expression of POU5F1, KIT, PLAP, hsa-miR-371-373, and hsa-miR-367 and decreased expression of LATS2 and IGF1R. Finally, the genes ARID4B, GALNT3, and KPNA6 were identified as other possible candidate tumor biomarkers. This is the first report describing the expression of the hsa-miR-371-373 cluster, hsa-miR-367, LATS2, and IGF1R in the testicular tissues of two CO patients with cells immune-positive to POU5F1, PLAP, and KIT, which is similar to what is observed in TGCT.
RESUMO
Some pediatric patients with cryptorchidism preserve cells with gonocyte characteristics beyond their differentiation period, which could support the theory of the gonocyte as a target for malignancy in the development of testicular neoplasia. One of the key molecules in gonocyte malignancy is represented by microRNAs (miRNAs). The goal of this review is to give an overview of miRNAs, a class of small non-coding RNAs that participate in the regulation of gene expression. We also aim to review the crucial role of several miRNAs that have been further described in the regulation of gonocyte differentiation to spermatogonia, which, when transformed, could give rise to germ cell neoplasia in situ, a precursor lesion to testicular germ cell tumors. Finally, the potential use of miRNAs as diagnostic and prognostic biomarkers in testicular neoplasia is addressed, due to their specificity and sensitivity compared to conventional markers, as well as their applications in therapeutics.
Assuntos
MicroRNAs , Neoplasias Embrionárias de Células Germinativas , Neoplasias Testiculares , Biomarcadores/metabolismo , Criança , Humanos , Masculino , MicroRNAs/genética , MicroRNAs/metabolismo , Neoplasias Embrionárias de Células Germinativas/metabolismo , Espermatogônias/metabolismo , Neoplasias Testiculares/diagnóstico , Neoplasias Testiculares/genética , Neoplasias Testiculares/metabolismoRESUMO
Cryptorchidism (CO) or undescended testicle is an abnormality of male gonadal development that can generate long-term repercussions in men, such as infertility and germ cell neoplasia in situ (GCNIS). The origin of these alterations in humans is not completely clear, due to the absence of an animal model with similar testicular development as in humans with CO. This work intends to describe the testicular histological development of dogs with congenital CO, and determine whether the species could adequately serve as a study model for this pathology in humans. The study was carried out with 36 dogs, equally distributed in two groups: healthy control (CTRL) and CO groups. The contralateral testis to the undescended one in CO group of the animals was considered and analyzed. Each group was subdivided in three stages of development: (1) peripubertal stage (6-8 months), (2) young adult (9-48 months) and (3) senile (49-130 months). Histological development, the presence of cells with gonocyte morphology, cell proliferation, testicular lipoperoxidation and hormonal concentrations of testosterone, estradiol, FSH and LH were evaluated and described. In the cryptorchid testes, the first histological alterations appeared from the first stage of development and were maintained until the senile stage. A pronounced testicular lipoperoxidation occurred only in the second stage of development. The histological alterations due to CO were markedly evident in the young adult stage. Testosterone concentrations witnessed a decrease starting from in the second stage and kept on until the last stage. The contralateral testes of the CO animals showed alterations that positioned them between the control and CO testes. Testicular development of dogs with CO is similar to that of humans. The results of the study suggest that this species could serve as a suitable model for the study of CO in humans.
RESUMO
Studies in laboratory animals have shown that male offspring from dams, exposed to nicotine during pregnancy and postnatal periods, show alterations in fertility, although the origin of this is still uncertain. In this study, we examined in a mouse model if the process of gonocyte maturation to spermatogonia was affected in male offspring from dams with nicotine administration during pregnancy and postnatal periods. BALB/C mice, with and without nicotine administrations in pregnancy and postnatal periods, were studied. The animals were euthanized at 3, 7, 10, 16, and 35 days postpartum (dpp). Testicular tissue samples were processed for histological, ultrastructural, and immunohistochemical studies; and testicular lipoperoxidation was determined. It was observed that in the nicotine-exposed animals, there was increased apoptosis and a reduction in the number of gonocytes that matured to spermatogonia. This gonocyte-spermatogonia maturation reduction was associated with a greater immunoreactivity to nicotinic acetylcholine receptors in the germ cells. Lipoperoxidation was similar in both groups until 16 dpp, with significant reduction at 35 dpp. Our findings suggest that nicotine intake during pregnancy and postnatal periods can affect the process of maturation of gonocytes to spermatogonia and the pool of available spermatogonia for spermatogenesis.
