Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 10 de 10
Filtrar
1.
Nat Genet ; 16(2): 188-90, 1997 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-9171832

RESUMEN

Genetic hearing impairment affects around 1 in every 2,000 births. The bulk (approximately 70%) of genetic deafness is non-syndromic, in which hearing impairment is not associated with any other abnormalities. Over 25 loci involved in non-syndromic deafness have been mapped and mutations in connexin 26 have been identified as a cause of non-sydromic deafness. One locus for non-syndromic recessive deafness, DFNB2 (ref. 4), has been localized to the same chromosomal region, 11q14, as one of the loci, USH1B, underlying the recessive deaf-blind syndrome. Usher syndrome type 1b, which is characterized by profound congenital sensorineural deafness, constant vestibular dysfunction and prepubertal onset of retinitis pigmentosa. Recently, it has been shown that a gene encoding an unconventional myosin, myosin VIIA, underlies the mouse recessive deafness mutation, shaker-1 (ref. 5) as well as Usher syndrome type 1b. Mice with shaker-1 demonstrate typical neuroepithelial defects manifested by hearing loss and vestibular dysfunction but no retinal pathology. Differences in retinal patterns of expression may account for the variance in phenotype between shaker-1 mice and Usher type 1 syndrome. Nevertheless, the expression of MYO7A in the neuroepithelium suggests that it should be considered a candidate for non-syndromic deafness in the human population. By screening families with non-syndromic deafness from China, we have identified two families carrying MYO7A mutations.


Asunto(s)
Sordera/genética , Genes Recesivos , Mutación , Miosinas/genética , Adulto , Animales , Cromosomas Humanos Par 11 , Dineínas , Exones , Heterocigoto , Humanos , Ratones , Persona de Mediana Edad , Miosina VIIa , Linaje , Empalme del ARN
2.
Nat Genet ; 25(4): 440-3, 2000 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-10932191

RESUMEN

As the human genome project approaches completion, the challenge for mammalian geneticists is to develop approaches for the systematic determination of mammalian gene function. Mouse mutagenesis will be a key element of studies of gene function. Phenotype-driven approaches using the chemical mutagen ethylnitrosourea (ENU) represent a potentially efficient route for the generation of large numbers of mutant mice that can be screened for novel phenotypes. The advantage of this approach is that, in assessing gene function, no a priori assumptions are made about the genes involved in any pathway. Phenotype-driven mutagenesis is thus an effective method for the identification of novel genes and pathways. We have undertaken a genome-wide, phenotype-driven screen for dominant mutations in the mouse. We generated and screened over 26,000 mice, and recovered some 500 new mouse mutants. Our work, along with the programme reported in the accompanying paper, has led to a substantial increase in the mouse mutant resource and represents a first step towards systematic studies of gene function in mammalian genetics.


Asunto(s)
Genes/fisiología , Genoma , Mutagénesis/genética , Animales , Animales Recién Nacidos , Mapeo Cromosómico , Cruzamientos Genéticos , Criopreservación , Etilnitrosourea/farmacología , Femenino , Fertilización In Vitro , Genes/efectos de los fármacos , Genes/genética , Pruebas Hematológicas , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C3H , Actividad Motora/genética , Mutagénesis/efectos de los fármacos , Mutágenos/farmacología , Mutación , Fenotipo , Factores de Tiempo , Destete
3.
Biochim Biophys Acta ; 1172(1-2): 5-11, 1993 Feb 20.
Artículo en Inglés | MEDLINE | ID: mdl-8439571

RESUMEN

DNA fragments including the promoter region of the major ribosomal RNA gene of Trypanosoma brucei (r-promoter) were identified and subcloned using a synthetic oligonucleotide probe corresponding to the putative core promoter. These fragments were used in mobility shift assays with proteins extracted from T. brucei nuclei, and demonstrated the presence in 0.15 M NaCl extracts of protein(s) with specific binding affinities for the r-promoter region. Binding was stable in the presence of a 100-fold excess of competitor DNA, and occurred at the relatively low salt concentrations (< 50 mM NaCl) characteristic of many enzyme activity optima in this organism. A control DNA fragment not including the r-promoter region was not retarded in the mobility shift assays, and the r-promoter-binding activity had a molecular weight of about 140,000. Nuclear extracts from T. brucei contained large amounts of DNase activity, and the promoter-binding proteins were partially purified from the crude extract using ammonium sulphate precipitation, sephacryl S-200 and Heparin-sepharose chromatography.


