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1.
Sci Data ; 11(1): 586, 2024 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-38839806

RESUMO

Sex bias is known in the prevalence/pathology of neurodevelopmental disorders. Sex-dependent differences of the certain brain areas are known to emerge perinatally through the exposure to sex hormones, while gene expression patterns in the rodent embryonic brain does not seem to be completely the same between male and female. To investigate potential sex differences in gene expression and cortical organization during the embryonic period in mice, we conducted a comprehensive analysis of gene expression for the telencephalon at embryonic day (E) 11.5 (a peak of neural stem cell expansion) and E14.5 (a peak of neurogenesis) using bulk RNA-seq data. As a result, our data showed the existence of notable sex differences in gene expression patterns not obviously at E11.5, but clearly at E14.5 when neurogenesis has become its peak. These data can be useful for exploring potential contribution of genes exhibiting sex differences to the divergence in brain development. Additionally, our data underscore the significance of studying the embryonic period to gain a deeper understanding of sex differences in brain development.


Assuntos
Telencéfalo , Transcriptoma , Animais , Telencéfalo/embriologia , Telencéfalo/metabolismo , Camundongos , Feminino , Masculino , Neurogênese/genética , Caracteres Sexuais
2.
Biomolecules ; 13(5)2023 04 27.
Artigo em Inglês | MEDLINE | ID: mdl-37238632

RESUMO

22q11.2 deletion syndrome (22q11.2DS) is associated with a high risk of developing various psychiatric and developmental disorders, including schizophrenia and early-onset Parkinson's disease. Recently, a mouse model of this disease, Del(3.0Mb)/+, mimicking the 3.0 Mb deletion which is most frequently found in patients with 22q11.2DS, was generated. The behavior of this mouse model was extensively studied and several abnormalities related to the symptoms of 22q11.2DS were found. However, the histological features of their brains have been little addressed. Here we describe the cytoarchitectures of the brains of Del(3.0Mb)/+ mice. First, we investigated the overall histology of the embryonic and adult cerebral cortices, but they were indistinguishable from the wild type. However, the morphologies of individual neurons were slightly but significantly changed from the wild type counterparts in a region-specific manner. The dendritic branches and/or dendritic spine densities of neurons in the medial prefrontal cortex, nucleus accumbens, and primary somatosensory cortex were reduced. We also observed reduced axon innervation of dopaminergic neurons into the prefrontal cortex. Given these affected neurons function together as the dopamine system to control animal behaviors, the impairment we observed may explain a part of the abnormal behaviors of Del(3.0Mb)/+ mice and the psychiatric symptoms of 22q11.2DS.


Assuntos
Síndrome de DiGeorge , Doença de Parkinson , Esquizofrenia , Animais , Camundongos , Síndrome de DiGeorge/genética , Síndrome de DiGeorge/complicações , Síndrome de DiGeorge/diagnóstico , Esquizofrenia/patologia , Encéfalo/patologia , Doença de Parkinson/patologia , Córtex Pré-Frontal
3.
EMBO Rep ; 22(2): e51524, 2021 02 03.
Artigo em Inglês | MEDLINE | ID: mdl-33399271

RESUMO

Advanced paternal age can have deleterious effects on various traits in the next generation. Here, we establish a paternal-aging model in mice to understand the molecular mechanisms of transgenerational epigenetics. Whole-genome target DNA methylome analyses of sperm from aged mice reveal more hypo-methylated genomic regions enriched in REST/NRSF binding motifs. Gene set enrichment analyses also reveal the upregulation of REST/NRSF target genes in the forebrain of embryos from aged fathers. Offspring derived from young mice administrated with a DNA de-methylation drug phenocopy the abnormal vocal communication of pups derived from aged fathers. In conclusion, hypo-methylation of sperm DNA can be a key molecular feature modulating neurodevelopmental programs in offspring by causing fluctuations in the expression of REST/NRSF target genes.