Assuntos
Feto/patologia , Nicotina/toxicidade , Efeitos Tardios da Exposição Pré-Natal/patologia , Espermatogônias/patologia , Animais , Animais Recém-Nascidos , Cotinina/análise , Feminino , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Camundongos Endogâmicos BALB C , Gravidez , Túbulos Seminíferos/efeitos dos fármacos , Túbulos Seminíferos/patologia , Espermatogônias/efeitos dos fármacos , Testículo/patologiaRESUMO
Allelic variants in genes implicated in the development of testicular germ cell tumor (TGCT) could be present in patients with cryptorchidism (CO). Currently; the mechanisms explaining this relationship are still unknown. In this study the common clinical features in patients with CO and TGCT and 6 variants of KIT and AR genes associated to TGCT were analyzed. Population analyzed included 328 individuals: 91 patients with CO; 79 with TGCT, 13 of them with previous CO diagnosis, and 158 healthy males. Of the 13 patients with TGCT and history of CO, one patient (7.7%) presented the heterozygous form of the variant rs121913507 and two patients (15.4%) presented homozygote genotype for the variant rs121913506 in KIT gene. Interestingly, the heterozygous form for the variant rs121913506 of KIT gene was identifying in all of 13 patients. The rs201934623, rs774171864, and rs12014709 variants of the AR gene did not show any clinical association. Our results strongly support that genetic component in CO could be conditioning for the development of TGCT. Notably, KIT gene variants might be determinants in the pathological association between TGCT and CO.
RESUMO
Cryptorchidism (CO) is a risk factor for infertility in men. It is associated with an increase in oxidative stress which alters the differentiation of the gonocytes to spermatogonia. Epigallocatechin-3-gallate (EGCG) is an antioxidant that acts as a free radical scavenger and activates the antioxidant enzymes. The aim of this work was to investigate if EGCG plays a role in the protection of the testicle from alterations generated by CO and its possible mechanism. Male rabbits 7 days old were divided into four groups and distributed as follows: 1) control (C) treated with EGCG vehicle (V) (C/V); 2) C with administration of EGCG from 65 to 120 days postpartum (dpp) (C/EGCG); 3) CO induced by administration of 17ß-estradiol plus EGCG vehicle (CO/V) and 4) CO plus EGCG administration (CO/EGCG). The animals were euthanized at 120 dpp and their testes were processed to evaluate lipid peroxidation, activities of superoxide dismutase (SOD) and catalase (CAT) enzymes as well as serum testosterone (T) concentrations. In addition, the rates of apoptosis, cell proliferation and histological alterations were determined. The CO/EGCG group showed a significant reduction in lipid peroxidation, a significant increase in the anti-oxidant enzyme activities and concentrations of T. Also, there was a significant decrease in the histological alterations, absence of gonocytes and active spermatogenesis when compared with CO/V group. These results show that EGCG reduces lipid peroxidation and increases the activity of the endogenous anti-oxidant system which protects the testes from alterations produced by oxidative stress generated during experimental CO.
