RESUMO
The implementation of Zinc oxide nanoparticles (ZnO NPs) raises concerns regarding their potential toxic effects on human health. Although more and more researches have confirmed the toxic effects of ZnO NPs, limited attention has been given to their impact on the early embryonic nervous system. This study aimed to explore the impact of exposure to ZnO NPs on early neurogenesis and explore its underlying mechanisms. We conducted experiments here to confirm the hypothesis that exposure to ZnO NPs causes neural tube defects in early embryonic development. We first used mouse and chicken embryos to confirm that ZnO NPs and the Zn2+ they release are able to penetrate the placental barrier, influence fetal growth and result in incomplete neural tube closure. Using SH-SY5Y cells, we determined that ZnO NPs-induced incomplete neural tube closure was caused by activation of various cell death modes, including ferroptosis, apoptosis and autophagy. Moreover, dissolved Zn2+ played a role in triggering widespread cell death. ZnO NPs were accumulated within mitochondria after entering cells, damaging mitochondrial function and resulting in the over production of reactive oxygen species, ultimately inducing cellular oxidative stress. The N-acetylcysteine (NAC) exhibits significant efficacy in mitigating cellular oxidative stress, thereby alleviating the cytotoxicity and neurotoxicity brought about by ZnO NPs. These findings indicated that the exposure of ZnO NPs in early embryonic development can induce cell death through oxidative stress, resulting in a reduced number of cells involved in early neural tube closure and ultimately resulting in incomplete neural tube closure during embryo development. The findings of this study could raise public awareness regarding the potential risks associated with the exposure and use of ZnO NPs in early pregnancy.
Assuntos
Desenvolvimento Embrionário , Defeitos do Tubo Neural , Tubo Neural , Estresse Oxidativo , Espécies Reativas de Oxigênio , Óxido de Zinco , Óxido de Zinco/toxicidade , Animais , Estresse Oxidativo/efeitos dos fármacos , Embrião de Galinha , Desenvolvimento Embrionário/efeitos dos fármacos , Camundongos , Tubo Neural/efeitos dos fármacos , Tubo Neural/embriologia , Tubo Neural/metabolismo , Humanos , Defeitos do Tubo Neural/induzido quimicamente , Defeitos do Tubo Neural/metabolismo , Defeitos do Tubo Neural/embriologia , Defeitos do Tubo Neural/patologia , Espécies Reativas de Oxigênio/metabolismo , Apoptose/efeitos dos fármacos , Morte Celular/efeitos dos fármacos , Feminino , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Nanopartículas Metálicas/toxicidade , Autofagia/efeitos dos fármacos , Linhagem Celular Tumoral , Nanopartículas/toxicidadeRESUMO
Neural tube defects (NTDs), which are caused by impaired embryonic neural tube closure, are one of the most serious and common birth defects. Peptidyl-prolyl cis/trans isomerase 1 (Pin1) is a prolyl isomerase that uniquely regulates cell signaling by manipulating protein conformation following phosphorylation, although its involvement in neuronal development remains unknown. In this study, we explored the involvement of Pin1 in NTDs and its potential mechanisms both in vitro and in vivo. The levels of Pin1 expression were reduced in NTD models induced by all-trans retinoic acid (Atra). Pin1 plays a significant role in regulating the apoptosis, proliferation, differentiation, and migration of neurons. Moreover, Pin1 knockdown significantly was found to exacerbate oxidative stress (OS) and endoplasmic reticulum stress (ERs) in neuronal cells. Further studies showed that the Notch1-Nrf2 signaling pathway may participate in Pin1 regulation of NTDs, as evidenced by the inhibition and overexpression of the Notch1-Nrf2 pathway. In addition, immunofluorescence (IF), co-immunoprecipitation (Co-IP), and GST pull-down experiments also showed that Pin1 interacts directly with Notch1 and Nrf2. Thus, our study suggested that the knocking down of Pin1 promotes NTD progression by inhibiting the activation of the Notch1-Nrf2 signaling pathway, and it is possible that this effect is achieved by disrupting the interaction of Pin1 with Notch1 and Nrf2, affecting their proteostasis. Our research identified that the regulation of Pin1 by retinoic acid (RA) and its involvement in the development of NTDs through the Notch1-Nrf2 axis could enhance our comprehension of the mechanism behind RA-induced brain abnormalities.