Assuntos
Metilação de DNA , Idade Paterna , Animais , Epigênese Genética , Pai , Humanos , Masculino , Camundongos , Espermatozoides/metabolismo
4.
Neurosci Res ; 163: 63-67, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-32194144

RESUMO

Impaired social facilitation was reported in autism spectrum disorder (ASD) children. However, behavioral analysis methods of social facilitation for ASD model have not been reported. We developed a novel breeding home cage for social facilitation. Voluntary exercise of more social C57BL/6 J mice was significantly increased in the presence of observer mouse compared to that in the absence of observer mouse. In contrast, the presence of observer mouse did not affect voluntary exercise of less social BALB/cCrSlc mice. These suggest that BALB/cCrSlc mice, a mouse model of ASD, exhibited impaired social facilitation. Our method would provide novel clues for ASD pathophysiology.


Assuntos
Transtorno do Espectro Autista , Animais , Modelos Animais de Doenças , Camundongos , Camundongos Endogâmicos C57BL , Comportamento Social , Facilitação Social
5.
Nutrients ; 12(7)2020 Jul 09.
Artigo em Inglês | MEDLINE | ID: mdl-32659954

RESUMO

Obesity is characterized by massive adipose tissue accumulation and is associated with psychiatric disorders and cognitive impairment in human and animal models. However, it is unclear whether high-fat diet (HFD)-induced obesity presents a risk of psychiatric disorders and cognitive impairment. To examine this question, we conducted systematic behavioral analyses in C57BL/6J mice (male, 8-week-old) fed an HFD for 7 weeks. C57BL/6J mice fed an HFD showed significantly increased body weight, hyperlocomotion in the open-field test (OFT) and Y-maze test (YMZT), and impaired sucrose preference in the sucrose consumption test, compared to mice fed a normal diet. Neither body weight nor body weight gain was associated with any of the behavioral traits we examined. Working memory, as assessed by the YMZT, and anxiety-like behavior, as assessed by the elevated plus maze test (EPMT), were significantly correlated with mice fed an HFD, although these behavioral traits did not affect the entire group. These results suggest that HFD-induced obesity does not induce neuropsychiatric symptoms in C57BL/6J mice. Rather, HFD improved working memory in C57BL/6J mice with less anxiety, indicating that an HFD might be beneficial under limited conditions. Correlation analysis of individual traits is a useful tool to determine those conditions.


Assuntos
Ansiedade , Dieta Hiperlipídica , Aprendizagem em Labirinto , Memória de Curto Prazo , Anedonia , Animais , Comportamento Animal , Cognição , Teste de Labirinto em Cruz Elevado , Masculino , Camundongos Endogâmicos C57BL , Atividade Motora , Teste de Campo Aberto , Aumento de Peso
6.
Adv Exp Med Biol ; 1012: 75-81, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29956196

RESUMO

Sincethe theory of DOHaD has been thrown in the spotlight, most attention has focused on environmental effects of the uterus on developing embryos/fetuses. However, the ontogenesis traces back to gametogenesis. Compared to oogenesis, spermatogenesis goes through far more cell divisions and is therefore more prone to genetic variation and epigenetic alterations. This article will mainly discuss recent findings about the effects of the advanced paternal age on the next generation, in relation to the onset of psychiatric disorders such as autism spectrum disorder. We would like to advocate for further exploration on the DOHaD theory in a wider view.


