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1.
Chemosphere ; 242: 125285, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31896209

RESUMO

The aim of this study was to assess the long-term effects of synthetic progestin norethindrone (NET) on the growth, reproductive histology, and transcriptional expression profiles of genes associated with the hypothalamic-pituitary-gonadal (HPG) axis and germ cells in adult zebrafish. Adult zebrafish were exposed to 7, 84 and 810 ng/L NET for 90 days. The results showed that exposure to 810 ng/L NET caused a significant decrease in growth of females and males. The ovary weight and GSI was significantly reduced by NET at concentrations of 84 or 810 ng/L, which came along with the delay of ovary maturation in females. However, NET at all treatments resulted in acceleration of sperm maturation in males. In the ovaries of females, a strong inhibition of cyp19a1a gene was observed following exposure to NET at 810 ng/L. Similarly, NET at the highest treatment led to a significant down-regulation of cyp17, cyp19a1a, vasa, nanos1, dazl and dmc1 genes in the testes of males. Taken together, the overall results demonstrated that NET could impact growth and gonadal maturation, with significant alterations of transcriptional expression genes along HPG axis and germ cells.


Assuntos
Expressão Gênica/efeitos dos fármacos , Noretindrona/toxicidade , Progestinas/toxicidade , Poluentes Químicos da Água/toxicidade , Proteínas de Peixe-Zebra/genética , Peixe-Zebra/crescimento & desenvolvimento , Animais , Regulação para Baixo , Feminino , Células Germinativas/efeitos dos fármacos , Gônadas/efeitos dos fármacos , Masculino , Ovário/efeitos dos fármacos , Reprodução/efeitos dos fármacos , Diferenciação Sexual/efeitos dos fármacos , Testículo/efeitos dos fármacos , Peixe-Zebra/genética
2.
Endocr Regul ; 53(2): 93-99, 2019 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-31517623

RESUMO

OBJECTIVE: Stem cell therapy, specifically, pre-induction of mesenchymal stem cells toward male germ-like cells may be useful in patients with azoospermia. The aim of this study was to evaluate in vitro differentiation of mouse bone marrow-derived mesenchymal stem cells (BMSCs) into male germ-like cells by indirect co-culture with testicular cells in the presence of bone morphogenetic protein 4 (BMP4). METHODS: Experimental groups included: control (mouse BMSCs), treatment group-1 (BMSCs treated with BMP4), treatment group-2 (indirect co-culture of BMSCs with mouse testicular cells in the presence of BMP4) and treatment group-3 (indirect co-culture of BMSCs with testicular cells). BMSCs-derived male germ-like cells were evaluated by the expression of Dazl, and Stra8 using RT-qPCR. RESULTS: Stra8 gene expression was significantly increased in the treatment group-2 and Dazl gene was significantly increased in the treatment group-1 compared to other groups. In conclusion, indirect co-culturing of BMSCs with testicular cells and BMP4 leads to the differentiation of BMSCs into male germ-like cells which express specific male germ-like genes. Testicular cells released factors that contributed to the differentiation of BMSCs into male germ progenitor cells. CONCLUSION: This study suggests that mesenchymal stem cells may be differentiated into male germ-like cells and therefore, may be a novel treatment option for men with azoospermia.


Assuntos
Proteína Morfogenética Óssea 4/farmacologia , Diferenciação Celular/efeitos dos fármacos , Células-Tronco Mesenquimais/efeitos dos fármacos , Espermatozoides/efeitos dos fármacos , Testículo/citologia , Animais , Células da Medula Óssea/efeitos dos fármacos , Células da Medula Óssea/fisiologia , Células Cultivadas , Técnicas de Cocultura/métodos , Células Germinativas/efeitos dos fármacos , Células Germinativas/fisiologia , Humanos , Masculino , Células-Tronco Mesenquimais/fisiologia , Camundongos , Espermatozoides/fisiologia
3.
Ecotoxicol Environ Saf ; 183: 109499, 2019 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-31398581

RESUMO

Perfluoroalkyl acids (PFAAs) are anthropogenic compounds used globally in a variety of commercial products. Perfluorononanoic acid (PFNA), a member of PFAAs, is detected in human blood and this has been reported to cause hepatotoxic, immunotoxic, and developmental and testicular toxic effects in laboratory animals. We have recently shown that the acute exposure to PFNA in prepubertal Parkes (P) mice impairs spermatogenesis by inducing oxidative stress and inhibiting testosterone biosynthesis in the testis. The present study was aimed to examine the effect of acute exposure to PFNA in prepubertal P mice on germ cell dynamics and to understand the possible mechanisms of action of this compound on testicular functions. PFNA (2 and 5 mg/kg body weight) was orally administered to male mice for 14 days from postnatal day 25-38. The treatment caused a decrease in overall germ cell transformation. The results also reveal that impairment in testicular functions in treated mice is associated with alterations in cholesterol and glucose homeostasis; further, an inhibition in expressions of growth hormone receptor (GHR), insulin-like growth factor-1 (IGF-1), insulin-like growth factor-1 receptor (IGF-1R), androgen receptor (AR), phosphorylated mammalian target of rapamycin (p-mTOR) and peroxisome proliferator activated receptor α (PPAR α) in the testis is also implicated in this action. The findings thus suggest involvement of multiple factors which altogether contribute to the alterations in spermatogenic process and testosterone production following acute exposure to PFNA in prepubertal mice.


