RESUMO
Social interaction is a fundamental behavior in all animal species, but the developmental timing of the social neural circuit formation and the cellular and molecular mechanisms governing its formation are poorly understood. We generated a mouse model with mutations in two Disheveled genes, Dvl1 and Dvl3, that displays adult social and repetitive behavioral abnormalities associated with transient embryonic brain enlargement during deep layer cortical neuron formation. These phenotypes were mediated by the embryonic expansion of basal neural progenitor cells (NPCs) via deregulation of a ß-catenin/Brn2/Tbr2 transcriptional cascade. Transient pharmacological activation of the canonical Wnt pathway during this period of early corticogenesis rescued the ß-catenin/Brn2/Tbr2 transcriptional cascade and the embryonic brain phenotypes. Remarkably, this embryonic treatment prevented adult behavioral deficits and partially rescued abnormal brain structure in Dvl mutant mice. Our findings define a mechanism that links fetal brain development and adult behavior, demonstrating a fetal origin for social and repetitive behavior deficits seen in disorders such as autism.
Assuntos
Transtorno de Movimento Estereotipado/genética , Transtorno de Movimento Estereotipado/fisiopatologia , Proteínas Adaptadoras de Transdução de Sinal/genética , Animais , Comportamento Animal , Encéfalo/embriologia , Encéfalo/metabolismo , Encéfalo/fisiologia , Proteínas Desgrenhadas/genética , Proteínas Desgrenhadas/metabolismo , Humanos , Camundongos , Proteínas do Tecido Nervoso/metabolismo , Proteínas do Tecido Nervoso/fisiologia , Células-Tronco Neurais/metabolismo , Neurônios/metabolismo , Fatores do Domínio POU/metabolismo , Fatores do Domínio POU/fisiologia , Fosfoproteínas/genética , Transdução de Sinais/fisiologia , Comportamento Estereotipado/fisiologia , Proteínas com Domínio T/metabolismo , Proteínas com Domínio T/fisiologia , Proteínas Wnt/metabolismo , Via de Sinalização Wnt/genética , Via de Sinalização Wnt/fisiologia , beta Catenina/metabolismo , beta Catenina/fisiologiaRESUMO
Clubfoot is a common birth defect characterized by inward posturing and rigid downward displacement of one or both feet. The etiology of syndromic forms of clubfoot is varied and the causes of isolated clubfoot are not well understood. A microduplication of 2.2 Mb on chromosome 17q23.1q23.2 which includes T-box 4 (TBX4), a hindlimb-specific gene, and 16 other genes was recently identified in 3 of 66 families reported as nonsyndromic clubfoot, but additional non-foot malformations place them in the syndromic clubfoot category. Our study assesses whether variation in or around TBX4 contributes to nonsyndromic clubfoot. To determine whether this microduplication was a common cause of nonsyndromic clubfoot, 605 probands (from 148 multiplex and 457 simplex families) with nonsyndromic clubfoot were evaluated by copy number and oligonucleotide array CGH testing modalities. Only one multiplex family (0.68%) that had 16 with clubfoot and 9 with other foot anomalies, had a 350 kb microduplication, which included the complete duplication of TBX4 and NACA2 and partial duplication of BRIP1. The microduplication was transmitted in an autosomal dominant pattern and all with the microduplication had a range of phenotypes from short wide feet and toes to bilateral clubfoot. Minimal evidence was found for an association between TBX4 and clubfoot and no pathogenic sequence variants were identified in the two known TBX4 hindlimb enhancer elements. Altogether, these results demonstrate that variation in and around the TBX4 gene and the 17q23.1q23.2 microduplication are not a frequent cause of this common orthopedic birth defect and narrows the 17q23.1q23.2 nonsyndromic clubfoot-associated region.