Assuntos
Epigênese Genética/fisiologia , Transtornos do Neurodesenvolvimento/etiologia , Idade Paterna , Efeitos Tardios da Exposição Pré-Natal , Espermatozoides/metabolismo , Envelhecimento/genética , Envelhecimento/fisiologia , Transtorno do Espectro Autista/genética , Metilação de DNA/fisiologia , Pai , Feminino , Humanos , Masculino , Transtornos do Neurodesenvolvimento/genética , Gravidez , Efeitos Tardios da Exposição Pré-Natal/genética , Fatores de Risco , Espermatozoides/patologia
8.
PLoS One ; 12(11): e0186798, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29091920

RESUMO

The understanding of individual diversity and its link to brain functions is a fundamental issue in neurobiology. Studies in mice have mainly focused on the investigation of behavior traits in adulthood, whereas longitudinal analyses are largely uninvestigated. Here we have conducted systematic behavior tests in individual mice (C57BL6/J, male), comparing phenotypes at early postnatal stages and in adulthood. Each animal showed different scores in individual behavior tests. However, we observed an inverse correlation between repetitive behavior in the Morris water maze test and sociability in the 3-chamber social interaction test; an increase in repetitive behaviors was associated with poor sociability. In longitudinal analyses, the emission of ultrasonic vocalization during maternal separation at postnatal day 6 in pups was correlated positively with sociability and negatively with spatial memory. Our results show a possibility that individual differences in communication between pups and their mother in infancy is a predictive indicator for sociability and cognitive performance as an adult.


Assuntos
Comunicação Animal , Comportamento Animal , Memória , Animais , Camundongos , Camundongos Endogâmicos C57BL
9.
PLoS One ; 11(11): e0166665, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27855195

RESUMO

Neurodevelopmental disorders such as autism spectrum disorder (ASD) and attention deficit and hyperactivity disorder (ADHD) have increased over the last few decades. These neurodevelopmental disorders are characterized by a complex etiology, which involves multiple genes and gene-environmental interactions. Various genes that control specific properties of neural development exert pivotal roles in the occurrence and severity of phenotypes associated with neurodevelopmental disorders. Moreover, paternal aging has been reported as one of the factors that contribute to the risk of ASD and ADHD. Here we report, for the first time, that paternal aging has profound effects on the onset of behavioral abnormalities in mice carrying a mutation of Pax6, a gene with neurodevelopmental regulatory functions. We adopted an in vitro fertilization approach to restrict the influence of additional factors. Comprehensive behavioral analyses were performed in Sey/+ mice (i.e., Pax6 mutant heterozygotes) born from in vitro fertilization of sperm taken from young or aged Sey/+ fathers. No body weight changes were found in the four groups, i.e., Sey/+ and wild type (WT) mice born to young or aged father. However, we found important differences in maternal separation-induced ultrasonic vocalizations of Sey/+ mice born from young father and in the level of hyperactivity of Sey/+ mice born from aged fathers in the open-field test, respectively, compared to WT littermates. Phenotypes of anxiety were observed in both genotypes born from aged fathers compared with those born from young fathers. No significant difference was found in social behavior and sensorimotor gating among the four groups. These results indicate that mice with a single genetic risk factor can develop different phenotypes depending on the paternal age. Our study advocates for serious considerations on the role of paternal aging in breeding strategies for animal studies.


Assuntos
Envelhecimento/patologia , Comportamento Animal , Interação Gene-Ambiente , Transtornos do Neurodesenvolvimento/genética , Transtornos do Neurodesenvolvimento/patologia , Fator de Transcrição PAX6/genética , Afeto , Animais , Ansiedade/fisiopatologia , Medo , Feminino , Locomoção , Masculino , Memória , Camundongos Endogâmicos C57BL , Camundongos Mutantes , Fator de Transcrição PAX6/metabolismo , Idade Paterna , Fenótipo , Filtro Sensorial , Comportamento Social , Ultrassom , Vocalização Animal
10.
Tohoku J Exp Med ; 240(1): 31-7, 2016 09.
Artigo em Inglês | MEDLINE | ID: mdl-27558477