Assuntos
Fluorcarbonetos/toxicidade , Células Germinativas/efeitos dos fármacos , Homeostase/efeitos dos fármacos , Testículo/efeitos dos fármacos , Animais , Glicemia/metabolismo , Colesterol/sangue , Fluorcarbonetos/administração & dosagem , Células Germinativas/metabolismo , Células Germinativas/patologia , Masculino , Camundongos , Testículo/metabolismo , Testes de Toxicidade Aguda
4.
Reprod Toxicol ; 88: 39-47, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31288075

RESUMO

Effects of endocrine disrupting chemicals (EDCs) on reproduction have not been fully explained comprehensively. In this study, we tried to validate the common effect of Bisphenol A (BPA) and Nonylphenol (NP) on the differentiation of embryonic stem (ES) cells and found that they modify the expression of germ cell specific genes. To elucidate functional significance on biological process, we performed Gene Ontology (GO)-based microarray analysis comparing with published GeneChip data of primordial germ cell development in vivo. Cluster analysis of gene expression profile revealed that EDC treatment and primordial germ cell (PGC) development shared characteristic cluster consists of GO terms related to "germ cell development" and "reproduction". In the GO term "reproduction", meiosis related genes showed high expression level by EDC exposure. These results suggest that BPA and NP affect not only some of the germ cell specific genes, but functionally interferes germ cell development and reproduction.


Assuntos
Compostos Benzidrílicos/toxicidade , Células-Tronco Embrionárias/efeitos dos fármacos , Perfilação da Expressão Gênica , Fenóis/toxicidade , Animais , Células Cultivadas , Células Germinativas/efeitos dos fármacos , Células Germinativas/crescimento & desenvolvimento , Camundongos , Camundongos Endogâmicos ICR , Análise de Sequência com Séries de Oligonucleotídeos , Reprodução/efeitos dos fármacos
5.
Sci Total Environ ; 692: 240-248, 2019 Nov 20.
Artigo em Inglês | MEDLINE | ID: mdl-31349165

RESUMO

This study was done on SD rat Sertoli-germ co-cultured cells (Sertoli-germ cells) with nickel nanoparticles (Ni NPs). A series of investigations were performed to observe the role of Ni NPs on the apoptosis of Sertoli-germ cells and to explore the long-chain non-coding RNA (lncRNA) functions on key signaling pathways and regulatory mechanisms. We found that Ni NPs had an apoptotic effect on Sertoli-germ cells. Ni NPs-induced apoptosis in Sertoli-germ cells involves the LOC102551356, Insulin-like growth factor-binding protein 3 (Igfbp3), and mitochondrial apoptosis pathway. The specific mechanism may be: during the process of Ni NPs-induced apoptosis in Sertoli-germ cells, the expression of LOC102551356 is up-regulated, and LOC102551356 activates the mitochondrial apoptosis pathway through targeted regulation of the target gene Igfbp3 in the P53-reduced apoptosis pathway. The results of this study will be important for the safety evaluation of Ni NPs in the future, and could provide an approach for the prevention or alleviation of the toxicity induced by Ni NPs.


Assuntos
Apoptose/efeitos dos fármacos , Células Germinativas/efeitos dos fármacos , Nanopartículas Metálicas/efeitos adversos , Níquel/efeitos adversos , Células de Sertoli/efeitos dos fármacos , Animais , Masculino , RNA Longo não Codificante/metabolismo , Ratos , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacos
6.
Chemosphere ; 235: 70-75, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31255767

RESUMO

Some agrochemicals are genotoxic to several organisms. Nevertheless, few protocols are currently available for measuring the toxicogenetic effects of these compounds in target and non-target field-collected species of insects important to agriculture. Herein, we used the species Ceraeochrysa claveri (Neuroptera: Chrysopidae), a non-target predator insect, to investigate the ability of an azadirachtin-based biopesticide (Azamax™) to induce DNA damage. The alkaline version of the comet assay was standardized to evaluate genetic instability caused by the toxicant in somatic (gut) and germ (nurse cells and oocytes) cells of C. claveri. For this, C. claveri larvae were distributed into three groups (10/each) and treated with Azamax™ at 0, 0.3% or 0.5% throughout the larval stage. DNA damage (tail intensity) was measured in adult insects, four days after emerged. The data showed that both doses of Azamax™ (0.3% and 0.5%) were able to significantly (p < 0.05) increase DNA damage in somatic and germ cells of C. claveri. In conclusion, C. claveri (intestinal and ovarian cells) was a sensitive bioindicator for identifying Azamax™ genotoxic potential, whereas the comet assay was a useful tool for detecting the genotoxic hazard of the pesticide in the field-collected insect species. Given that estimation of adverse effects of pollutants on ecosystems is an essential component of environmental risk assessment, the approach used can be recommended to estimate the ecotoxicity of agricultural chemicals.