Assuntos
Peptidilprolil Isomerase de Interação com NIMA , Defeitos do Tubo Neural , Tretinoína , Animais , Feminino , Humanos , Camundongos , Apoptose/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Regulação para Baixo/efeitos dos fármacos , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Tubo Neural/metabolismo , Tubo Neural/efeitos dos fármacos , Defeitos do Tubo Neural/metabolismo , Defeitos do Tubo Neural/genética , Defeitos do Tubo Neural/induzido quimicamente , Neurônios/metabolismo , Neurônios/efeitos dos fármacos , Fator 2 Relacionado a NF-E2/metabolismo , Fator 2 Relacionado a NF-E2/genética , Peptidilprolil Isomerase de Interação com NIMA/metabolismo , Peptidilprolil Isomerase de Interação com NIMA/genética , Estresse Oxidativo/efeitos dos fármacos , Receptor Notch1/metabolismo , Receptor Notch1/genética , Transdução de Sinais/efeitos dos fármacos , Tretinoína/metabolismo , Tretinoína/farmacologiaRESUMO
AIM: To investigate the effects of pregabalin on neural tube closure, and other potential effects on other organ systems in a chick embryo model. MATERIAL AND METHODS: Fertilized chicken eggs were divided into groups, and different doses of pregabalin was administered. All embryos were harvested in the 8th day of incubation, and investigated both macroscopically and microscopically against any developmental malformations caused by Pregabalin. RESULTS: Macroscopically not any malformations were detected but macrosomia was statistically significant in medium and high dose groups. Microscopically, vertebral lamina ossification was delayed in some embryos in high dose group but not interpreted as midline closure defect and also not statistically significant. Decrease in the number of renal glomerulus and increase in the tubular damage was statistically significant in medium and high dose groups. Cardiomegaly was also found in some embryos in middle and high dose groups but not statistically significant. CONCLUSION: The use of pregabalin does not cause neural tube closure defect in the embryo unless not exceed recommended maximum dose. Causing macrosomia instead of developmental retardation by Pregabalin is in conflict with the literature. This study revealed that Pregabalin causes fetal nephrotoxicity and macrosomia. These findings indicate that the use of Pregabalin in pregnancy still needs to be accounted as suspicious.
Assuntos
Desenvolvimento Embrionário/efeitos dos fármacos , Tubo Neural/efeitos dos fármacos , Pregabalina/toxicidade , Teratogênese/efeitos dos fármacos , Animais , Embrião de Galinha , Galinhas/crescimento & desenvolvimento , Relação Dose-Resposta a Droga , Tubo Neural/embriologia , Tubo Neural/crescimento & desenvolvimento , Defeitos do Tubo Neural/induzido quimicamente , Pregabalina/farmacologia , Testes de ToxicidadeRESUMO
AIM: To investigate the effects of quetiapine exposure on neural tube development in early stage chicken embryos. MATERIAL AND METHODS: Eighty-four fertilised specific pathogen-free chicken eggs were divided into four equal groups (groups 1?4). Three experimental groups (groups 2, 3 and 4) and a single control group (group 1) were used. Each egg in group 2 (n=21) was injected with 20 ?L of saline after 30 hours of incubation. Eggs in groups 3 and 4 were injected with 0.02 ml of a solution containing 400 and 800 ?g of quetiapine dose, respectively. Incubation was continued until the end of 72 hours. All embryos were then removed from the eggs and histopathologically examined. RESULTS: Normal development and the closed neural tubes were shown in 18, 16, 13 and 9 embryos in groups 1 2, 3 and 9, respectively, of the 84 embryos incubated. Open neural tubes were found in one, three and five embryos in groups 2, 3 and 5, respectively. Also, developmental anomalies were found in three, four, five and seven embryos in groups 1, 2, 3 and 4, respectively. Moreover, no significant relationship between NTD and quetiapine exposure had been found. CONCLUSION: Quetiapine has no significant effect on the occurrence of neural tube defects in the chicken embryo model.
Assuntos
Antipsicóticos/administração & dosagem , Desenvolvimento Embrionário/efeitos dos fármacos , Tubo Neural/efeitos dos fármacos , Tubo Neural/embriologia , Fumarato de Quetiapina/administração & dosagem , Animais , Antipsicóticos/efeitos adversos , Embrião de Galinha , Galinhas , Desenvolvimento Embrionário/fisiologia , Defeitos do Tubo Neural/induzido quimicamente , Defeitos do Tubo Neural/diagnóstico , Fumarato de Quetiapina/efeitos adversosRESUMO
Nuclear factor kappa B (NF-κB) is a heterodimer of protein subunits p65 and p50, that regulates the expression of a large number of genes related to cell growth and proliferation. The p65 subunit is activated after phosphorylation by Pim-1, while the p50 subunit is the cleaved product of its precursor molecule p105. Valproic acid (VPA), an antiepileptic drug, is a known teratogen and its exposure during pregnancy is associated with 1-2% of neural tube defects in the offspring. The current study aimed at investigating the effects of in utero VPA exposure on the key components of the NF-κB signaling pathway including p65, p50, and Pim-1 in CD-1 mouse embryos during the critical period of neural tube closure. Here we report that p65, Pim-1 and p105/p50 mRNA were significantly (p < 0.05) downregulated at 1 and 3 h following in utero exposure to a teratogenic dose (400 mg/kg) of VPA in gestational day (GD)9 exposed embryos. At GD13 heads of control, non-exencephalic and exencephalic embryos were used for analysis and we found significant upregulation of p65 protein expression in non-exencephalic GD13 heads while p50 protein levels were significantly downregulated in both non-exencephalic and exencephalic groups. On the other hand, p65 and p50 protein levels remained unchanged in the nuclear extracts of the VPA-exposed non-exencephalic and exencephalic GD13 embryo heads. The reported results suggest that VPA exposure perturbates p65, p105/p50, Pim-1 transcript and p65/p50 protein levels in mouse embryos.
