RESUMO
We recently observed a reliable phenotypic difference in the inflammatory pain sensitivity of a congenic mouse strain compared to its background strain. By constructing and testing subcongenic strains combined with gene-expression assays, we provide evidence for the candidacy of the Yy1 gene - encoding the ubiquitously expressed and multifunctional Yin Yang 1 transcription factor - as responsible. To confirm this hypothesis, we used a Cre/lox strategy to produce mutant mice in which Yy1 expression was ablated in Nav 1.8-positive neurons of the dorsal root ganglion. These mutants also displayed reduced inflammatory pain sensitivity on the formalin test. Further testing of pain-related phenotypes in these mutants revealed robustly increased sensitivity to systemic and spinal (but not supraspinal) morphine analgesia, and greatly increased endogenous (swim stress-induced) opioid analgesia. None of the known biological roles of Yin Yang 1 were suggestive of such a phenotype, and thus a novel player in pain modulatory systems has been identified.
Assuntos
Analgesia , Analgésicos Opioides/uso terapêutico , Morfina/uso terapêutico , Nociceptividade , Dor/genética , Fator de Transcrição YY1/genética , Animais , Células Cultivadas , Formaldeído/toxicidade , Gânglios Espinais/citologia , Camundongos , Camundongos Endogâmicos C57BL , Mutação , Canal de Sódio Disparado por Voltagem NAV1.8/genética , Canal de Sódio Disparado por Voltagem NAV1.8/metabolismo , Neurônios/metabolismo , Neurônios/fisiologia , Dor/tratamento farmacológico , Dor/etiologia , FenótipoRESUMO
Defects in expression of imprinted genes are believed to cause developmental abnormalities and play a role in carcinogenesis. To determine whether spontaneous imprinting defects may occur in mouse embryos, we studied the expression of two imprinted genes H19 and Igf2 in individual postimplantation 7.5 d.p.c. and 8.5 d.p.c. embryos. Biallelic expression of H19 was found in 1.6% of the embryos, whereas biallelic expression of Igf2 was found in 0.5% of the embryos. The loss of H19 imprinting (LOI) observed in a small fraction of early postimplantation embryos may be purely stochastic. Alternatively, since we never observed it in an inbred background, it may depend on genetic factors acting in trans. Either mechanism could explain the occurrence of polymorphic imprinting as well as the genesis of sporadic imprinting defects, including cancer. The frequency of LOI of H19 was higher than the incidence of sporadic imprinting disorders in humans (about 1 in 20,000). This contradiction may be explained by different incidence of imprinting errors in different imprinted regions of the genome, in different species, or by loss of the majority of nonmosaic embryos with imprinting defects before birth.
Assuntos
Embrião de Mamíferos/fisiologia , Impressão Genômica , Fator de Crescimento Insulin-Like II/genética , RNA não Traduzido/genética , Alelos , Animais , Deleção Cromossômica , Cruzamentos Genéticos , DNA/química , Primers do DNA/química , Desenvolvimento Embrionário , Feminino , Regulação da Expressão Gênica , Incidência , Masculino , Metilação , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos DBA , Fenótipo , Polimorfismo Genético , Gravidez , RNA Longo não Codificante , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Deviations from a Mendelian 1:1 transmission ratio have been observed in human and mouse chromosomes. With few exceptions, the underlying mechanism of the transmission-ratio distortion remains obscure. We tested a hypothesis that grandparental-origin dependent transmission-ratio distortion is related to imprinting and possibly results from the loss of embryos which carry imprinted genes with imprinting marks that have been incorrectly reset. We analyzed transmission of alleles in four regions of the human genome that carry imprinted genes presumably critical for normal embryonic growth and development: 11p15.5 (H19, IGF2, HASH2, etc.), 11p13 (WT1), 7p11-12 (GRB10), and 6q25-q27 (IGF2R), among the offspring of 31 three-generation Centre d'Etude de polymorphism Humain (CEPH) families. Deviations from expected 1:1 ratios were found in the maternal chromosomes for regions 11p15.5, 11p13, and 6q25-27 and in the paternal chromosomes for regions 11p15 and 7p11-p12. The likelihood of the results was assessed empirically to be statistically significant (p = 0.0008), suggesting that the transmission ratios in the imprinted regions significantly deviated from 1:1. We did not find deviations from a 1:1 transmission ratio in imprinted regions that are not crucial for embryo viability (13q14 and 15q11-q13). The analysis of a larger set of 51 families for the 11p15.5 region suggests that there is heterogeneity among the families with regard to the transmission of 11p15.5 alleles. The results of this study are consistent with the hypothesis that grandparental-origin dependent transmission-ratio distortion is related to imprinting and embryo loss.