Asunto(s)
ADN Protozoario/genética , ADN Ribosómico/genética , Proteínas de Unión al ADN/metabolismo , Regiones Promotoras Genéticas , ARN Ribosómico/genética , Trypanosoma brucei brucei/genética , Animales , Secuencia de Bases , Southern Blotting , Núcleo Celular/metabolismo , Cromatografía de Afinidad , Cromatografía en Gel , Clonación Molecular , ADN Protozoario/aislamiento & purificación , ADN Ribosómico/aislamiento & purificación , Proteínas de Unión al ADN/aislamiento & purificación , Datos de Secuencia Molecular , Peso Molecular , Oligodesoxirribonucleótidos , Mapeo Restrictivo , Trypanosoma brucei brucei/metabolismo
4.
Ann Otol Rhinol Laryngol Suppl ; 168: 59-62, 1997 May.
Artículo en Inglés | MEDLINE | ID: mdl-9153119

RESUMEN

Hearing-impaired mouse mutants not only are good models for human hereditary deafness, but also are extremely useful for understanding the molecular basis of the cochlear defect. We describe here how we identified the gene responsible for the deafness and vestibular defects in the shaker-1 mouse mutant as a myosin VII gene. Three different mutations, all causing the same phenotype in different lines of mouse, were found, providing good evidence that we had, indeed, found the correct gene. The same gene was subsequently found to be involved in Usher's syndrome type 1B, which features deafness, vestibular dysfunction, and progressive retinitis pigmentosa. The myosin VII gene is expressed in sensory hair cells, but not in supporting cells or neurons. We are investigating the role of myosin VII in hair cell development and function. Analysis of the different mutant stocks suggests it has at least two functions. First it is involved in the development and maintenance of the stereocilia bundle. Second, it has a role in inner hair cell function. No evidence of retinal degeneration like that in Usher's syndrome has been found in the shaker-1 mutants so far studied. The benefits of understanding the function of the gene for families with Usher's type 1B are discussed. This gene is the first to be identified as causing the most common type of disorder in human hearing impairment, neuroepithelial abnormalities, and suggests a new class of candidate genes for involvement in such defects.


Asunto(s)
Sordera/genética , Animales , Aberraciones Cromosómicas , Trastornos de los Cromosomas , Clonación Molecular , Sordera/fisiopatología , Células Ciliadas Auditivas/fisiopatología , Ratones , Ratones Mutantes , Miosinas/genética , Órgano Espiral/fisiopatología
5.
Br J Audiol ; 33(5): 279-83, 1999 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-10890141

RESUMEN

The availability of mouse mutant models for known human deafness loci is limited. Moreover, it is unlikely that the current mouse archives hold mutants for the full panoply of genes involved in auditory system development and transduction. A large-scale ENU mutagenesis is currently underway to increase significantly the number of mouse deafness mutants available, employing specific screens for both deafness and balance defects. In the MRC Harwell screen, 13 mice have been identified so far with deafness, a balance defect or both. Mutagenized mice from the programme are also being used to search for modifiers of a known deafness gene, myosin VIIA (mutated in the Shaker 1 mutant mouse). The progress and encouraging results of the programme indicate that the combination of ENU mutagenesis and effective phenotype screens will lead to a significant contribution to the understanding of the genes and mechanisms involved in hereditary deafness.


Asunto(s)
Sordera/genética , Etilnitrosourea/farmacología , Mutagénesis/efectos de los fármacos , Mutagénesis/genética , Animales , Modelos Animales de Enfermedad , Ratones , Mutación Puntual/efectos de los fármacos , Mutación Puntual/genética
6.
Mamm Genome ; 10(5): 513-9, 1999 May.
Artículo en Inglés | MEDLINE | ID: mdl-10337627

RESUMEN

The whirler (wi) mutation on mouse Chromosome (Chr) 4 results in an autosomal recessive neuroepithelial deafness and vestibular dysfunction exhibited as a characteristic shaker-waltzer behavior (deafness, circling, and head-bobbing). We have constructed a genetic linkage map across the wi region in both an interspecific [(wi/wi x CAST/Ei)F1 x wi/wi] backcross (n = 817) and an intraspecific [(wi/wi x CBA/Ca)F1 x wi/wi)] backcross (n = 335). In the interspecific backcross, wi was found to be non-recombinant with Orm1, 0.12 cM distal of D4Mit87 and Ambp, and 0.12 cM proximal of CD301. In the intraspecific backcross, wi was found to be non-recombinant with Orm1 and D4Mit244, 0.3 cM distal of Mup1, and 0.6 cM proximal of Tnc. We also report a family from the interspecific backcross that shows evidence of multiple recombinations across the region of mouse Chr 4 around the wi locus. These rearrangements appear specific to both the region and the family.