Assuntos
Duplicação Cromossômica , Cromossomos Humanos Par 17 , Pé Torto Equinovaro/genética , Proteínas com Domínio T/genética , Alelos , Sequência de Bases , Variações do Número de Cópias de DNA , Elementos Facilitadores Genéticos , Feminino , Humanos , Masculino , Linhagem , Fenótipo , Polimorfismo de Nucleotídeo ÚnicoRESUMO
The molecular basis for autosomal dominant progressive nonsyndromic hearing loss in an Israeli Jewish family, Family H, has been determined. Linkage analysis placed this deafness locus, DFNA15, on chromosome 5q31. The human homolog of mouse Pou4f3, a member of the POU-domain family of transcription factors whose targeted inactivation causes profound deafness in mice, was physically mapped to the 25-centimorgan DFNA15-linked region. An 8-base pair deletion in the POU homeodomain of human POU4F3 was identified in Family H. A truncated protein presumably impairs high-affinity binding of this transcription factor in a dominant negative fashion, leading to progressive hearing loss.
Assuntos
Surdez/genética , Perda Auditiva Neurossensorial/genética , Proteínas de Homeodomínio/genética , Fatores de Transcrição/genética , Adulto , Animais , Diferenciação Celular , Mapeamento Cromossômico , Cromossomos Humanos Par 5/genética , Feminino , Expressão Gênica , Ligação Genética , Células Ciliadas Auditivas/citologia , Células Ciliadas Auditivas/fisiologia , Proteínas de Homeodomínio/metabolismo , Humanos , Israel , Judeus/genética , Masculino , Camundongos , Pessoa de Meia-Idade , Dados de Sequência Molecular , Linhagem , Reação em Cadeia da Polimerase , Estrutura Secundária de Proteína , Deleção de Sequência , Fator de Transcrição Brn-3C , Fatores de Transcrição/metabolismo , Fatores de Transcrição/fisiologiaRESUMO
Mutations in myosin VI (Myo6) cause deafness and vestibular dysfunction in Snell's waltzer mice. Mutations in two other unconventional myosins cause deafness in both humans and mice, making myosin VI an attractive candidate for human deafness. In this report, we refined the map position of human myosin VI (MYO6) by radiation hybrid mapping and characterized the genomic structure of myosin VI. Human myosin VI is composed of 32 coding exons, spanning a genomic region of approximately 70 kb. Exon 30, containing a putative CKII site, was found to be alternatively spliced and appears only in fetal and adult human brain. D6S280 and D6S284 flank the myosin VI gene and were used to screen hearing impaired sib pairs for concordance with the polymorphic markers. No disease-associated mutations were identified in twenty-five families screened for myosin VI mutations by SSCP analysis. Three coding single nucleotide polymorphisms (cSNPs) were identified in myosin VI that did not alter the amino acid sequence. Myosin VI mutations may be rare in the human deaf population or alternatively, may be found in a population not yet examined. The determination of the MYO6 genomic structure will enable screening of individuals with non-syndromic deafness, Usher's syndrome, or retinopathies associated with human chromosome 6q for mutations in this unconventional myosin.
Assuntos
Genes/genética , Cadeias Pesadas de Miosina/genética , Adulto , Sequência de Bases , Encéfalo/embriologia , Encéfalo/metabolismo , DNA/química , DNA/genética , Análise Mutacional de DNA , Surdez/genética , Éxons , Saúde da Família , Feto , Frequência do Gene , Humanos , Íntrons , Mutação , Mutação Puntual , Polimorfismo de Nucleotídeo Único , Polimorfismo Conformacional de Fita SimplesRESUMO
We have shown here several examples of how hearing and vestibular impaired mouse mutants are generated and the insight that they provide in the study of auditory and vestibular function. These types of genetic studies may also lead to the identification of disease-susceptibility genes, perhaps the most critical element in presbyacusis (age-related hearing loss). Some individuals may be more prone to hearing loss with increasing age or upon exposure to severe noise, and susceptibility genes may be involved. Different inbred mice show a variety of age-related and noise-induced hearing loss that varies between normal hearing and severe deafness throughout their life span /27/. Genetic diversity between inbred mouse strains has been shown to be a powerful tool for the discovery of modifier genes. Already two studies have found regions in which modifier genes for deafness may reside /28-29/. Future studies will hopefully lead to the identification of genes that modify hearing loss and will help us understand the variability that exists in human hearing, a crucial component in developing successful treatment strategies. The first human non-syndromic deafness-causing gene was identified in 1995, and since then, additional genes have been discovered. Much of the credit for this boom is due to deaf and vestibular mouse mutants. Their study has led to great insight regarding the development and function of the mammalian inner ear, and correlations with human deafness can now be made since mutations in the same genes have been found in these two mammals. As deafness is the most common form of sensory impairment and affects individuals of all ages, elucidating the function of the auditory and vestibular systems through genetic approaches is essential in improving and designing effective treatments for hearing loss.