Assuntos
Impressão Genômica/genética , Fatores de Transcrição , Alelos , Animais , Fatores de Transcrição Hélice-Alça-Hélice Básicos , Proteínas de Ligação a DNA/genética , Desenvolvimento Embrionário e Fetal/genética , Feminino , Proteína Adaptadora GRB10 , Humanos , Fator de Crescimento Insulin-Like II/genética , Masculino , Camundongos , Repetições de Microssatélites/genética , Mutação/genética , Linhagem , Proteínas/genética , RNA Longo não Codificante , RNA não Traduzido/genéticaRESUMO
The vitamin D binding protein/Gc-globulin (DBP) gene is a member of a multigene cluster that includes albumin (ALB), alpha-fetoprotein (AFP), and alpha-albumin/afamin (AFM). All four genes have structural and functional similarities and map to the same chromosomal regions in humans (4q11-q13), mice, and rats. An accurate physical map of the region encompassing these genes is a prerequisite for study of their respective transcriptional regulation and identification of potential shared regulatory elements. By refining the physical and meiotic maps of the 4q11-q13 region and creating a local PAC contig, the order and transcriptional orientations of these four genes were determined to be centromere-3'-DBP-5'-5'-ALB-3'-5'-AFP-3'-5'-AFM3'-telomere. The ancestral DBP gene was separated from the ALB gene by >1.5 Mb. This organization and spacing establishes a foundation for ongoing functional studies in this region.
Assuntos
Albuminas/genética , Proteínas de Transporte , Cromossomos Humanos Par 4/genética , Glicoproteínas , Meiose/genética , Família Multigênica/genética , Mapeamento Físico do Cromossomo/métodos , Proteína de Ligação a Vitamina D/genética , Animais , Marcadores Genéticos/genética , Humanos , Camundongos , Ratos , Albumina Sérica/genética , Albumina Sérica Humana , Transcrição Gênica/genética , alfa-Fetoproteínas/genéticaRESUMO
Barth syndrome (BTHS) is a rare X-linked recessive disorder characterized by cardiac and skeletal myopathy, neutropenia, and short stature. A gene for BTHS, G4.5, was recently cloned and encodes several novel proteins, named "tafazzins." Unique mutations have been found. No correlation between the location or type of mutation and the phenotype of BTHS has been found. Female carriers of BTHS seem to be healthy. This could be due to a selection against cells that have the mutant allele on the active X chromosome. We therefore analyzed X chromosome inactivation in 16 obligate carriers of BTHS, from six families, using PCR in the androgen-receptor locus. An extremely skewed X-inactivation pattern (>=95:5), not found in 148 female controls, was found in six carriers. The skewed pattern in two carriers from one family was confirmed in DNA from cultured fibroblasts. Five carriers from two families had a skewed pattern (80:20-<95:5), a pattern that was found in only 11 of 148 female controls. Of the 11 carriers with a skewed pattern, the parental origin of the inactive X chromosome was maternal in all seven cases for which this could be determined. In two families, carriers with an extremely skewed pattern and carriers with a random pattern were found. The skewed X inactivation in 11 of 16 carriers is probably the result of a selection against cells with the mutated gene on the active X chromosome. Since BTHS also shows great clinical variation within families, additional factors are likely to influence the expression of the phenotype. Such factors may also influence the selection mechanism in carriers.
Assuntos
Cardiomiopatias/genética , Triagem de Portadores Genéticos , Transtornos do Crescimento/genética , Doenças Musculares/genética , Mutação de Sentido Incorreto , Proteínas do Tecido Nervoso/genética , Neutropenia/genética , Mutação Puntual , Aberrações dos Cromossomos Sexuais , Cromossomo X , Adolescente , Adulto , Idoso , Estatura , Criança , Éxons , Feminino , Proteína do X Frágil da Deficiência Intelectual , Genes Recessivos , Humanos , Masculino , Pessoa de Meia-Idade , Linhagem , Fenótipo , Proteínas de Ligação a RNA/genética , SíndromeRESUMO
We have analyzed the transmission of maternal alleles at loci spanning the length of the X chromosome in 47 normal, genetic disease-free families. We found a significant deviation from the expected Mendelian 1:1 ratio of grandpaternal:grandmaternal alleles at loci in Xp11.4-p21.1. The distortion in inheritance ratio was found only among male offspring and was manifested as a strong bias in favor of the inheritance of the alleles of the maternal grandfather. We found no evidence for significant heterogeneity among the families, which implies that the major determinant involved in the generation of the non-Mendelian ratio is epigenetic. Our analysis of recombinant chromosomes inherited by male offspring indicates that an 11.6-cM interval on the short arm of the X chromosome, bounded by DXS538 and DXS7, contains an imprinted gene that affects the survival of male embryos.