Assuntos
Agroquímicos/farmacologia , Insetos/efeitos dos fármacos , Testes de Mutagenicidade/métodos , Praguicidas/toxicidade , Agroquímicos/toxicidade , Animais , Ensaio Cometa/métodos , Dano ao DNA/efeitos dos fármacos , Ecossistema , Células Germinativas/efeitos dos fármacos , Larva/efeitos dos fármacos , Limoninas , Praguicidas/química
7.
Environ Toxicol ; 34(10): 1074-1084, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31157505

RESUMO

Microcystin-LR (MC-LR), a potent endotoxin, can induce reproductive toxicity. In order to investigate the role and mechanisms of apoptosis (p53-dependent and mitochondrial pathways) of germ cells induced by MC-LR, the co-cultured primary Sertoli-germ cells from Sprague-Dawley rats were used for the experiments. Expression levels of proteins, genes, and mitochondrial membrane potential (MMP) were obtained after exposing co-cultured Sertoli-germ cells to MC-LR with or without the addition of the p53 inhibitor, pifithrin-α (PFT-α), and MMP inhibitor, cyclosporin A (CsA). Results indicated that MC-LR could activate p53-dependent pathway-associated proteins in Sertoli-germ cells, leading to a decrease in MMP (indicating the opening of mitochondrial permeability transition pore [mPTP] and the release of Cytochrome-c [Cyt-c]) from the mitochondria into the cytoplasm and eventually the induction of apoptosis. PFT-α inhibited the expression ofp53, ameliorated the MMP of the co-cultured Sertoli-germ cells, and prevented the release of Cyt-c from the mitochondria into the cytoplasm, which reduces the occurrence of apoptosis. Similarly, the decreased release of Cyt-c from the mitochondria into the cytoplasm and the declined level of apoptosis in Sertoli-germ cells induced by MC-LR were observed after the addition of CsA. These results indicated that the apoptosis of the co-cultured Sertoli-germ cells induced by MC-LR was mediated by the p53-dependent pathway, with the involvement of the opening of mPTP.


Assuntos
Células Germinativas/efeitos dos fármacos , Microcistinas/toxicidade , Proteínas de Transporte da Membrana Mitocondrial/metabolismo , Células de Sertoli/efeitos dos fármacos , Proteína Supressora de Tumor p53/metabolismo , Animais , Apoptose/efeitos dos fármacos , Técnicas de Cocultura , Células Germinativas/citologia , Células Germinativas/metabolismo , Masculino , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Ratos , Ratos Sprague-Dawley , Células de Sertoli/citologia , Células de Sertoli/metabolismo
8.
Environ Mol Mutagen ; 60(5): 445-454, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30891817

RESUMO

Drugs taken during pregnancy can affect three generations at once: the gestating woman (F0), her exposed fetus (F1), and the fetal germ cells that confer heritable information for the grandchildren (F2). Unfortunately, despite growing evidence for connections between F0 drug exposures and F2 pathology, current approaches to risk assessment overlook this important dimension of risk. In this commentary, we argue that the unique molecular vulnerabilities of the fetal germline, particularly with regard to global epigenomic reprogramming, combined with empirical evidence for F2 effects of F1 in utero drug and other exposures, should change the way we consider potential long-term consequences of pregnancy drugs and alter toxicology's standard somatic paradigm. Specifically, we (1) suggest that pregnancy drugs common in the postwar decades should be investigated as potential contributors to the "missing heritability" of many pathologies now surging in prevalence; (2) call for inclusion of fetal germline risks in pregnancy drug safety assessment; and (3) highlight the need for intensified research to ascertain generational impacts of diethylstilbestrol, a vanguard question of human germline toxicity. Only by fully addressing this important dimension of transplacental exposure can we responsibly evaluate safety of drug exposures during pregnancy and convey the full scope of risks, while also retrospectively comprehending the generational legacy of recent history's unprecedented glut of evolutionarily novel intrauterine exposures. Environ. Mol. Mutagen. 60:445-454, 2019. © 2019 Wiley Periodicals, Inc.


Assuntos
Células Germinativas/efeitos dos fármacos , Exposição Materna , Troca Materno-Fetal/fisiologia , Efeitos Tardios da Exposição Pré-Natal/patologia , Analgésicos/efeitos adversos , Dietilestilbestrol/efeitos adversos , Estrogênios não Esteroides/efeitos adversos , Feminino , Humanos , Neoplasias/induzido quimicamente , Neoplasias/epidemiologia , Gravidez
9.
Int J Surg Pathol ; 27(5): 568-573, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30907195

RESUMO

Adult neuroblastoma is an extremely infrequent neoplasm, usually occurring in the adrenal medulla or in the paraspinal sympathetic ganglia, as its childhood counterpart. We report a very unusual case of a Schwannian stroma-poor adult neuroblastoma of inguinal location, showing aberrant expression of germ cell markers: SALL4 and OCT4. This aberrant marker expression, the unusual positivity for NKX2.2 and the very scattered (instead of diffuse strong) PHOX2B expression, complicated the initial diagnosis. In this case, the posttreatment histological evaluation revealed the neuroblastic nature of the lesion. Neuroblastoma maturation after treatment is an unusual finding in adults, and in this case, added an important clue for the final diagnosis. Germs cells markers expression in neuroblastoma is an interesting feature to explore and may define a subset of neuroblastomas with a different biological nature.