Assuntos
Antioxidantes/farmacologia , Catequina/análogos & derivados , Criptorquidismo/tratamento farmacológico , Testículo/efeitos dos fármacos , Animais , Antioxidantes/uso terapêutico , Apoptose/efeitos dos fármacos , Catalase/metabolismo , Catequina/farmacologia , Catequina/uso terapêutico , Estradiol , Imuno-Histoquímica , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Estresse Oxidativo/efeitos dos fármacos , Coelhos , Superóxido Dismutase/metabolismo , Testículo/metabolismo , Testículo/patologiaRESUMO
BACKGROUND: The variation in cystic fibrosis (CF) lung disease not always is explained by the CFTR genotype, so it has become apparent that modifier genes must play a considerable role in the phenotypic heterogeneity of CF, so we investigated the association of allelic variants in modifier genes that modulate the severity of lung function in a group of Mexican patients diagnosed with CF. METHODS: We included 140 CF patients classified according to lung phenotype and analyzed 17 single nucleotide polymorphisms (SNPs) by TaqMan® allelic discrimination. RESULTS: We demonstrated that patients with GG or GC genotype of the allelic variant rs11003125 (MBL2-550) of the MBL2 gene exhibit most of the lung manifestations at an earlier age; and the rs1042713 allelic variant of ADRB2 gene, showed statistical difference only with the age of first spirometry. When we used the dominant model, the MBL2 allele rs11003125 (MBL2-550; p = 0.022, Odds Ratio (OR) 2.87, 95% CI 1.14-7.27) was significantly associated with CF patients as risk factor, and the ADRB2 allele rs1042713 (p.Arg16Gly; p = 0.005, Odds Ratio (OR) 0.37, 95% CI 0.19-0.75) was significantly associated with CF patients as protect factor. CONCLUSIONS: Our findings suggest that the MBL2 and ADRB2 genes exerts an important genetic influence on the lung disease in our patients. Taking into account our results, we insist on not leaving aside this type of studies, since having techniques such as GWAS or WES will be able to advance in achieving a better quality of life for CF patients with severe lung disease.
Assuntos
Fibrose Cística/genética , Fibrose Cística/patologia , Lectina de Ligação a Manose/genética , Receptores Adrenérgicos beta 2/genética , Alelos , Regulador de Condutância Transmembrana em Fibrose Cística/genética , Feminino , Genótipo , Humanos , Pulmão/patologia , Masculino , México , Razão de Chances , Fenótipo , Polimorfismo de Nucleotídeo Único/genética , Qualidade de Vida , Espirometria , Adulto JovemRESUMO
Testicular germ cell cancer (TGCT) is the most common malignancy among young adult males, which has become important due to its increased incidence and mortality in the population worldwide. The etiology is multifactorial. Recent studies have shown some associations between the development of isolated TGCT and certain risk factors, such as exposure to endocrine disruptors, cryptorchidism, and family history of cancer, in order to identify the key pieces in carcinogenesis. Some of the most important findings in recent years is the association of different genes, such as c-KIT/KITLG, expression of the miR-371-373 cluster and protein expression as c-KIT and POU5F1 in the development of this neoplasia, and the identification of new molecular markers as TGFBR3 gene, identifying aberrant methylation patterns in promoter regions of several genes, expression of miR-1297 which regulates PTEN and protein expression as DMTR1. In the future, a multidisciplinary research strategy could provide valuable new insights into the etiology of TGCTs, which support clinical diagnosis of TGCT in the next years to increase survival in this kind of patients.
Assuntos
Neoplasias Embrionárias de Células Germinativas/etiologia , Neoplasias Embrionárias de Células Germinativas/genética , Neoplasias Testiculares/etiologia , Neoplasias Testiculares/genética , Células-Tronco Germinativas Adultas/patologia , Animais , Criptorquidismo/complicações , Meio Ambiente , Epigênese Genética , Predisposição Genética para Doença , Humanos , Masculino , Modelos Biológicos , Mutação , Neoplasias Embrionárias de Células Germinativas/metabolismo , Polimorfismo de Nucleotídeo Único , Proteômica , Fatores de Risco , Neoplasias Testiculares/metabolismoRESUMO
The present study was designed to describe the development of germ cell neoplasia in situ in Chinchilla rabbit by administration of estradiol. The study was performed in rabbits distributed into two groups: control and 17 ß-estradiol. The determination of histological alterations and POU5F1 and c-kit proteins employed as biomarkers for the diagnosis of this neoplasia was carried out. Testicular descent and complete spermatogenesis were observed in the control group. The protein biomarkers were negative. However, in the rabbits treated with estradiol, the testes remained undescended with the gonocytes undifferentiated to spermatogonia. There were histological lesions owing to germ cell neoplasia in situ and positive to POU5F1 and c-kit proteins. These findings indicate that the chinchilla rabbit is an ideal model to study this neoplasia in which the histological characteristics and biomarkers of the disease could be clearly observed. Using this model we suggested that the persisting gonocytes could be responsible for the development of germ cell neoplasia in situ.