Assuntos
Cromossomo X , Animais , Feminino , Impressão Genômica , Humanos , Masculino , Camundongos , Recombinação GenéticaRESUMO
We have analyzed X-chromosome inactivation patterns in lymphocytes of 264 females from 38 families not known to have any genetic disease. Quantitative measures of X-inactivation showed strong sister-sister correlation in the degree of departure from equal numbers of cells having each X chromosome active, suggesting heritability of this phenotype. Strong sister-sister correlation was also observed for the fraction of cells having the same parent's X chromosome active, consistent with the possibility that this trait might be controlled by a cis-acting, X-linked gene. We used a sib-pair approach to determine whether X-inactivation phenotype was linked to loci in any region of the X chromosome. Both quantitative and discrete measures of X-inactivation phenotype showed evidence of linkage to markers in the region of the X inactivation center (XIC). The quantitative measure of X-inactivation phenotype used in our study also showed linkage to loci at Xq25-q26. This study provides the first evidence for X-linked inheritance of X chromosome inactivation phenotype derived from linkage analysis in phenotypically normal human families.
Assuntos
Mapeamento Cromossômico , Mecanismo Genético de Compensação de Dose , Ligação Genética , Cromossomo X , Sequência de Bases , Primers do DNA , Feminino , Humanos , Masculino , Mães , Núcleo Familiar , Linhagem , Característica Quantitativa HerdávelRESUMO
The polar, preimplantation-embryo lethal phenotype known as the "DDK syndrome" in the mouse is the result of the complex interaction of genetic factors and a parental-origin effect. We previously observed a modest degree of transmission-ratio distortion in favor of the inheritance of DDK alleles in the Ovum mutant (Om) region of Chromosome (Chr) 11, among offspring of reciprocal F1-hybrid females and C57BL/6 males. In this study, we confirm that a significant excess of offspring inherit DDK alleles from F1 mothers and demonstrate that the preference for the inheritance of DDK alleles is not a specific bias against the C57BL/6 allele or a simple preference for offspring that are heterozygous at Om. Because none of the previous genetic models for the inheritance of the "DDK syndrome" predicted transmission-ratio distortion through F1 females, we reconsidered the possibility that the genes encoding the maternal and paternal components of this phenotype were not linked. We have examined the fertility phenotype of N2 females and demonstrate that the inter-strain fertility of these females is correlated with their genotype in the Om region. This result establishes, directly, that the genes encoding the maternal and paternal components of the DDK syndrome are genetically linked.
Assuntos
Mapeamento Cromossômico , Morte Fetal/genética , Genes Letais , Infertilidade Feminina/genética , Animais , Cruzamentos Genéticos , Feminino , Fertilidade/genética , Heterozigoto , Homozigoto , Tamanho da Ninhada de Vivíparos/genética , Masculino , Camundongos , Camundongos Endogâmicos C3H , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos , Óvulo/fisiologia , SíndromeRESUMO
One of the two X chromosomes in each somatic cell of normal human females becomes inactivated very early in embryonic development. Although the inactivation of an X chromosome in any particular somatic cell of the embryonic lineage is thought to be a stochastic and epigenetic event, a strong genetic influence on this process has been described in the mouse. We have attempted to uncover evidence for genetic control of X-chromosome inactivation in the human by examining X chromosome-inactivation patterns in 255 females from 36 three-generation pedigrees, to determine whether this quantitative character exhibits evidence of heritability. We have found one family in which all seven daughters of one male and the mother of this male have highly skewed patterns of X-chromosome inactivation, suggesting strongly that this quantitative character is controlled by one or more X-linked genes in some families.