Assuntos
NF-kappa B/metabolismo , Tubo Neural/efeitos dos fármacos , Tubo Neural/embriologia , Ácido Valproico/toxicidade , Animais , Anticonvulsivantes/administração & dosagem , Anticonvulsivantes/toxicidade , Modelos Animais de Doenças , Feminino , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Masculino , Troca Materno-Fetal , Camundongos , NF-kappa B/genética , Subunidade p50 de NF-kappa B/genética , Subunidade p50 de NF-kappa B/metabolismo , Tubo Neural/metabolismo , Defeitos do Tubo Neural/induzido quimicamente , Defeitos do Tubo Neural/embriologia , Defeitos do Tubo Neural/metabolismo , Neurotoxinas/administração & dosagem , Neurotoxinas/toxicidade , Neurulação/efeitos dos fármacos , Neurulação/fisiologia , Gravidez , Proteínas Proto-Oncogênicas c-pim-1/genética , Proteínas Proto-Oncogênicas c-pim-1/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Transdução de Sinais/efeitos dos fármacos , Fator de Transcrição RelA/genética , Fator de Transcrição RelA/metabolismo , Ácido Valproico/administração & dosagemRESUMO
BACKGROUND: Early gestational alcohol exposure is associated with severe craniofacial and CNS dysmorphologies and behavioral abnormalities during adolescence and adulthood. Alcohol exposure during the formation of the neural tube (gestational day [GD] 8 to 10 in mice; equivalent to4th week of human pregnancy) disrupts development of ventral midline brain structures such as the pituitary, septum, and ventricles. This study identifies transcriptomic changes in the rostroventral neural tube (RVNT), the region of the neural tube that gives rise to the midline structures sensitive to alcohol exposure during neurulation. METHODS: Female C57BL/6J mice were administered 2 doses of alcohol (2.9 g/kg) or vehicle 4 hours apart on GD 9.0. The RVNTs of embryos were collected 6 or 24 hours after the first dose and processed for RNA-seq. RESULTS: Six hours following GD 9.0 alcohol exposure (GD 9.25), over 2,300 genes in the RVNT were determined to be differentially regulated by alcohol. Enrichment analysis determined that PAE affected pathways related to cell proliferation, p53 signaling, ribosome biogenesis, and immune activation. In addition, over 100 genes involved in primary cilia formation and function and regulation of morphogenic pathways were altered 6 hours after alcohol exposure. The changes to gene expression were largely transient, as only 91 genes identified as differentially regulated by prenatal alcohol at GD 10 (24 hours postexposure). Functionally, the differentially regulated genes at GD 10 were related to organogenesis and cell migration. CONCLUSIONS: These data give a comprehensive view of the changing landscape of the embryonic transcriptome networks in regions of the neural tube that give rise to brain structures impacted by a neurulation-stage alcohol exposure. Identification of gene networks dysregulated by alcohol will help elucidate the pathogenic mechanisms of alcohol's actions.
Assuntos
Depressores do Sistema Nervoso Central/farmacologia , Embrião de Mamíferos/efeitos dos fármacos , Etanol/farmacologia , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Tubo Neural/efeitos dos fármacos , Neurulação/efeitos dos fármacos , Animais , Proliferação de Células/genética , Cílios/genética , Embrião de Mamíferos/metabolismo , Feminino , Perfilação da Expressão Gênica , Camundongos , Tubo Neural/metabolismo , Neurulação/genética , Biogênese de Organelas , Gravidez , RNA-Seq , Ribossomos/genética , Proteína Supressora de Tumor p53RESUMO
AIM: To investigate the effects of tartrazine exposure on neural tube development, in early stage chicken embryos. MATERIAL AND METHODS: A total of 120 fertilized specific pathogen-free chicken eggs were divided into 4 equal groups (groups 1?4). After 30 hours of incubation, the eggs, except for the Group 1 (control group), were opened under 4X optical magnification. Group 2 was administered physiological saline. Group 3 was administered a middle dose of tartrazin (4.5 mg/kg) at a volume of 20 µL by the in ovo method, and group 4 was administered a high dose of tartrazine (7.5 mg/kg) using the same process. Incubation was continued until the end of the 72nd hour; all embryos were then removed from the eggs and histopathologically examined. RESULTS: Of the 120 embryos incubated, normal development and the closed neural tubes were shown in all embryos in group 1; 23 in group 2; 19 in group 3 and; only 9 in group 4. Open neural tubes were found in; 4 embryos in group 2; 5 embryos in group 3 and; 13 embryos in group 4. The neural tube closure defect was found to be significantly higher in group 4 compared to the other groups (p < 0.01). CONCLUSION: Based on our data, tartrazine, as one of the widely used food coloring agent, was seen to cause a neural tube defect in the chicken embryo model.