RESUMO

Omega-6 (n-6) and omega-3 (n-3) polyunsaturated fatty acids (PUFAs) are essential nutrients for normal brain development. The principal dietary n-6 and n-3 PUFAs are linoleic acid (LA) and α-linolenic acid (ALA), respectively, We have previously shown that maternal dietary imbalance between these PUFAs, i.e., rich in LA and poor in ALA, affected brain development and increased anxiety-related behavior in the mouse offspring. Here we further addressed sex difference in anxiety-related behavior in the offspring exposed to maternal LA:ALA imbalance. We fed pregnant mice a LA excess/ALA deficient (LA(ex)/ALA(def)) diet, and raised their offspring on a well-balanced LA:ALA diet from an early lactation period. When the offspring were grown to adulthood, they were subjected to behavioral and biochemical analyses. We found that both male and female offspring exposed to the LA(ex)/ALA(def) diet showed increased anxiety-related behavior compared to those exposed to the control diet, which was differently observed between the sexes. The female offspring also exhibited hyperactivity by maternal intake of the LA(ex)/ALA(def) diet. On the other hand, abnormal depressive behavior was undetected in both sexes. We also found that the ratio of n-6 to n-3 PUFAs in the brain was unaffected regardless of maternal diet or offspring's sex. Since the n-6/n-3 ratio is known to influence emotional behavior, it is reasonable to assume that LA:ALA imbalance exposed during brain development is the key for causing enhanced anxiety in adulthood. The present study indicates that maternal dietary imbalance between LA and ALA increases offspring's anxiety-related behavior with a sex-dependent manner.


Assuntos
Ansiedade/fisiopatologia , Ácido Linoleico/metabolismo , Fenômenos Fisiológicos da Nutrição Materna , Caracteres Sexuais , Ácido alfa-Linolênico/metabolismo , Animais , Ansiedade/complicações , Peso Corporal , Encéfalo/metabolismo , Depressão/complicações , Depressão/fisiopatologia , Dieta , Comportamento Alimentar , Feminino , Masculino , Aprendizagem em Labirinto , Camundongos Endogâmicos C57BL , Atividade Motora
11.
Stem Cells ; 34(2): 470-82, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26580686

RESUMO

Omega-6 (n-6) and omega-3 (n-3) polyunsaturated fatty acids (PUFAs) are essential nutrients. Although several studies have suggested that a balanced dietary n-6:n-3 ratio is essential for brain development, the underlying cellular and molecular mechanism is poorly understood. Here, we found that feeding pregnant mice an n-6 excess/n-3 deficient diet, which reflects modern human diets, impairsed neocortical neurogenesis in the offspring. This impaired neurodevelopment occurs through a precocious fate transition of neural stem cells from the neurogenic to gliogenic lineage. A comprehensive mediator lipidomics screen revealed key mediators, epoxy metabolites, which were confirmed functionally using a neurosphere assay. Importantly, although the offspring were raised on a well-balanced n-6:n-3 diet, they exhibited increased anxiety-related behavior in adulthood. These findings provide compelling evidence that excess maternal consumption of n-6 PUFAs combined with insufficient intake of n-3 PUFAs causes abnormal brain development that can have long-lasting effects on the offspring's mental state.


Assuntos
Ácidos Graxos Ômega-3 , Ácidos Graxos Ômega-6/deficiência , Exposição Materna/efeitos adversos , Neocórtex/crescimento & desenvolvimento , Células-Tronco Neurais/metabolismo , Neurogênese , Animais , Ácidos Graxos Ômega-6/metabolismo , Feminino , Camundongos , Neocórtex/patologia , Células-Tronco Neurais/patologia , Gravidez
12.
J Anat ; 227(1): 1-9, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-26032914

RESUMO

Spermatogenesis is a series of complex processes to generate mature sperm, and various molecules play crucial roles in regulating these processes. Previous studies imply a possibility that a transcriptional factor Pax6, a key player of brain and sensory organ development, could be involved in spermatogenesis, but neither expression nor function of Pax6 in the adult testis has been examined yet. In the present study, we described for the first time Pax6 expression dynamics in the adult mouse testis. Using cell-type-specific markers, the expression of Pax6 was detected in 67.0% of promyelocytic leukemia zinc finger (Plzf)-positive type A spermatogonia. The expression of Pax6 was also observed in p63-positive spermatocytes and round spermatids. We did not detect any expression of Pax6 in Sox9-positive Sertoli cells or in elongated spermatids and mature sperm. High-resolution analyses revealed that Pax6 formed a single dot-like structure during mid-phase of the pachytene spermatocyte. This dot-like structure co-localized with γH2A.X demarcating XY body, a domain in which X and Y chromosomes are silenced and compartmentalized. These results may suggest a novel role of Pax6 in spermatogenesis.