Assuntos
Neoplasias Abdominais/diagnóstico , Biomarcadores Tumorais/metabolismo , Células Germinativas/patologia , Canal Inguinal/patologia , Neuroblastoma/diagnóstico , Neoplasias Abdominais/patologia , Neoplasias Abdominais/terapia , Adulto , Protocolos de Quimioterapia Combinada Antineoplásica/farmacologia , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Quimiorradioterapia/métodos , Ciclofosfamida/farmacologia , Ciclofosfamida/uso terapêutico , Dactinomicina/farmacologia , Dactinomicina/uso terapêutico , Diagnóstico Diferencial , Etoposídeo/farmacologia , Etoposídeo/uso terapêutico , Células Germinativas/efeitos dos fármacos , Humanos , Ifosfamida/farmacologia , Ifosfamida/uso terapêutico , Canal Inguinal/diagnóstico por imagem , Masculino , Neuroblastoma/patologia , Neuroblastoma/terapia , Fator 3 de Transcrição de Octâmero/metabolismo , Fatores de Transcrição/metabolismo , Vincristina/farmacologia , Vincristina/uso terapêutico
10.
Gen Comp Endocrinol ; 277: 56-65, 2019 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-30878349

RESUMO

Unlike its paralog Foxl2, which is well known for its role in ovarian development in vertebrates, the function of Foxl3 is still unclear. Foxl3 is an ancient duplicated copy of Foxl2. It is present as a single copy in ray-finned fish. But, due to repeated losses, it is absent in most tetrapods. Our transcriptomic data, however, show that two Foxl3s (Foxl3a and its paralog Foxl3b) are present in Japanese eel. Foxl3a is predominantly expressed in the pituitary, and Foxl3b is predominantly expressed in the gills. Both Foxl3s show a sex-dimorphic expression, being higher expression in testes than in ovaries. Moreover, Foxl3a and Foxl3b were exclusively expressed during gonadal differentiation in control eels (100% male). Conversely, Foxl3a and Foxl3b significantly decreased after gonadal differentiation in E2-treated eels (100% female). Furthermore, in accordance the difference in adhesive ability between somatic cells and germline cells in testes, Foxl3s showed a high expression in suspension cells (putative germline cells) and low expression in adhesive cells (putative somatic cells). In situ hybridization further showed that Foxl3a and Foxl3b were expressed in the testicular germline cells. In addition, Foxl3s expression was not changed by sex steroids in in vitro testes culture. Taken together, our results suggest that the teleost-specific Foxl3 paralog was repeatedly lost in most fish after the third round of whole genome duplication. The two germline-expressed Foxl3s had higher expression levels in males than in females during gonadal differentiation in Japanese eel. These results demonstrated that Foxl3s might play an important role in germline sexual fate determination from ancient fish to modern fish.


Assuntos
Anguilla/genética , Anguilla/fisiologia , Fatores de Transcrição Forkhead/genética , Regulação da Expressão Gênica no Desenvolvimento , Células Germinativas/metabolismo , Gônadas/fisiologia , Diferenciação Sexual/fisiologia , Sequência de Aminoácidos , Animais , Tamanho Corporal/efeitos dos fármacos , Estradiol/farmacologia , Fatores de Transcrição Forkhead/química , Fatores de Transcrição Forkhead/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Células Germinativas/efeitos dos fármacos , Gônadas/efeitos dos fármacos , Masculino , Filogenia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Diferenciação Sexual/efeitos dos fármacos , Diferenciação Sexual/genética , Esteroides/farmacologia , Testículo/citologia , Testículo/efeitos dos fármacos , Testículo/metabolismo
11.
Redox Biol ; 22: 101131, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30735911

RESUMO

Chemoresistance is a major therapeutic obstacle in the treatment of human pancreatic ductal adenocarcinoma (PDAC). As an oxidative stress responsive transcription factor, nuclear factor erythroid 2-related factor 2 (Nrf2) regulates the expression of cytoprotective genes. Nrf2 not only plays a critical role in chemoprevention, but also contributes to chemoresistance. In this study, we found that digoxin markedly reversed drug resistance of gemcitabine by inhibiting Nrf2 signaling in SW1990/Gem and Panc-1/Gem cells. Further research revealed that digoxin regulated Nrf2 at transcriptional level. In in vivo study, we found that digoxin and gemcitabine in combination inhibited tumor growth more substantially when compared with gemcitabine treatment alone in SW1990/Gem-shControl cells-derived xenografts. In the meantime, SW1990/Gem-shNrf2 cells-derived xenografts responded to gemcitabine and combination treatment similarly, suggesting that digoxin sensitized gemcitabine-resistant human pancreatic cancer to gemcitabine, which was Nrf2 dependent. These results demonstrated that digoxin might be used as a promising adjuvant sensitizer to reverse chemoresistance of gemcitabine-resistant pancreatic cancer to gemcitabine via inhibiting Nrf2 signaling.