Assuntos
Corantes de Alimentos/toxicidade , Defeitos do Tubo Neural/induzido quimicamente , Defeitos do Tubo Neural/patologia , Tubo Neural/efeitos dos fármacos , Tartrazina/toxicidade , Animais , Embrião de Galinha , Galinhas , Desenvolvimento Embrionário/efeitos dos fármacos , Desenvolvimento Embrionário/fisiologia , Tubo Neural/patologiaRESUMO
OBJECTIVE: Erythrosine (E127), a synthetic food dye containing iodine and sodium, has often been used inside packaged foods and beverages in Turkey and many other countries. We evaluated the effects of erythrosine on neural tube development in early-stage chicken embryos. METHODS: The study included 4 groups, with a total of 80 embryos: a control group, a normal saline group, a half-dose group, and a high-dose group. After 30 hours of incubation, saline and erythrosine solution was injected under the embryonic discs. At the end of 72 hours, the embryos were excised and evaluated macroscopically and histopathologically. RESULTS: Neural tube defects were detected in the erythrosine-administered groups with statistically significant differences. In contrast, the embryos in the control and saline groups displayed normal development. CONCLUSIONS: Erythrosine increased the risk of neural tube defects in early-stage chicken embryos, even at half of the approved dose.
Assuntos
Eritrosina/farmacologia , Corantes Fluorescentes/farmacologia , Defeitos do Tubo Neural/embriologia , Tubo Neural/efeitos dos fármacos , Anormalidades Induzidas por Medicamentos/embriologia , Anormalidades Induzidas por Medicamentos/etiologia , Animais , Embrião de Galinha , Desenvolvimento Embrionário/efeitos dos fármacos , Tubo Neural/embriologia , Defeitos do Tubo Neural/induzido quimicamenteRESUMO
AIM: To investigate the effects of Phenyramidol (Phe) on neural development in an early chicken embryo model. MATERIAL AND METHODS: Sixty fertile non-pathogenic Super Nick eggs were incubated for 24 hours (h) and divided into four groups of 15 eggs each. Phe was administrated through the sub-blastoderm, and the eggs were incubated for another 24 h. All eggs were opened after 48 h of incubation, and the embryos were evaluated morphologically and histopathologically. RESULTS: In Group 1 (control group), none exhibited neural tube defects (NTDs) (0%), 1 (6.6%) was undeveloped; in Group 2 (low dosages), 1 did not develop (6.6%); in Group 3 (normal dosages), 2 (13.4%) had NTDs, 1 (6.6%) was undeveloped; in Group 4 (high dosages), 5 (33.3%) had NTDs, 2 (13.3%) were undeveloped. CONCLUSION: In light of the results, it was determined that the use of increasing doses of Phe led to defects in midline closure in early chicken embryos. This is the first report in the literature on Phe used in an early chicken embryo model.
Assuntos
Desenvolvimento Embrionário/efeitos dos fármacos , Relaxantes Musculares Centrais/toxicidade , Tubo Neural/efeitos dos fármacos , Tubo Neural/embriologia , Piridinas/toxicidade , Animais , Embrião de Galinha , Galinhas , Desenvolvimento Embrionário/fisiologia , Defeitos do Tubo Neural/induzido quimicamente , Defeitos do Tubo Neural/patologiaRESUMO
BACKGROUND: Neural tube defects are congenital malformations of the central nervous system. Genetic predisposition and some environmental factors play an important role in the development of neural tube defects. This study aimed to investigate the effects of diclofenac sodium on the neural tube development in a chick embryo model that corresponds to the first month of vertebral deve- lopment in mammals. MATERIALS AND METHODS: Seventy-five fertile, specific pathogen-free eggs were incubated for 28 h and were divided into five groups of 15 eggs each. Diclofenac sodium was administered via the sub-blastodermic route at this stage. Incubation was continued till the end of the 48th h. All eggs were then opened and embryos were dissected from embryonic membranes and evaluated morphologically and histopathologically. RESULTS: It was determined that the use of increasing doses of diclofenac sodium led to defects of midline closure in early chicken embryos. There were statistically significant differences in neural tube positions (open or close) among the groups. In addition; crown-rump length, somite number were significantly decreased in high dose experimental groups compared with control group. CONCLUSIONS: This study showed that development of neurons is affected in chi- cken embryos after administration of diclofenac sodium. The exact teratogenic mechanism of diclofenac sodium is not clear; therefore it should be investigated.
Assuntos
Diclofenaco/efeitos adversos , Tubo Neural/embriologia , Animais , Embrião de Galinha , Desenvolvimento Embrionário/efeitos dos fármacos , Tubo Neural/efeitos dos fármacos , Tubo Neural/patologia , Defeitos do Tubo Neural/embriologia , Defeitos do Tubo Neural/patologia , Estatística como AssuntoRESUMO
AIM: To investigate the impact of rizatriptan on neural tube development using early chick embryos as a model organism. MATERIAL AND METHODS: A total of 36 pathogen-free Leghorn chicken eggs were selected and categorized in three groups: sham, therapeutic, and supra-therapeutic. After 24 hours, the eggs were opened and injected with sterile drugs, and then reclosed using plastic tape. After a period of 72 hours, the eggs were opened and assessed using the Hamburger-Hamilton chick embryology classification method. TUNEL staining was used to identify apoptosis, and hematoxylin-eosin staining was used to investigate neural tube closure. RESULTS: Treatment with rizatriptan significantly slowed down neural tube development. The supra-therapeutic group showed neural tube closure defects. CONCLUSION: Rizatriptan had a negative effect on neural tube closure. Further research is needed to identify a safe and effective drug for treating migraines during pregnancy.