Assuntos
Proteínas do Olho/metabolismo , Proteínas de Homeodomínio/metabolismo , Fatores de Transcrição Box Pareados/metabolismo , Proteínas Repressoras/metabolismo , Espermatogênese/fisiologia , Testículo/metabolismo , Animais , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Fator de Transcrição PAX6 , Células de Sertoli/metabolismo , Espermatócitos/metabolismo , Espermatogônias/metabolismo , Testículo/citologia
13.
J Neurosci ; 33(15): 6691-704, 2013 Apr 10.
Artigo em Inglês | MEDLINE | ID: mdl-23575865

RESUMO

Developmental perturbations during adolescence have been hypothesized to be a risk factor for the onset of several neuropsychiatric diseases. However the physiological alterations that result from such insults are incompletely understood. We investigated whether a defined perturbation during adolescence affected hippocampus-dependent sensorimotor gating functions, a proposed endophenotype in several psychiatric diseases, most notably schizophrenia. The developmental perturbation was induced during adolescence in mice using an antimitotic agent, methylazoxymethanol acetate (MAM), during postnatal weeks (PW) 4-6. MAM-treated mice showed a decrease in hippocampal neurogenesis immediately after treatment, which was restored by PW10 in adulthood. However, the mice treated with MAM during adolescent stages exhibited a persistent sensorimotor gating deficiency and a reduction in prepulse inhibition-related activation of hippocampal and prefrontal neurons in adulthood. Cellular analyses found a reduction of GABAergic inhibitory neurons and abnormal dendritic morphology of immature neurons in the dentate gyrus (DG). Interestingly, bilateral infusion of muscimol, a GABAA receptor agonist, into the DG region reversed the prepulse inhibition abnormality in MAM-treated mice. Furthermore, the behavioral deficits together with the decrease in the number of GABAergic neurons in this MAM model were rescued by exposure to an enriched environment during a defined critical adolescent period. These observations suggest a possible role for GABAergic interneurons in the DG during adolescence. This role may be related to the establishment of neural circuitry required for sensorimotor gating. It is plausible that changes in neurogenesis during this window may affect the survival of GABAergic interneurons, although this link needs to be causally addressed.


Assuntos
Período Crítico Psicológico , Giro Denteado/fisiologia , Neurônios GABAérgicos/fisiologia , Interneurônios/fisiologia , Acetato de Metilazoximetanol/toxicidade , Filtro Sensorial/fisiologia , Fatores Etários , Animais , Comportamento Animal/efeitos dos fármacos , Comportamento Animal/fisiologia , Dendritos , Giro Denteado/metabolismo , Modelos Animais de Doenças , Proteínas do Domínio Duplacortina , Quinases Semelhantes a Duplacortina , Interações Medicamentosas , Meio Ambiente , Agonistas GABAérgicos/administração & dosagem , Agonistas GABAérgicos/farmacologia , Neurônios GABAérgicos/metabolismo , Inibição Psicológica , Interneurônios/citologia , Interneurônios/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Microinjeções , Proteínas Associadas aos Microtúbulos/metabolismo , Muscimol/administração & dosagem , Muscimol/farmacologia , Muscimol/uso terapêutico , Neurogênese/efeitos dos fármacos , Neuropeptídeos/metabolismo , Córtex Pré-Frontal/fisiologia , Proteínas Serina-Treonina Quinases , Esquizofrenia/induzido quimicamente , Esquizofrenia/tratamento farmacológico , Esquizofrenia/fisiopatologia
14.
Neurosci Res ; 75(2): 112-20, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23228673