Assuntos
Desoxicitidina/análogos & derivados , Digoxina/farmacologia , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Fator 2 Relacionado a NF-E2/metabolismo , Transdução de Sinais/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Desoxicitidina/farmacologia , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Resistencia a Medicamentos Antineoplásicos/genética , Sinergismo Farmacológico , Perfilação da Expressão Gênica , Células Germinativas/efeitos dos fármacos , Células Germinativas/metabolismo , Humanos , Camundongos , Modelos Biológicos , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transcriptoma , Ensaios Antitumorais Modelo de Xenoenxerto
12.
Toxicol Ind Health ; 35(3): 228-238, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30755103

RESUMO

Di(2-ethylhexyl)phthalate (DEHP) is a typical endocrine-disrupting chemical and reproductive toxicant. Although previous studies have attempted to describe the mechanism by which DEHP exposure results in reproductive dysfunction, few studies focused on puberty, a critical period of reproductive development, and the increased susceptibility to injury in adolescents. To elucidate the mechanism underpinning the testicular effects of DEHP in puberty, we sought to investigate the JAZF1/TR4 pathway in the testes of pubertal rats. Specifically, we focused on the role of the JAZF1/TR4 pathway in male reproduction, including the genes JAZF1, TR4, Sperm 1, and Cyclin A1. In the present study, rats were exposed to increasing concentrations of DEHP (0, 250, 500, and 1000 mg/kg/day) by oral gavages for 30 days. Then we assayed testicular zinc and oxidative stress levels. Our results indicated that DEHP exposure could lead to oxidative stress and decrease the contents of testicular zinc. Additionally, significant morphological changes and cell apoptosis were observed in testes exposed to DEHP, as identified by hematoxylin and eosin staining and the terminal deoxynucleotidyl transferase-mediated nick and labeling assay. By measuring the expression levels of the above relevant genes by qPCR, we found the DEHP-induced increased expression of JAZF1 and decreased expression of TR4, Sperm 1, and Cyclin A1. Therefore, we have demonstrated that in vivo exposure to DEHP might induce reproductive toxicity in pubertal male rats through the JAZF1/TR4 pathway and oxidative stress.


Assuntos
Dietilexilftalato/toxicidade , Estresse Oxidativo/efeitos dos fármacos , Plastificantes/toxicidade , Maturidade Sexual/efeitos dos fármacos , Testículo/efeitos dos fármacos , Animais , Ciclina A1/efeitos dos fármacos , Disruptores Endócrinos/toxicidade , Células Germinativas/efeitos dos fármacos , Masculino , Fatores do Domínio POU/efeitos dos fármacos , Fatores do Domínio POU/genética , Ratos , Ratos Sprague-Dawley , Receptores de Esteroides/efeitos dos fármacos , Receptores de Esteroides/genética , Receptores dos Hormônios Tireóideos/efeitos dos fármacos , Receptores dos Hormônios Tireóideos/genética , Reprodução/efeitos dos fármacos , Testículo/patologia
13.
PLoS Genet ; 15(2): e1007975, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30763314

RESUMO

Chemicals that are highly prevalent in our environment, such as phthalates and pesticides, have been linked to problems associated with reproductive health. However, rapid assessment of their impact on reproductive health and understanding how they cause such deleterious effects, remain challenging due to their fast-growing numbers and the limitations of various current toxicity assessment model systems. Here, we performed a high-throughput screen in C. elegans to identify chemicals inducing aneuploidy as a result of impaired germline function. We screened 46 chemicals that are widely present in our environment, but for which effects in the germline remain poorly understood. These included pesticides, phthalates, and chemicals used in hydraulic fracturing and crude oil processing. Of the 46 chemicals tested, 41% exhibited levels of aneuploidy higher than those detected for bisphenol A (BPA), an endocrine disruptor shown to affect meiosis, at concentrations correlating well with mammalian reproductive endpoints. We further examined three candidates eliciting aneuploidy: dibutyl phthalate (DBP), a likely endocrine disruptor and frequently used plasticizer, and the pesticides 2-(thiocyanomethylthio) benzothiazole (TCMTB) and permethrin. Exposure to these chemicals resulted in increased embryonic lethality, elevated DNA double-strand break (DSB) formation, activation of p53/CEP-1-dependent germ cell apoptosis, chromosomal abnormalities in oocytes at diakinesis, impaired chromosome segregation during early embryogenesis, and germline-specific alterations in gene expression. This study indicates that this high-throughput screening system is highly reliable for the identification of environmental chemicals inducing aneuploidy, and provides new insights into the impact of exposure to three widely used chemicals on meiosis and germline function.