Assuntos
Desenvolvimento Embrionário/efeitos dos fármacos , Defeitos do Tubo Neural/induzido quimicamente , Tubo Neural/efeitos dos fármacos , Agonistas do Receptor de Serotonina/toxicidade , Triazóis/toxicidade , Triptaminas/toxicidade , Animais , Embrião de Galinha , Galinhas , Tubo Neural/embriologiaAssuntos
Suplementos Nutricionais , Formiatos/uso terapêutico , Defeitos do Tubo Neural/genética , Tubo Neural/metabolismo , Aminoidrolases/genética , Animais , Modelos Animais de Doenças , Ácido Fólico/genética , Ácido Fólico/metabolismo , Formiato-Tetra-Hidrofolato Ligase/genética , Formiatos/metabolismo , Humanos , Proteínas de Membrana Transportadoras/genética , Metilenotetra-Hidrofolato Desidrogenase (NADP)/genética , Metilenotetra-Hidrofolato Redutase (NADPH2)/genética , Camundongos , Complexos Multienzimáticos/genética , Tubo Neural/efeitos dos fármacos , Tubo Neural/crescimento & desenvolvimento , Tubo Neural/patologia , Defeitos do Tubo Neural/dietoterapia , Defeitos do Tubo Neural/metabolismo , Defeitos do Tubo Neural/patologiaRESUMO
Thymidylate synthase (TYMS) is a key enzyme in the de novo synthesis of 2'-deoxythymidine-5'-monophosphate (dTMP) from 2'-deoxyuridine-5'-monophosphate (dUMP). Our aim was to investigate the role of dTMP dysmetabolism via inhibition of TYMS by an inhibitor, 5-fluorouracil (5-FU) in the occurrence of neural tube defects (NTDs). We found that a high incidence of NTDs occurred after treatment with 5-FU at 12.5â¯mg/kg body weight. TYMS activity was significantly inhibited with decreased dTMP and accumulation of dUMP after 5-FU injection. The proliferation of neuroepithelial cells were markedly inhibited in NTDs compared with control. Expressions of proliferating cell nuclear antigen and phosphohistone H3 were significantly decreased in NTDs, while phosphorylated replication protein A2, P53 and Caspase3 were significantly increased in NTDs compared with control. These results indicated that inhibition of TYMS affected the balance between proliferation and apoptosis in neuroepithelial cells, which might shed some lights on the mechanisms involved in NTDs.
Assuntos
Desenvolvimento Embrionário/efeitos dos fármacos , Defeitos do Tubo Neural/enzimologia , Tubo Neural/efeitos dos fármacos , Timidilato Sintase/antagonistas & inibidores , Animais , Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Nucleotídeos de Desoxiuracil/metabolismo , Fluoruracila/toxicidade , Camundongos Endogâmicos C57BL , Tubo Neural/embriologia , Defeitos do Tubo Neural/induzido quimicamente , Defeitos do Tubo Neural/embriologia , Defeitos do Tubo Neural/patologia , Células Neuroepiteliais/efeitos dos fármacos , Células Neuroepiteliais/enzimologia , Células Neuroepiteliais/patologia , Timidina/análogos & derivados , Timidina/metabolismoRESUMO
Folate functions as a coenzyme to transfer one-carbon units that are necessary for deoxythymidylate synthesis, purine synthesis, and various methylation reactions. Ingested folate becomes a functional molecule through intestinal absorption, circulation, transport to cells, and various modifications to its structure. Associations between nutritional folate status and chronic diseases such as cardiovascular disease, cancer, and cognitive dysfunction have been reported. It has also been reported that maternal folate nutritional status is related to the risk of neural tube defects (NTDs) in the offspring. It has also been recommended that folate be consumed in the diet to promote the maintenance of good health. To reduce the risk of NTDs, supplementation with folic acid (a synthetic form of folate) during the periconceptional period has also been recommended. This paper describes the basic features and nutritional role of folate.
Assuntos
Suplementos Nutricionais , Deficiência de Ácido Fólico/prevenção & controle , Ácido Fólico/metabolismo , Defeitos do Tubo Neural/prevenção & controle , Tubo Neural/metabolismo , Embrião de Mamíferos , Feminino , Feto , Ácido Fólico/administração & dosagem , Deficiência de Ácido Fólico/diagnóstico , Deficiência de Ácido Fólico/metabolismo , Deficiência de Ácido Fólico/patologia , Humanos , Redes e Vias Metabólicas/fisiologia , Tubo Neural/anormalidades , Tubo Neural/efeitos dos fármacos , Tubo Neural/embriologia , Defeitos do Tubo Neural/diagnóstico , Defeitos do Tubo Neural/metabolismo , Defeitos do Tubo Neural/patologia , Recomendações NutricionaisRESUMO
The recapitulation of tissue development and patterning in three-dimensional (3D) culture is an important dimension of stem cell research. Here, we describe a 3D culture protocol in which single mouse ES cells embedded in Matrigel under neural induction conditions clonally form a lumen containing, oval-shaped epithelial structure within 3 days. By Day 7 an apicobasally polarized neuroepithelium with uniformly dorsal cell identity forms. Treatment with retinoic acid at Day 2 results in posteriorization and self-organization of dorsal-ventral neural tube patterning. Neural tube organoid growth is also supported by pure laminin gels as well as poly(ethylene glycol) (PEG)-based artificial extracellular matrix hydrogels, which can be fine-tuned for key microenvironment characteristics. The rapid generation of a simple, patterned tissue in well-defined culture conditions makes the neural tube organoid a tractable model for studying neural stem cell self-organization.