RESUMO

Horizontal basal cells (HBCs) have garnered attention as tissue stem cells of the olfactory epithelium (OE); however, these cells' exact lineage and their contributions to OE regeneration remain unknown. Neural crest-derived cells (NCDCs) have been shown to possess stem cell properties and to participate in the normal development of the OE. However, the contributions of NCDCs to both normal and regenerating adult OE remain unclear. In this study, we investigated the contribution of NCDCs to the OE, focusing particularly on HBCs. Using immunohistochemistry, we observed the OE of P0-Cre/EGFP mice expressing EGFP-tagged NCDCs at several stages of normal development along with regenerated OE following methimazole treatment. We observed EGFP expression in the HBCs, sustentacular cells (SUSs), Bowman's glands, olfactory receptor neurons (ORNs), and olfactory ensheathing cells of 6-week-old mice. No ectopic Cre expression was identified. Although HBCs at late embryonic stages were placode-derived (i.e., EGFP-negative), we found that EGFP+ HBCs alternatively increased with the decrease of placode-derived HBCs during maturation. In regenerated OE, the percentages of neural crest-derived ORNs and SUSs significantly increased compared with normal OE. These results suggest that NCDCs contribute greatly to the adult HBC population and that they are important for the maintenance of the OE.


Assuntos
Crista Neural/citologia , Células-Tronco Neurais/citologia , Mucosa Olfatória/citologia , Neurônios Receptores Olfatórios/metabolismo , Envelhecimento , Animais , Diferenciação Celular/fisiologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Crista Neural/metabolismo , Células-Tronco Neurais/metabolismo , Mucosa Olfatória/metabolismo , Neurônios Receptores Olfatórios/citologia
15.
Brain Nerve ; 62(12): 1315-22, 2010 Dec.
Artigo em Japonês | MEDLINE | ID: mdl-21139184

RESUMO

Postnatal neurogenesis has been observed in two brain regions: the subventricular zone (SVZ) of the lateral ventricle and the subgranular zone (SGZ) of the dentate gyrus in the hippocampus, among vertebrates including human. Accumulating evidence has indicated the molecular mechanisms commonly underlying embryonic and adult neurogenesis. Genetic factors essential for neural development, i.e., Pax6, Fabp7, Sox2, Wnt3, Notch1, etc., are also expressed in adult neurogenic regions. Adult neurogenesis, however, is distinct from embryonic neurogenesis in that the former is activity dependent; environmental stimulation modulates the entire processes of adult neurogenesis. In the hippocampus, physical exercise and cognitive stimuli robustly increase the proliferation of precursor cells, whereas physical/psychosocial stress decreases the proliferation of newborn neurons. Thus, adult neurogenesis is intriguingly regulated by several genetic and environmental factors. Reduction in hippocampal neurogenesis during the infantile and adult stages has been observed in some animal models of mental illness such as schizophrenia and major depression, implicating that postnatal neurogenesis may contribute to a part of the symptoms of mental illness. In this review, we describe the molecular mechanisms and functional significance of postnatal neurogenesis.


Assuntos
Encéfalo/crescimento & desenvolvimento , Proteínas de Transporte/fisiologia , Proteínas do Olho/fisiologia , Proteínas de Homeodomínio/fisiologia , Neurônios/citologia , Fatores de Transcrição Box Pareados/fisiologia , Proteínas Repressoras/fisiologia , Proteínas Supressoras de Tumor/fisiologia , Animais , Proliferação de Células , Transtorno Depressivo Maior/etiologia , Meio Ambiente , Proteína 7 de Ligação a Ácidos Graxos , Humanos , Recém-Nascido , Fator de Transcrição PAX6 , Receptor Notch1/fisiologia , Fatores de Transcrição SOXB1/fisiologia , Esquizofrenia/etiologia , Estresse Psicológico/fisiopatologia , Proteínas Wnt/fisiologia , Proteína Wnt3
16.
Stem Cells ; 28(5): 974-83, 2010 May.
Artigo em Inglês | MEDLINE | ID: mdl-20474079