Assuntos
Caenorhabditis elegans/efeitos dos fármacos , Avaliação Pré-Clínica de Medicamentos/métodos , Poluentes Ambientais/toxicidade , Células Germinativas/efeitos dos fármacos , Ensaios de Triagem em Larga Escala/métodos , Aneugênicos/toxicidade , Aneuploidia , Animais , Animais Geneticamente Modificados , Benzotiazóis/toxicidade , Caenorhabditis elegans/embriologia , Caenorhabditis elegans/genética , Quebras de DNA de Cadeia Dupla , Dibutilftalato/toxicidade , Exposição Ambiental , Inseticidas/toxicidade , Meiose/efeitos dos fármacos , Permetrina/toxicidade , Plastificantes/toxicidade , Tiocianatos/toxicidade
14.
Methods Cell Biol ; 150: 429-447, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30777187

RESUMO

Sea urchin gametes have been historically used to demonstrate fertilization and early development in student laboratories. Large amounts of egg and sperm are easily acquired, and the conspicuous changes in egg surface morphology, indicative of sperm-egg fusion and egg activation, are readily observed in the classroom. However, less often incorporated into teaching labs are exercises that demonstrate the dramatic metabolic changes that accompany egg activation. One example is the massive up-regulation of various essential transport activities in the embryo's plasma membrane, including xenobiotic transporter activity. Here we outline a laboratory that incorporates this concept into a teaching lab, capitalizing on the magnitude and uniformity of the xenobiotic transporter activation event in certain species of sea urchins. The introduction of this chapter provides background information for the instructor, and the remainder serves as a laboratory manual for students. The experiments detailed within the manual can be completed in a total of 4-8h spread over one or two lab periods. The lab manual guides students through a modified version of the United States Environmental Protection Agency (EPA) toxicity test, a novel undergraduate-level laboratory on xenobiotic transporters, and analysis of microscope data using ImageJ. We have found this lab to be of interest to a wide range of biology and environmental science undergraduates, and effective in teaching underlying concepts in developmental biology, physiology and toxicology.


Assuntos
Fertilização/efeitos dos fármacos , Ouriços-do-Mar/efeitos dos fármacos , Xenobióticos/administração & dosagem , Animais , Transporte Biológico/efeitos dos fármacos , Membrana Celular/metabolismo , Biologia do Desenvolvimento/métodos , Embrião de Mamíferos/efeitos dos fármacos , Células Germinativas/efeitos dos fármacos
15.
Biomed Pharmacother ; 111: 568-578, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30597310

RESUMO

BACKGROUND: Diabetes is one of the most chronic and widespread diseases causing the damages to the male reproductive system. Nowadays, several studies have been performed to show the role of phenolic compounds in reducing the complications of diabetes. Carvacrol is a phenolic monoterpene which has been shown to have much therapeutic efficacy in various diseases. METHODS: Thirty-two adult male Wistar rats (n = 8 in each group) were used in this experimental study. The induction of diabetes was performed using a single intraperitoneal (IP) injection of STZ (50 mg/kg). Rats were assigned into the following groups: control group, diabetic group, diabetic group daily fed with carvacrol at a dose of 75 mg/kg for 8 weeks, and the control group daily fed with carvacrol at a dose of 75 mg/kg for 8 weeks. RESULTS: Treatment with carvacrol significantly improved the histological morphology of the testis, reduced the tissue activity of superoxide dismutase (SOD) and glutathione peroxidase (GPx) enzymes, and diminished the elevated levels of tissue malondialdehyde (MDA) (p < 0.05). Moreover, our results showed that carvacrol significantly decreased Bax and increased Bcl-2 at the levels of gene and protein expression. It also significantly (p < 0.05) reduced the rate of germ cell apoptosis. CONCLUSION: It seems that the treatment with carvacrol mitigates testicular tissue damage in diabetic rats possibly through its antioxidant properties.


Assuntos
Antioxidantes/uso terapêutico , Diabetes Mellitus Experimental/tratamento farmacológico , Células Germinativas/efeitos dos fármacos , Monoterpenos/uso terapêutico , Estresse Oxidativo/efeitos dos fármacos , Testículo/efeitos dos fármacos , Fatores Etários , Animais , Antioxidantes/farmacologia , Apoptose/efeitos dos fármacos , Apoptose/fisiologia , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Experimental/patologia , Células Germinativas/metabolismo , Células Germinativas/patologia , Masculino , Monoterpenos/farmacologia , Estresse Oxidativo/fisiologia , Ratos , Ratos Wistar , Testículo/metabolismo , Testículo/patologia , Resultado do Tratamento
16.
Toxicol Lett ; 303: 48-54, 2019 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-30599193