Assuntos
RNA não Traduzido , Receptores Androgênicos/genética , Aberrações dos Cromossomos Sexuais/genética , Fatores de Transcrição/genética , Cromossomo X , Alelos , Sequência de Bases , DNA/sangue , Primers do DNA , Feminino , Impressão Genômica , Humanos , Linfócitos , Masculino , Metilação , Dados de Sequência Molecular , Linhagem , Fenótipo , Reação em Cadeia da Polimerase , Polimorfismo Genético , RNA Longo não Codificante , Valores de ReferênciaRESUMO
We determined the genotypes of > 200 offspring that are survivors of matings between female reciprocal F1 hybrids (between the DDK and C57BL/6J inbred mouse strains) and C57BL/6J males at markers linked to the Ovum mutant (Om) locus on chromosome 11. In contrast to the expectations of our previous genetic model to explain the "DDK syndrome, " the genotypes of these offspring do not reflect preferential survival of individuals that receive C57BL/6J alleles from the F1 females in the region of chromosome 11 to which the Om locus has been mapped. In fact, we observe significant transmission-ratio distortion in favor of DDK alleles in this region. These results are also in contrast to the expectations of Wakasugi's genetic model for the inheritance of Om, in which he proposed equal transmission of DDK and non-DDK alleles from F1 females. We propose that the results of these experiments may be explained by reduced expression of the maternal DDK Om allele or expression of the maternal DDK Om allele in only a portion of the ova of F1 females.
Assuntos
Mapeamento Cromossômico , Fertilidade/genética , Camundongos Endogâmicos/genética , Mutação , Animais , Cruzamentos Genéticos , Feminino , Morte Fetal/genética , Ligação Genética , Genótipo , Haplótipos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Óvulo , SíndromeRESUMO
The mammalian SR-type splicing factors ASF/SF2 and SC-35 play crucial roles in pre-mRNA splicing and have been shown to shift splice site choice in vitro. We have mapped the ASF/SF2 gene in mice and humans and the SC-35 gene in mice. Somatic cell hybrid mapping of the human ASF/SF2 gene (SFRS1 locus) reveals that it resides on chromosome 17, and fluorescence in situ hybridization refines this localization to 17q21.3-q22. Recombinant inbred mapping of the mouse ASF/SF2 gene (Sfrs1 locus) and the mouse SC-35 gene (Sfrs2 locus) demonstrates that both genes are located in a part of mouse chromosome 11 that is homologous to human chromosome 17. Mapping of Sfrs1 using F1 hybrid backcross mice between the strains C57BL/6 and DDK places Sfrs1 very near the marker D11Mit38 and indicates that the ASF/SF2 gene is closely linked to the Ovum mutant locus.
Assuntos
Mapeamento Cromossômico , Cromossomos Humanos Par 17 , Hominidae/genética , Camundongos/genética , Proteínas Nucleares/genética , Ribonucleoproteínas , Processamento Alternativo , Animais , Cricetinae/genética , Cruzamentos Genéticos , Feminino , Humanos , Células Híbridas , Hibridização in Situ Fluorescente , Masculino , Camundongos Endogâmicos/genética , Splicing de RNA , Proteínas de Ligação a RNA , Recombinação Genética , Fatores de Processamento de Serina-Arginina , Especificidade da EspécieRESUMO
We have begun a search for heritable variation in X-chromosome inactivation pattern in normal females to determine whether there is a genetic effect on the imprinting of X-chromosome inactivation in humans. We have performed a quantitative analysis of X-chromosome inactivation in lymphocytes from mothers in normal, three-generation families. Eight mothers and 12 grandmothers exhibited evidence of highly skewed patterns of X-chromosome inactivation. We observed that the male offspring of females with skewed X-inactivation patterns were three times more likely to inherit alleles at loci that were located on the inactive X chromosome (Xi) than the active X chromosome (Xa). The region of the X chromosome for which this phenomenon was observed extends from Xp11 to -Xq22. We have also examined X-chromosome inactivation patterns in 21 unaffected mothers of male bilateral sporadic retinoblastoma patients. Six of these mothers had skewed patterns of X-chromosome inactivation. In contrast to the tendency for male offspring of skewed mothers from nondisease families to inherit alleles from the inactive X chromosome, five of the six affected males inherited the androgen receptor alleles from the active X chromosome of their mother.
Assuntos
Mecanismo Genético de Compensação de Dose , Cromossomo X , Sequência de Bases , Primers do DNA , Feminino , Variação Genética , Impressão Genômica , Humanos , Masculino , Dados de Sequência Molecular , Fenótipo , Retinoblastoma/genéticaRESUMO
The incidence of free and integrated hepatitis B virus (HBV) in various human tissues is discussed. Although hepadnavirus replication is restricted to a more or less stringent host cell range, it is clear that viral integration is not restricted to any particular organ but occurs in many tissues, including the placenta, embryo, and spermatozoa. The evidence for mother-to-infant transmission of HBV is presented. Direct transfer of HBV through the germ line cells and its implication in the development of embryonal tumours are discussed. HBV may thus possess potential oncogenicity not only for those directly infected by the virus, but also for their offspring.