Assuntos
Proteínas de Ligação a DNA , Surdez/etiologia , Modelos Animais de Doenças , Proteínas do Tecido Nervoso , Animais , Conexina 26 , Conexinas/genética , Surdez/genética , Humanos , Camundongos , Camundongos Knockout , Camundongos Mutantes , Miosinas/genética , Fatores do Domínio POU , Fatores de Transcrição/genéticaRESUMO
A mathematical programming model is proposed to select an optimal cooperation policy between autonomous service networks for dispatching purposes. In addition to traditional characteristics such as network topology and station location, this model takes into account 'political' constraints on minimum response-time in certain subzones. Such constraints are translated into performance requirements, which are imposed on the cooperation policy. Testing the model under different assumptions can be useful for analysing various cooperation policies. The paper formulates a mathematical programming model, derives example policies for various circumstances, and tests the sensitivity of the resultant policies to some parameters, such as the penalty for not providing service, and distances between adjacent networks. The paper suggests also a less constrained approach, which entails a linear programming model. A comparison between the two approaches is presented.
Assuntos
Técnicas de Apoio para a Decisão , Serviços Médicos de Emergência/organização & administração , Modelos Teóricos , Formulação de Políticas , Comportamento Cooperativo , Fatores de TempoRESUMO
The autism susceptibility candidate 2 gene (AUTS2) has been associated with multiple neurological diseases including autism spectrum disorders (ASDs). Previous studies showed that AUTS2 has an important neurodevelopmental function and is a suspected master regulator of genes implicated in ASD-related pathways. However, the regulatory role and targets of Auts2 are not well known. Here, by using ChIP-seq (chromatin immunoprecipitation followed by deep sequencing) and RNA-seq on mouse embryonic day 16.5 forebrains, we elucidated the gene regulatory networks of Auts2. We find that the majority of promoters bound by Auts2 belong to genes highly expressed in the developing forebrain, suggesting that Auts2 is involved in transcriptional activation. Auts2 non-promoter-bound regions significantly overlap developing brain-associated enhancer marks and are located near genes involved in neurodevelopment. Auts2-marked sequences are enriched for binding site motifs of neurodevelopmental transcription factors, including Pitx3 and TCF3. In addition, we characterized two functional brain enhancers marked by Auts2 near NRXN1 and ATP2B2, both ASD-implicated genes. Our results implicate Auts2 as an active regulator of important neurodevelopmental genes and pathways and identify novel genomic regions that could be associated with ASD and other neurodevelopmental diseases.
Assuntos
Transtornos Globais do Desenvolvimento Infantil/genética , Predisposição Genética para Doença/genética , Estudo de Associação Genômica Ampla , Proteínas Nucleares/genética , Animais , Proteínas de Ligação ao Cálcio , Proteínas do Citoesqueleto , Feminino , Regulação da Expressão Gênica/genética , Camundongos , Camundongos Endogâmicos , Moléculas de Adesão de Célula Nervosa/genética , ATPases Transportadoras de Cálcio da Membrana Plasmática/genética , Gravidez , Prosencéfalo/embriologia , Fatores de Transcrição/genética , Ativação Transcricional/genéticaRESUMO
Little is known about how genetic variations in enhancers influence drug response. In this study, we investigated whether nucleotide variations in enhancers that regulate drug transporters can alter their expression levels. Using comparative genomics and liver-specific transcription factor binding site (TFBS) analyses, we identified evolutionary conserved regions (ECRs) surrounding nine liver membrane transporters that interact with commonly used pharmaceuticals. The top 50 ECRs were screened for enhancer activity in vivo, of which five--located around ABCB11, SLC10A1, SLCO1B1, SLCO1A2, and SLC47A1--exhibited significant enhancer activity. Common variants identified in a large ethnically diverse cohort (n = 272) were assayed for differential enhancer activity, and three variants were found to have significant effects on reporter activity as compared with the reference allele. In addition, one variant was associated with reduced SLCO1A2 mRNA expression levels in human liver tissues, and another was associated with increased methotrexate (MTX) clearance in patients. This work provides a general model for the rapid characterization of liver enhancers and identifies associations between enhancer variants and drug response.