RESUMO

Our goal was to study the effect of BP3 (benzophenone 3) in the follicular assembly and the potential involvement of Foxl2 pathway using whole ovary cultures. Ovaries were collected from Wistar rats at birth, treated in vitro with vehicle (0.01% DMSO), BP3 (5.8 nM, 276 nM, 576 nM and 876 nM) or ESR2 inhibitor (0.1 nM), and cultured for 7 days. Nest breakdown, follicular assembly and the expression of several regulators of these processes (p27, Foxl2, Sox9, Bmp2, Cyp19 and Fst) were evaluated. In vitro exposure to BP3 (5.8 nM) decreased the population of total oocytes, the number of nests per ovary and early primary follicles population. In addition, BP3 (5.8 nM) induced overexpression of Foxl2 mRNA levels through ESR2 but increased Fst mRNA levels independently from ESR2 or Foxl2. We also observed that the number of p27-positive oocytes was decreased after BP3 (5.8 nM). On the other hand, exposure to BP3-276 increased total oocytes, the number of nests per ovary and decreased primary follicles. In addition, BP3-276 induced no changes of Foxl2 mRNA levels through ESR2 but increased Fst mRNA levels independently from ESR2 or Foxl2. In conclusion, our study clearly shows that exposure to BP3 is to perturb the early events of germ cell development as showed here in whole ovary cultures.


Assuntos
Benzofenonas/toxicidade , Folículo Ovariano/efeitos dos fármacos , Animais , Proteína Morfogenética Óssea 2/genética , Proteína Morfogenética Óssea 2/metabolismo , Diferenciação Celular/efeitos dos fármacos , Inibidor de Quinase Dependente de Ciclina p27/genética , Inibidor de Quinase Dependente de Ciclina p27/metabolismo , Família 19 do Citocromo P450/genética , Família 19 do Citocromo P450/metabolismo , Feminino , Proteína Forkhead Box L2/genética , Proteína Forkhead Box L2/metabolismo , Regulação da Expressão Gênica , Células Germinativas/efeitos dos fármacos , Células Germinativas/crescimento & desenvolvimento , Oócitos/efeitos dos fármacos , Oócitos/metabolismo , Folículo Ovariano/citologia , Folículo Ovariano/metabolismo , Ratos , Ratos Wistar , Fatores de Transcrição SOX9/genética , Fatores de Transcrição SOX9/metabolismo , Técnicas de Cultura de Tecidos
17.
EMBO J ; 38(1)2019 01 03.
Artigo em Inglês | MEDLINE | ID: mdl-30257965

RESUMO

An intricate link is becoming apparent between metabolism and cellular identities. Here, we explore the basis for such a link in an in vitro model for early mouse embryonic development: from naïve pluripotency to the specification of primordial germ cells (PGCs). Using single-cell RNA-seq with statistical modelling and modulation of energy metabolism, we demonstrate a functional role for oxidative mitochondrial metabolism in naïve pluripotency. We link mitochondrial tricarboxylic acid cycle activity to IDH2-mediated production of alpha-ketoglutarate and through it, the activity of key epigenetic regulators. Accordingly, this metabolite has a role in the maintenance of naïve pluripotency as well as in PGC differentiation, likely through preserving a particular histone methylation status underlying the transient state of developmental competence for the PGC fate. We reveal a link between energy metabolism and epigenetic control of cell state transitions during a developmental trajectory towards germ cell specification, and establish a paradigm for stabilizing fleeting cellular states through metabolic modulation.


Assuntos
Diferenciação Celular/efeitos dos fármacos , Células-Tronco Embrionárias/efeitos dos fármacos , Células Germinativas/efeitos dos fármacos , Ácidos Cetoglutáricos/farmacologia , Células-Tronco Pluripotentes/efeitos dos fármacos , Animais , Diferenciação Celular/genética , Células Cultivadas , Embrião de Mamíferos , Células-Tronco Embrionárias/fisiologia , Epigênese Genética/efeitos dos fármacos , Epigênese Genética/genética , Feminino , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Células Germinativas/fisiologia , Ácidos Cetoglutáricos/metabolismo , Masculino , Redes e Vias Metabólicas/efeitos dos fármacos , Redes e Vias Metabólicas/genética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Células-Tronco Pluripotentes/fisiologia
18.
Pediatr Surg Int ; 35(1): 137-143, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30386894

RESUMO

BACKGROUND: Exposure to ionizing radiation results in cytotoxic and genotoxic effects caused mainly by the oxidative damage. In the present study, we investigated the radioprotective effect of novel antioxidant cocktail on germ cell apoptosis and spermatogenesis in rats subjected to whole body radiation (WBIR). METHODS: Adult male rats weighing 250-270 g were divided into four groups, eight rats each. Group 1 served as untreated control, group 2 received an IP single dose of antioxidant cocktail (1 ml). Group 3 was exposed to a WBIR (6 Gy). Group 4 received antioxidant cocktail before WBIR. Rats from each group were killed after 48 h. MDA levels were measured in serum (TBARS assay). Johnsen's criteria and the number of germinal cell layers were used to categorize spermatogenesis. TUNEL assay was used to determine germ cell apoptosis. Statistical analysis was performed using one-way ANOVA test. RESULTS: WBIR resulted in histological testicular damage (decrease in Johnsen's criteria, p < 0.05) that was accompanied by a significant increase in germ cell apoptosis, expressed as the number of apoptotic cells per 100 tubules (AI-1 apoptotic index) and the number of positive tubules per 100 tubules (AI-2 apoptotic index). Treatment with antioxidant cocktail resulted in a significant decrease in germ cell apoptosis (33% decrease in AI-1, p < 0.05 and 34% decrease in AI-2, p < 0.05) that was accompanied by an improved spermatogenesis (increase in Johnsen's criteria, p < 0.05). CONCLUSIONS: In a rat model of WBIR, antioxidant treatment ameliorates oxidative stress-induced testicular damage, decreases germ cell apoptosis and improves spermatogenesis.