Assuntos
Carcinoma in Situ/patologia , Neoplasias Testiculares/patologia , Animais , Biomarcadores Tumorais/análise , Chinchila , Modelos Animais de Doenças , Imuno-Histoquímica , Masculino , Microscopia Eletrônica de Transmissão , Fator 3 de Transcrição de Octâmero/análise , Proteínas Proto-Oncogênicas c-kit/análise , Coelhos , Reação em Cadeia da Polimerase em Tempo RealRESUMO
Ginkgo Biloba extract 761 (EGb 761) is a patented and well-defined mixture of active compounds extracted from Ginkgo biloba leaves. This extract contains two main groups of active compounds, flavonoids (24%) and terpenoids (6%). EGb 761 is used clinically to treat dementia and vaso-occlusive and cochleovestibular disorders. This extract has neuroprotective effects, exerted probably by means of its antioxidant function. Parkinson's disease (PD) is a neurodegenerative disorder that affects 2% of the population older than 60 y. It produces a progressive loss of dopaminergic neurons and depletion of dopamine (DA), leading to movement impairment. The production of reactive oxygen species, which act as mediators of oxidative damage, is linked to PD. This disease is routinely treated with the DA precursor, L-3,4-dihydroxyphenylalanine. However, this produces severe side effects, and its neurotoxic properties can be due to a free radical production. Thus, administration of antioxidant drugs might be used to prevent neuronal death produced by oxidative mechanisms. The use of synthetic antioxidants has decreased because of their suspected activity as carcinogenic promoters. We describe the studies related to the antioxidant effect of EGb 761 in an animal model of PD. It has been shown that EGb761 can provide a neuroprotective/neurorecovery effect against the damage to midbrain DA neurons in an animal model of PD. EGb 761 also has been found to lessen the impairment of locomotion, correlating with an increase of DA and other morphologic and biochemical parameters related to its antioxidant effect in an animal model of PD. These studies suggest it as an alternative in the future treatment of PD.
Assuntos
Antiparkinsonianos/uso terapêutico , Suplementos Nutricionais , Modelos Animais de Doenças , Doença de Parkinson/prevenção & controle , Fitoterapia , Extratos Vegetais/uso terapêutico , Animais , Anti-Inflamatórios não Esteroides/efeitos adversos , Anti-Inflamatórios não Esteroides/química , Anti-Inflamatórios não Esteroides/uso terapêutico , Antioxidantes/efeitos adversos , Antioxidantes/química , Antioxidantes/uso terapêutico , Antiparkinsonianos/efeitos adversos , Antiparkinsonianos/química , Apoptose , Suplementos Nutricionais/efeitos adversos , Suplementos Nutricionais/análise , Neurônios Dopaminérgicos/metabolismo , Ginkgo biloba , Humanos , Mesencéfalo/metabolismo , Fármacos Neuroprotetores/efeitos adversos , Fármacos Neuroprotetores/química , Fármacos Neuroprotetores/uso terapêutico , Estresse Oxidativo , Doença de Parkinson/dietoterapia , Doença de Parkinson/metabolismo , Extratos Vegetais/efeitos adversos , Extratos Vegetais/químicaRESUMO
Cryptorchidism causes apoptosis of germ cells. It has been suggested that the redox regulatory system is involved in this process. The free radicals produced are thought to be generated during the production of uric acid, a reaction catalyzed by xanthine oxidase. This enzyme is inhibited by allopurinol; however, the role of allopurinol in neonate rats with inguinal cryptorchidism has not been assessed yet. Sixty male Wistar rats were used and five groups were formed: a control, a sham, a sham group with allopurinol administration and two groups with surgical unilateral cryptorchidism, which either did not receive, or received, allopurinol. The rats were assessed at 40 days post-partum. Reactive oxygen species concentration and epithelial area were measured and the histopathological, apoptotic and cellular proliferation indexes were determined. We found a decrease in reactive oxygen species, histopathological and apoptotic indexes and an increase in proliferation index and epithelial area in rats with cryptorchidism treated with allopurinol in comparison with rats with untreated cryptorchidism. We suggest that the over-production of reactive oxygen species plays an important role in the damage of the cryptorchid testes. Allopurinol administration decreases reactive oxygen species concentrations as well as the damage to the germ epithelium.