RESUMO

Neurogenesis occurs throughout the life in the mammalian brain. The hippocampal dentate gyrus (DG) is one of the major regions of the adult neurogenesis, where neural stem/progenitor cells continuously generate new granule neurons, although molecular mechanisms underlying generation and maintenance of newly born neurons are still elusive. Here we show that ephrin-A5, a ligand for Eph receptor tyrosine kinases, plays multiple roles in both neurogenesis and vascular formation in the adult hippocampus. In mice lacking ephrin-A5 function, cell proliferation and survival of newborn neurons were severely reduced in the hippocampus DG. Furthermore, ephrin-A5-deficient mice exhibited altered distribution of EphA4 receptor in the vascular endothelial cells and increased narrower capillaries in the hippocampus DG. EphA/ephrin-A signaling thus plays crucial roles in the establishment and/or maintenance of the brain vascular system, as an essential constituent of the adult neurogenic niche.


Assuntos
Células-Tronco Adultas/metabolismo , Efrina-A5/genética , Hipocampo/metabolismo , Neovascularização Patológica/metabolismo , Neovascularização Patológica/patologia , Neurogênese/fisiologia , Células-Tronco Adultas/citologia , Envelhecimento/genética , Animais , Proliferação de Células , Sobrevivência Celular/genética , Regulação para Baixo/genética , Endotélio Vascular/metabolismo , Endotélio Vascular/patologia , Hipocampo/citologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Neovascularização Patológica/genética , Receptores da Família Eph/genética , Receptores da Família Eph/metabolismo
17.
Hum Mol Genet ; 19(1): 79-89, 2010 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-19825845

RESUMO

Mutations in the human NOTCH3 gene cause cerebral autosomal-dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL), but the pathogenic mechanisms of the disorder remain unclear. We investigated the cytotoxic properties of mutant Notch3 using stable cell lines with inducible expression of either wild-type or two mutants p.R133C and p.C185R. We found that both mutants of Notch3 were prone to aggregation and retained in the endoplasmic reticulum (ER). The turnover rates of the mutated Notch3 proteins were strikingly slow, with half-lives greater than 6 days, whereas wild-type Notch3 was rapidly degraded, with a half-life of 0.7 days. The expression of mutant Notch3 also impaired cell proliferation compared with wild-type Notch3. In addition, cell lines expressing mutant Notch3 were more sensitive to proteasome inhibition resulting in cell death. These findings suggest that prolonged retention of mutant Notch3 aggregates in the ER decreases cell growth and increases sensitivity to other stresses. It is also possible that the aggregate-prone property of mutant Notch3 contributes to a pathogenic mechanism underlying CADASIL.


Assuntos
Retículo Endoplasmático/patologia , Mutação/genética , Receptores Notch/química , Receptores Notch/genética , Western Blotting , Morte Celular/efeitos dos fármacos , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Retículo Endoplasmático/efeitos dos fármacos , Retículo Endoplasmático/metabolismo , Humanos , Imuno-Histoquímica , Cinética , Leupeptinas/farmacologia , Chaperonas Moleculares/metabolismo , Proteínas Mutantes/metabolismo , Complexo de Endopeptidases do Proteassoma/metabolismo , Processamento de Proteína Pós-Traducional/efeitos dos fármacos , Estrutura Quaternária de Proteína , Transporte Proteico/efeitos dos fármacos , Receptor Notch3 , Receptores Notch/metabolismo , Frações Subcelulares/efeitos dos fármacos , Frações Subcelulares/metabolismo
18.
Biosci Biotechnol Biochem ; 73(2): 311-5, 2009 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-19202296