Assuntos
Transportadores de Cassetes de Ligação de ATP/metabolismo , Metotrexato/farmacocinética , Transportadores de Ânions Orgânicos/metabolismo , Proteínas de Transporte de Cátions Orgânicos/metabolismo , Preparações Farmacêuticas/metabolismo , Membro 11 da Subfamília B de Transportadores de Cassetes de Ligação de ATP , Transportadores de Cassetes de Ligação de ATP/genética , Alelos , Animais , Sítios de Ligação , Transporte Biológico , Sequência Conservada , Feminino , Regulação da Expressão Gênica , Variação Genética , Genômica/métodos , Humanos , Fígado/metabolismo , Masculino , Camundongos , Transportadores de Ânions Orgânicos/genética , Proteínas de Transporte de Cátions Orgânicos/genética , Polimorfismo de Nucleotídeo Único , RNA Mensageiro/metabolismo , Grupos Raciais/genética , Fatores de TranscriçãoRESUMO
Within the mammalian inner ear there are six separate sensory regions that subserve the functions of hearing and balance, although how these sensory regions become specified remains unknown. Each sensory region is populated by two cell types, the mechanosensory hair cell and the supporting cell, which are arranged in a mosaic in which each hair cell is surrounded by supporting cells. The proposed mechanism for creating the sensory mosaic is lateral inhibition mediated by the Notch signaling pathway. However, one of the Notch ligands, Jagged1 (Jag1), does not show an expression pattern wholly consistent with a role in lateral inhibition, as it marks the sensory patches from very early in their development--presumably long before cells make their final fate decisions. It has been proposed that Jag1 has a role in specifying sensory versus nonsensory epithelium within the ear [Adam, J., Myat, A., Roux, I. L., Eddison, M., Henrique, D., Ish-Horowicz, D. & Lewis, J. (1998) Development (Cambridge, U.K.) 125, 4645--4654]. Here we provide experimental evidence that Notch signaling may be involved in specifying sensory regions by showing that a dominant mouse mutant headturner (Htu) contains a missense mutation in the Jag1 gene and displays missing posterior and sometimes anterior ampullae, structures that house the sensory cristae. Htu/+ mutants also demonstrate a significant reduction in the numbers of outer hair cells in the organ of Corti. Because lateral inhibition mediated by Notch predicts that disruptions in this pathway would lead to an increase in hair cells, we believe these data indicate an earlier role for Notch within the inner ear.
Assuntos
Orelha Interna/crescimento & desenvolvimento , Proteínas de Membrana/metabolismo , Proteínas/fisiologia , Sequência de Aminoácidos , Animais , Proteínas de Ligação ao Cálcio , Mapeamento Cromossômico , Cromossomos , Orelha Interna/anormalidades , Homozigoto , Humanos , Peptídeos e Proteínas de Sinalização Intercelular , Proteína Jagged-1 , Camundongos , Camundongos Endogâmicos C3H , Camundongos Endogâmicos C57BL , Dados de Sequência Molecular , Mutação de Sentido Incorreto , Fenótipo , Ratos , Receptores Notch , Análise de Sequência de DNA , Proteínas Serrate-JaggedRESUMO
Mutations in the unconventional myosin VI gene, Myo6, are associated with deafness and vestibular dysfunction in the Snell's waltzer (sv) mouse. The corresponding human gene, MYO6, is located on chromosome 6q13. We describe the mapping of a new deafness locus, DFNA22, on chromosome 6q13 in a family affected by a nonsyndromic dominant form of deafness (NSAD), and the subsequent identification of a missense mutation in the MYO6 gene in all members of the family with hearing loss.