Assuntos
Células-Tronco Embrionárias/citologia , Tubo Neural/citologia , Organogênese/fisiologia , Animais , Técnicas de Cultura de Células/métodos , Células Cultivadas , Colágeno/administração & dosagem , Combinação de Medicamentos , Desenvolvimento Embrionário/efeitos dos fármacos , Desenvolvimento Embrionário/fisiologia , Células-Tronco Embrionárias/efeitos dos fármacos , Matriz Extracelular/fisiologia , Hidrogéis/administração & dosagem , Laminina/administração & dosagem , Camundongos , Tubo Neural/efeitos dos fármacos , Organogênese/efeitos dos fármacos , Organoides/citologia , Polietilenoglicóis/administração & dosagem , Proteoglicanas/administração & dosagem , Tretinoína/farmacologiaRESUMO
Susceptibility to neural tube defects (NTDs), such as anencephaly and spina bifida is influenced by genetic and environmental factors including maternal nutrition. Maternal periconceptional supplementation with folic acid significantly reduces the risk of an NTD-affected pregnancy, but does not prevent all NTDs, and "folic acid non-responsive" NTDs continue to occur. Similarly, among mouse models of NTDs, some are responsive to folic acid but others are not. Among nutritional factors, inositol deficiency causes cranial NTDs in mice while supplemental inositol prevents spinal and cranial NTDs in the curly tail (Grhl3 hypomorph) mouse, rodent models of hyperglycemia or induced diabetes, and in a folate-deficiency induced NTD model. NTDs also occur in mice lacking expression of certain inositol kinases. Inositol-containing phospholipids (phosphoinositides) and soluble inositol phosphates mediate a range of functions, including intracellular signaling, interaction with cytoskeletal proteins, and regulation of membrane identity in trafficking and cell division. Myo-inositol has been trialed in humans for a range of conditions and appears safe for use in human pregnancy. In pilot studies in Italy and the United Kingdom, women took inositol together with folic acid preconceptionally, after one or more previous NTD-affected pregnancies. In nonrandomized cohorts and a randomized double-blind study in the United Kingdom, no recurrent NTDs were observed among 52 pregnancies reported to date. Larger-scale fully powered trials are needed to determine whether supplementation with inositol and folic acid would more effectively prevent NTDs than folic acid alone. Birth Defects Research 109:68-80, 2017. © 2016 The Authors Birth Defects Research Published by Wiley Periodicals, Inc.
Assuntos
Suplementos Nutricionais , Deficiência de Ácido Fólico/prevenção & controle , Ácido Fólico/administração & dosagem , Inositol/administração & dosagem , Defeitos do Tubo Neural/prevenção & controle , Tubo Neural/efeitos dos fármacos , Animais , Ensaios Clínicos como Assunto , Modelos Animais de Doenças , Feminino , Deficiência de Ácido Fólico/genética , Deficiência de Ácido Fólico/metabolismo , Deficiência de Ácido Fólico/patologia , Humanos , Fosfatos de Inositol/metabolismo , Fenômenos Fisiológicos da Nutrição Materna , Camundongos , Tubo Neural/anormalidades , Tubo Neural/metabolismo , Defeitos do Tubo Neural/genética , Defeitos do Tubo Neural/metabolismo , Defeitos do Tubo Neural/patologia , Fosfatidilinositóis/metabolismo , GravidezRESUMO
BACKGROUND: Neural tube defects are debilitating birth defects that occur when the developing neural plate fails to close in early gestation. Arsenic induces neural tube defects in animal models, but whether environmental arsenic exposure increases risk of neural tube defects in humans is unknown. METHODS: We describe a new case-control study in Bangladesh, a country currently experiencing an epidemic of arsenic poisoning through contaminated drinking water. We plan to understand how arsenic influences risk of neural tube defects in humans through mechanisms that include disruption of maternal glucose and folate metabolism, as well as epigenetic effects. We also investigate whether sweat chloride concentration, a potential new biomarker for arsenic toxicity, can be used to identify women at higher risk for having a child affected by neural tube defect. We will collect dural tissue from cases, obtained at the time of surgical closure of the defect, and believe investigation of these samples will provide insight into the epigenetic mechanisms by which prenatal arsenic exposure affects the developing nervous system. CONCLUSION: These studies explore mechanisms by which arsenic may increase risk of neural tube defects in humans and use a unique population with high arsenic exposure to test hypotheses. If successful, these studies may assist countries with high arsenic exposure such as Bangladesh to identify populations at high risk of neural tube defects, as well as direct development of novel screening strategies for maternal risk.Birth Defects Research 109:92-98, 2017.© 2016 The Authors Birth Defects Research Published by Wiley Periodicals, Inc.