RESUMO

Mild oxidative stress is known to induce premature senescence, termed stress-induced premature senescence (SIPS), in normal human diploid cells. We investigated to determine whether mild oxidative stress would trigger SIPS in a human tumor cell line, human lung adenocarcinoma A549. The results showed that sublethal concentrations of H(2)O(2) induced SIPS in A549 cells and consequently attenuated, but did not completely eliminate, the tumorigenicity of these cells. We next investigated the reasons for this incomplete impairment of tumorigenicity in A549 cells in SIPS. The results suggested that H(2)O(2)-treated A549 cells are composed of a heterogeneous cell population: one is sensitive to H(2)O(2), and the other is resistant or undergoes reversal; the latter reverted to their original tumorigenic form. The molecular mechanisms determining the cellular fate of tumor cells in SIPS should be identified in order to make use of SIPS and oncogene-induced senescence in tumor cells as methods of tumor suppression.


Assuntos
Transformação Celular Neoplásica/efeitos dos fármacos , Senescência Celular/efeitos dos fármacos , Peróxido de Hidrogênio/farmacologia , Peróxido de Hidrogênio/uso terapêutico , Neoplasias/tratamento farmacológico , Neoplasias/patologia , Animais , Linhagem Celular Tumoral , Feminino , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Neoplasias/metabolismo , Estresse Oxidativo/efeitos dos fármacos
19.
Biosci Biotechnol Biochem ; 72(6): 1638-41, 2008 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-18540079

RESUMO

We constructed a novel cancer-specific regulatable adenoviral expression system comprising two vectors: one expressing rtTA, a reverse tetracycline transactivator regulated by the human telomerase reverse transcriptase (hTERT) gene promoter, the other expressing the target gene regulated by the tetracycline response element (TRE). rtTA transactivates target gene expression in the presence of doxycycline. Using these vectors, we constructed an adenoviral expression system, the Tel-On system, which enables highly efficient target gene transduction. This system enabled efficient and regulatable cancer-specific gene expression, and can be used in targeted cancer gene therapy.


Assuntos
Adenoviridae/genética , Expressão Gênica , Neoplasias/metabolismo , Regiões Promotoras Genéticas/genética , Telomerase/genética , Linhagem Celular Tumoral , Vetores Genéticos/genética , Humanos , Neoplasias/genética
20.
Biochem Biophys Res Commun ; 371(4): 747-51, 2008 Jul 11.
Artigo em Inglês | MEDLINE | ID: mdl-18471997

RESUMO

Human E-selectin, an endothelial adhesion molecule, is induced in the brain arteries by cerebral ischemia and participates in the infiltration of leukocytes that cause inflammatory reaction leading to brain damage. To prevent leukocyte infiltration in the brain, we designed gene therapeutic constructs to suppress E-selectin expression. The constructs were composed of microRNAs (miR-E1 and miR-E2) complementary to the human E-selectin cDNA, which were directed by a minimum cis-element of the human E-selectin promoter. Transfection in human aorta endothelial cells (HAECs) with these constructs revealed that the E-selectin promoter was sufficiently activated in response to tumor necrosis factor-alpha (TNF-alpha), and miR-E1 and miR-E2 could suppress E-selectin expression resulting in the significant inhibition of leukocyte adhesion. These results suggested that the combination of the E-selectin promoter and microRNAs could allow the restricted expression of transgenes in activated endothelial cells and diminish leukocyte recruitment.


Assuntos
Selectina E/genética , Endotélio Vascular/imunologia , Inflamação/imunologia , Leucócitos/imunologia , MicroRNAs/genética , Transgenes , Aorta/citologia , Adesão Celular/genética , Células Cultivadas , Endotélio Vascular/citologia , Expressão Gênica , Humanos , Regiões Promotoras Genéticas , Transfecção , Fator de Necrose Tumoral alfa/metabolismo
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