Assuntos
Vírus da Hepatite B/patogenicidade , DNA Viral/isolamento & purificação , Feminino , Hepatite B/etiologia , Hepatite B/transmissão , Vírus da Hepatite B/genética , Vírus da Hepatite B/fisiologia , Humanos , Recém-Nascido , Infertilidade Masculina/etiologia , Lisogenia , Masculino , Troca Materno-Fetal , Neoplasias Embrionárias de Células Germinativas/etiologia , Especificidade de Órgãos , GravidezRESUMO
Sera from children bearing embryonal tumors and from their parents were screened for the presence of hepatitis B virus (HBV) and its DNA by means of serology and molecular hybridization, respectively. Sera from tumor-bearing children and their parents both contain HBV or its DNA at average 5 times more frequently than the healthy donors or patients with non-oncological diseases. It is suggested that the presence of HBV or its DNA is caused not solely by infection during cure but also by vertical transmission from parents. The presence of HBV or its DNA might be treated as a risk factor increasing the development of embryonal tumors.
Assuntos
Hepatite B/complicações , Neoplasias Embrionárias de Células Germinativas/etiologia , Adulto , Criança , DNA de Neoplasias/análise , DNA Viral/análise , Feminino , Hepatite B/genética , Antígenos do Núcleo do Vírus da Hepatite B/análise , Antígenos de Superfície da Hepatite B/análise , Vírus da Hepatite B/genética , Vírus da Hepatite B/imunologia , Humanos , Masculino , Neoplasias Embrionárias de Células Germinativas/genética , Hibridização de Ácido NucleicoRESUMO
In spermatozoa of certain patients nucleotide sequences of hepatitis B virus were revealed in an integrated (chromosomal) and free states by means of dot- and blot-hybridization with a cloned HBV-probe. These observations support the conclusion made earlier on the lack of strict hepatotropism for HBV and, in addition, point to a possible involvement of HBV genomes in some molecular pathologies of men.
Assuntos
DNA Viral/genética , Vírus da Hepatite B/genética , Espermatozoides/análise , Sequência de Bases , DNA/análise , DNA Viral/análise , Anticorpos Anti-Hepatite B/análise , Antígenos de Superfície da Hepatite B/análise , Humanos , Masculino , Hibridização de Ácido Nucleico , Sêmen/análiseRESUMO
DNA extracted from human nonhepatic tissues (placenta and kidney) have been digested with restriction endonucleases and examined by the Southern procedure with cloned 32P-labelled DNA of hepatitis B virus (HBV). In placental DNAs of women with the history of a hepatitis B infection and in one out of four cases of patients with no known HBV exposure or manifestation, HBV-related chromosomal nucleotide sequences were detected. The integration of HBV-related sequences was observed also in human kidney DNA. Moreover, in the placenta of women who had hepatitis B infection prior to delivery, unusual unintegrated forms of HBV have been found. We conclude that HBV sequences can be found not only in hepatic tissue but also in placental and kidney DNA, both of HBV-exposed and in one case even of a nonexposed patient.
Assuntos
DNA Viral/genética , Genes Virais , Vírus da Hepatite B/genética , Hepatite B/genética , Sequência de Bases , DNA Viral/isolamento & purificação , Feminino , Hepatite B/complicações , Hepatite B/microbiologia , Vírus da Hepatite B/isolamento & purificação , Humanos , Rim/microbiologia , Fígado/microbiologia , Hibridização de Ácido Nucleico , Placenta/microbiologia , Gravidez , Complicações Infecciosas na Gravidez/genética , Complicações Infecciosas na Gravidez/microbiologiaRESUMO
We have started a cloned genomic DNA fragment approximately 7 kb long (denoted as H55) from the 7B3-4 region in the X chromosome of Drosophila melanogaster. The major part of the fragment is a single-copy sequence. It directs the synthesis of mRNA that makes up approximately 0.1% of the cytoplasmic poly(A)+ RNA from Drosophila embryos. The H55 gene is split by an intervening sequence, yielding a large single-copy exon and a small repetitive 3' exon represented by hundreds of copies in the genome. This repetitive sequence ("suffix") is also present at the 3' ends of approximately 2% of all cytoplasmic poly(A) chains.