Assuntos
Intoxicação por Arsênico/diagnóstico , Arsênio/toxicidade , Cloretos/análise , Ácido Fólico/metabolismo , Glucose/metabolismo , Meningomielocele/diagnóstico , Poluentes Químicos da Água/toxicidade , Intoxicação por Arsênico/etiologia , Intoxicação por Arsênico/metabolismo , Intoxicação por Arsênico/patologia , Bangladesh , Biomarcadores/análise , Estudos de Casos e Controles , Criança , Exposição Ambiental/efeitos adversos , Feminino , Humanos , Meningomielocele/etiologia , Meningomielocele/metabolismo , Meningomielocele/patologia , Tubo Neural/anormalidades , Tubo Neural/efeitos dos fármacos , Tubo Neural/metabolismo , Risco , Suor/químicaRESUMO
BACKGROUND: Periconception maternal nutrition and folate in particular are important factors influencing the incidence of neural tube defects (NTDs). Many but not all NTDs are prevented by folic acid supplementation and there is a pressing need for additional strategies to prevent these birth defects. Other micronutrients such as iron are potential candidates, yet a clear role for iron deficiency in contributing to NTDs is lacking. Our previous studies with the flatiron (ffe) mouse model of Ferroportin1 (Fpn1) deficiency suggest that iron is required for neural tube closure and forebrain development raising the possibility that iron supplementation could prevent NTDs. METHODS: We determined the effect of periconception iron and/or folic acid supplementation on the penetrance of NTDs in the Fpn1ffe mouse model. Concurrently, measurements of folate and iron were made to ensure supplementation had the intended effects. RESULTS: High levels of iron supplementation significantly reduced the incidence of NTDs in Fpn1ffe mutants. Fpn1 deficiency resulted in reduced folate levels in both pregnant dams and embryos. Yet folic acid supplementation did not prevent NTDs in the Fpn1ffe model. Similarly, forebrain truncations were rescued with iron. Surprisingly, the high levels of iron supplementation used in this study caused folate deficiency in wild-type dams and embryos. CONCLUSION: Our results demonstrate that iron supplementation can prevent NTDs and forebrain truncations in the Fpn1ffe model. Surprisingly, high levels of iron supplementation and iron overload can cause folate deficiency. If iron is essential for neural tube closure, it is possible that iron deficiency might contribute to NTDs. Birth Defects Research 109:81-91, 2017. © 2016 The Authors Birth Defects Research Published by Wiley Periodicals, Inc.
Assuntos
Proteínas de Transporte de Cátions/genética , Suplementos Nutricionais , Deficiência de Ácido Fólico/dietoterapia , Ácido Fólico/administração & dosagem , Ferro/administração & dosagem , Defeitos do Tubo Neural/prevenção & controle , Animais , Proteínas de Transporte de Cátions/deficiência , Cruzamentos Genéticos , Modelos Animais de Doenças , Embrião de Mamíferos , Feminino , Deficiência de Ácido Fólico/genética , Deficiência de Ácido Fólico/metabolismo , Deficiência de Ácido Fólico/patologia , Deleção de Genes , Humanos , Fenômenos Fisiológicos da Nutrição Materna , Camundongos , Camundongos Transgênicos , Tubo Neural/anormalidades , Tubo Neural/efeitos dos fármacos , Tubo Neural/metabolismo , Defeitos do Tubo Neural/genética , Defeitos do Tubo Neural/metabolismo , Defeitos do Tubo Neural/patologia , Penetrância , Gravidez , Prosencéfalo/anormalidades , Prosencéfalo/efeitos dos fármacos , Prosencéfalo/metabolismoRESUMO
BACKGROUND: Valproic acid (VPA) is prescribed therapeutically for multiple conditions, including epilepsy. When taken during pregnancy, VPA is teratogenic, increasing the risk of several birth and developmental defects including neural tube defects (NTDs). The mechanism by which VPA causes NTDs remains controversial and how VPA interacts with folic acid (FA), a vitamin commonly recommended for the prevention of NTDs, remains uncertain. We sought to address both questions by applying untargeted metabolite profiling analysis to neural tube closure (NTC) stage mouse embryos. METHODS: Pregnant SWV dams on either a 2 ppm or 10 ppm FA supplemented diet were injected with a single dose of VPA on gestational day E8.5. On day E9.5, the mouse embryos were collected and evaluated for NTC status. Liquid chromatography coupled to mass spectrometry metabolomics analysis was performed to compare metabolite profiles of NTD-affected VPA-exposed whole mouse embryos with profiles from embryos that underwent normal NTC from control dams. RESULTS: NTDs were observed in all embryos from VPA-treated dams and penetrance was not diminished by dietary FA supplementation. The most profound metabolic perturbations were found in the 10ppm FA VPA-exposed mouse embryos, compared with the other three treatment groups. Affected metabolites included amino acids, nucleobases and related phosphorylated nucleotides, lipids, and carnitines. CONCLUSION: Maternal VPA treatment markedly perturbed purine and pyrimidine metabolism in E9.5 embryos. In combination with a high FA diet, VPA treatment resulted in gross metabolic changes, likely caused by a multiplicity of mechanisms, including an apparent disruption of mitochondrial beta-oxidation. Birth Defects Research 109:106-119, 2017. © 2016 Wiley Periodicals, Inc.