Assuntos
Alopurinol/farmacologia , Criptorquidismo/tratamento farmacológico , Criptorquidismo/patologia , Células Epiteliais/efeitos dos fármacos , Sequestradores de Radicais Livres/farmacologia , Espécies Reativas de Oxigênio/toxicidade , Testículo/efeitos dos fármacos , Alopurinol/administração & dosagem , Análise de Variância , Animais , Animais Recém-Nascidos , Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Criptorquidismo/etiologia , Células Epiteliais/patologia , Masculino , Ratos , Ratos Wistar , Testículo/citologia , Testículo/patologia , Testículo/cirurgiaRESUMO
During the first days of postnatal life in rats the male germ cells (gonocytes) proliferate and move towards the seminiferous tubule basal lamina maturing into spermatogonia. This process is necessary for spermatogenesis and can be affected by estrogen (E); therefore, it is important to determine whether the damaging mechanism induced by E administration during the postnatal period impairs gonocyte maturation. One-day-old rat pups were given 1 microg 17-beta-estradiol daily and studied at 3, 5, 8, 10 and 16 days of age, corresponding to the critical gonocyte differentiation period in the rat. Testicles were isolated and the number of gonocytes in contact with the basal lamina of the seminiferous tubule was estimated, as well as the proliferation rate and apoptosis of the gonocytes. We observed that the administration of E changed the migration of gonocytes towards the basal lamina, decreased cell proliferation and increased apoptosis, resulting in a decrease in spermatogonia and spermatocytes. The migration of gonocytes and subsequent proliferation is required for survival of this germ cell type. The lack of maturation and the death of gonocytes could be one of the causes of infertility following exogenous E treatment.
Assuntos
Estradiol/toxicidade , Espermatozoides/efeitos dos fármacos , Testículo/efeitos dos fármacos , Animais , Animais Recém-Nascidos , Apoptose/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Fertilidade/efeitos dos fármacos , Masculino , Ratos , Ratos Wistar , Espermatozoides/patologia , Testículo/patologiaRESUMO
El intestino delgado de la rata presenta cambios morfológicos y enzimáticos asociados con el destete, los cuales pueden ser alterados por el destete temprano. Sin embargo, el criterio morfométrico no ha sido considerado. Material y Métodos: En este estudio, los efectos del destete precoz (15 días) y prolongado (32 días) sobre el tamaño y número de vellosidades y criptas del intestino delgado, fueron analizados en ratas de 16 a 70 días de edad. Resultados: El destete precoz provocó un incremento temprano en el tamaño de las vellosidades, profundidad y número de criptas en el duodeno y yeyuno, mientras que el número de vellosidades disminuyó. Las crías amamantadas hasta los 32 días no mostraron alteraciones en los parámetros analizados. No obstante, el ileon no presento alteraciones con el destete precoz o prolongado. Conclusiones: Estos datos sostienen el concepto de un programa biológico intrínseco como control de dieta parece tener una función modificadora en el duodeno y el yeyuno