Assuntos
Antioxidantes/farmacologia , Apoptose/efeitos dos fármacos , Células Germinativas/efeitos dos fármacos , Espermatogênese/efeitos dos fármacos , Animais , Células Germinativas/patologia , Células Germinativas/efeitos da radiação , Masculino , Lesões Experimentais por Radiação , Radiação Ionizante , Ratos , Ratos Sprague-Dawley , Espermatogênese/efeitos da radiação , Testículo/efeitos dos fármacos , Testículo/patologia , Testículo/efeitos da radiação
19.
Gene ; 688: 140-150, 2019 Mar 10.
Artigo em Inglês | MEDLINE | ID: mdl-30529510

RESUMO

Moringa oleifera (Moringaceae) is a plant known for having high antioxidant potency, anticancer, hepatoprotective, cardioprotective etc. and many more activities. Besides these, Moringaceae has the potential for attenuating the male sexual dysfunction. Reactive oxygen species/ROS were increased in cryptorchidism and therefore cause infertility by damaging sperm DNA and germ cell apoptosis. There was an increase in heat shock proteins (HSP) in cells, which is affected by heat shock. In the present study, the antioxidant effects of two different doses of M. oleifera Lam Extract (MOLE) on experimentally induced cryptorchid testes of rats was investigated. Forty two male rats (16 days old) were divided into four groups: a normal control group, a cryptorchidism-induced control group and two cryptorchidism-induced groups treated orally with either 400 or 800 mg/kg MOLE for 2 weeks. Our study showed that there were ruptures from interstitial spaces, separation of the germ cells from basal membrane, falling of the germ cells into the lumen, perivascular fibrosis, oedema, increased level of HSP70, apoptosis, malondialdehyde (MDA) and decrease in the level of superoxide dismutase (SOD) after the cryptorchidism. We found that pathological damages, oxidative stress, expression of the HSP70 and germ cell apoptosis were decreased in treated groups with MOLE. In brief, we can say that aqueous extract of M. oleifera reduces the oxidative stress in a unilateral cryptorchidism induced rats, and it might attenuate histopathological damages, HSP expression and germ cell apoptosis.


Assuntos
Apoptose/efeitos dos fármacos , Células Germinativas/efeitos dos fármacos , Proteínas de Choque Térmico HSP70/metabolismo , Moringa oleifera/química , Extratos Vegetais/farmacologia , Animais , Antioxidantes/farmacologia , Criptorquidismo/metabolismo , Células Germinativas/metabolismo , Masculino , Malondialdeído/metabolismo , Modelos Animais , Estresse Oxidativo/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Espermatozoides/efeitos dos fármacos , Espermatozoides/metabolismo , Superóxido Dismutase/metabolismo
20.
Drug Chem Toxicol ; 42(5): 502-508, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29482370

RESUMO

In the present study, coffee (CF) was evaluated for its protective effects against genotoxic damage and oxidative stress induced by the chemotherapeutic drug, cyclophosphamide (CPH). The sex-linked recessive lethal (SLRL) test was employed to study the induction of mutations in the larvae as well as in all the successive germ cell stages of treated males. Control and treated third instar larvae were used to monitor the biomarkers of oxidative stress response such as glutathione content (GSH), glutathione-S-transferase (GST), catalase (CAT), superoxide dismutase (SOD) and lipid peroxidation (MDA content). Our results demonstrated that co-administration of CF (2%) with CPH (3 mM) has significantly reduced CPH-induced lethal mutations in the germ cells of larvae and adult flies. The reductions observed in mutation frequencies were: 75% in larvae and 62.4% in the adult. Significant enhancement in antioxidant enzymatic levels: CAT (46.6%) > SOD (43.0%) > GST (42.4%) > GSH (31.6%) and reduction in MDA levels (32.05%) in the pretreated third instar larvae demonstrated the antioxidant activity of CF against CPH-induced oxidative stress. The findings from the present study suggest that the Drosophila model is an ideal one for evaluating the antigenotoxic and antioxidant activity of complex mixtures like CF.


Assuntos
Antimutagênicos/farmacologia , Café/química , Ciclofosfamida/toxicidade , Dano ao DNA/efeitos dos fármacos , Drosophila melanogaster , Células Germinativas/efeitos dos fármacos , Animais , Antioxidantes/metabolismo , Drosophila melanogaster/genética , Células Germinativas/patologia , Masculino , Testes de Mutagenicidade , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/genética
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