Assuntos
Suplementos Nutricionais , Ácido Fólico/administração & dosagem , Defeitos do Tubo Neural/metabolismo , Neurulação/efeitos dos fármacos , Teratogênicos/toxicidade , Ácido Valproico/toxicidade , Aminoácidos/metabolismo , Animais , Carnitina/metabolismo , Modelos Animais de Doenças , Embrião de Mamíferos , Feminino , Lipídeos/análise , Masculino , Metaboloma , Camundongos , Tubo Neural/anormalidades , Tubo Neural/efeitos dos fármacos , Tubo Neural/metabolismo , Defeitos do Tubo Neural/induzido quimicamente , Defeitos do Tubo Neural/patologia , Penetrância , Gravidez , Purinas/metabolismo , Pirimidinas/metabolismoRESUMO
Sonic hedgehog (SHH) es un morfógeno esencial para el desarrollo del tubo neural, miembros y somitos. Variaciones en su expresión pueden ocasionar alteraciones en el sistema nervioso. Esto lo producen teratógenos, como el ácido valproico (VPA), el cual aumenta las especies reactivas de oxígeno, pudiendo contrarrestarse con la administración de vitamina E (VE). Se buscó determinar la expresión de SHH en tubo neural y médula espinal en embriones y fetos de ratones expuestos a VPA, VPA + VE y VE. Se conformaron 8 grupos de ratones hembra (Mus musculus). A los 8 días post-coito (p.c.) se les administró a los grupos 1 y 5 suero fisiológico 0,3 ml; grupos 2 y 6 VPA 600 mg/kg; grupos 3 y 7 VPA 600 mg/kg + VE 200 UI/kg; grupos 4 y 8 VE 200 UI/kg, todos los tratamientos vía oral. A los 12 días p.c., se sacrificaron los grupos 1, 2, 3 y 4, y a los 17 días los restantes. Fueron fijados en solución Bouin e incluidos en paraplast. Se realizaron cortes transversales a nivel torácico. Se utilizó anticuerpo policlonal anti-SHH (Santa Cruz, H-160, conejo), dilución 1:100. Se describió la morfología de las muestras marcadas positivamente, se midió la densidad óptica integrada y porcentaje de área inmunoreactiva. La expresión de SHH fue inmunopositiva en notocorda y placa del piso del tubo neural solo en embriones de 12 días p.c. Los grupos tratados con VPA+VE y VE presentaron mayor intensidad inmunohistoquímica y porcentaje de área inmunoreactiva en comparación al grupo tratado con VPA (p 0,0001) en la placa del piso, siendo similar al grupo control. En la notocorda, la intensidad de inmunoreacción fue similar a lo demostrado en la placa del piso, con diferencias significativas (p 0,0001), pero el porcentaje de área no arrojó diferencias. Los grupos de 17 días de gestación resultaron negativos a la expresión de SHH. La vitamina E regula la expresión de SHH en tubo neural, atenuando así los efectos del VPA.
Sonic hedgehog (SHH) is an essential morphogen for the development of neural tube, members and somites. Variations in expression can cause abnormalities in the nervous system. This will produce teratogens, such as valproic acid (VPA), which increases the reactive oxygen species and can be counteracted with the administration of vitamin E (VE). We sought to determine the expression of SHH in the neural tube and spinal cord in mice embryos and fetuses exposed to VPA, VPA + VE and VE. For the study we used 8 groups of female mice (Mus musculus). At day 8 post-coitus (p.c.) the groups were administered as follows: groups 1 and 5,0.3ml saline; groups 2 and 6, VPA 600 mg/kg; groups 3 and 7, VPA 600 mg / kg + VE 200 IU/kg; groups 4 and 8, VE 200 IU/kg, all treatments were given orally. On the 12th day p.c., groups 1, 2, 3 and 4 were euthanized and the remaining groups at day 17. They were fixed in Bouin solution and included in paraplast; thoracic cross sections were performed, anti-SHH polyclonal antibody (Santa Cruz, H-160, rabbit) dilution 1:100 was used. We described morphology of the positively labeled samples and measured integrated optic density and percentage of immunoreactivearea.SHH expression was immunopositive in notochord and floor plate of the neural tube in embryos only 12 day p.c. In the groups treated with VPA + VE and VE immunohistochemistry showed greater intensity and percentage of immunoreactive area compared to those in the group treated with VPA (p0.0001) in the floor plate, being similar to the control group. In the notochord, immunoreaction intensity was similar to that shown in the floor plate, with significant differences (p 0.0001), but the percentage of area showed no differences. The groups at day 17 of gestation were negative for the expression of SHH. VE regulates expression of SHH in neural tube, thus attenuating the effects of VPA.