Asunto(s)
Mapeo Cromosómico , Mutación , Animales , Cruzamientos Genéticos , Ratones , Ratones Endogámicos CBA , Ratones Mutantes , Especificidad de la Especie
7.
Nature ; 374(6517): 60-1, 1995 Mar 02.
Artículo en Inglés | MEDLINE | ID: mdl-7870171

RESUMEN

Usher syndrome represents the association of a hearing impairment with retinitis pigmentosa and is the most frequent cause of deaf-blindness in humans. It is inherited as an autosomal recessive trait which is clinically and genetically heterogeneous. Some patients show abnormal organization of microtubules in the axoneme of their photoreceptors cells (connecting cilium), nasal ciliar cells and sperm cells, as well as widespread degeneration of the organ of Corti. Usher syndrome type 1 (USH1) is characterized by a profound congenital sensorineural hearing loss, constant vestibular dysfunction and prepubertal onset of retinitis pigmentosa. Of three different genes responsible for USH1. USH1B maps to 11q13.5 (ref. 10) and accounts for about 75% of USH1 patients. The mouse deafness shaker-1 (sh1) mutation has been localized to the homologous murine region. Taking into account the cytoskeletal abnormalities in USH patients, the identification of a gene encoding an unconventional myosin as a candidate for shaker-1 (ref. 14) led us to consider the human homologue as a good candidate for the gene that is defective in USH1B. Here we present evidence that a gene encoding myosin VIIA is responsible for USH1B. Two different premature stop codons, a six-base-pair deletion and two different missense mutations were detected in five unrelated families. In one of these families, the mutations were identified in both alleles. These mutations, which are located at the amino-terminal end of the motor domain of the protein, are likely to result in the absence of a functional protein. Thus USH1B appears as a primary cytoskeletal protein defect. These results implicate the genes encoding other unconventional myosins and their interacting proteins as candidates for other genetic forms of Usher syndrome.


Asunto(s)
Sordera/genética , Mutación , Miosinas/genética , Retinitis Pigmentosa/genética , Adulto , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Niño , Cromosomas Humanos Par 11 , ADN , Análisis Mutacional de ADN , Sordera/congénito , Femenino , Humanos , Masculino , Ratones , Datos de Secuencia Molecular , Homología de Secuencia de Aminoácido , Porcinos , Síndrome
8.
Nature ; 374(6517): 62-4, 1995 Mar 02.
Artículo en Inglés | MEDLINE | ID: mdl-7870172

RESUMEN

Genetic deafness is common, affecting about 1 in 2,000 births. Many of these show primary abnormalities of the sensory neuroepithelia of the inner ear, as do several hearing-impaired mouse mutants, suggesting that genes involved in sensory transduction could be affected. Here we report the identification of one such gene, the mouse shaker-1 (sh1) gene. Shaker-1 homozygotes show hyperactivity, head-tossing and circling due to vestibular dysfunction, together with typical neuroepithelial-type cochlear defects involving dysfunction and progressive degeneration of the organ of Corti. The sh1 gene encodes an unconventional myosin molecule of the type VII family. Three mutations are described, two mis-sense mutations and a splice acceptor site mutation, all in the region encoding the myosin head. The myosin type VII molecule encoded by sh1 is the first molecule to be identified that is known, by virtue of its mutations, to be involved in auditory transduction.


Asunto(s)
Sordera/genética , Miosinas/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Cromosomas Artificiales de Levadura , Cartilla de ADN , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos CBA , Datos de Secuencia Molecular , Mutación , Proteínas del Tejido Nervioso/genética , Proteína Marcadora Olfativa
9.
Genes Funct ; 1(3): 191-203, 1997 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-9680294

RESUMEN

The shaker-1 (Myo7a) mouse deafness locus is encoded by an unconventional myosin gene: myosin VIIA [Gibson, Walsh, Mburu, Varela, Brown, Antonio, Biesel, Steel and Brown (1995) Nature (London) 374, 62-64]. The myosin VIIA gene is expressed in hair cells in the cochlea, where it is thought to function in the development of the critical neuroepithelium where auditory transduction takes place. In order to understand better the function of myosin VIIA, we have determined the complete sequence of the mouse myosin VIIA cDNA and employed the wild-type sequence for mutational analysis of a number of shaker-1 alleles. Analysis of the mouse myosin VIIA tail sequence demonstrates a large internal repeat with regions of similarity to myosins IV, X and XII as well as members of the band 4.1 family. In addition, the myosin VIIA repeats are similar along their entire length to a tail domain from a plant kinesin. The mouse myosin VIIA tail also contains a putative Src homology 3 (SH3) domain. Along with three previously reported shaker-1 mutations, mutations for seven shaker-1 alleles in total have now been identified. The mutational changes have been analysed in terms of their predicted effect on both myosin motor head and tail domain function and the predictions related to the known phenotypes of the shaker-1 alleles. Five of the mutations lie in the motor head, and analysis of their likely effect on myosin head structure correlates well with the known severity of the shaker-1 alleles. Of the two mutations in the tail, one is a missense mutation within the kinesin and myosin IV, X and XII homology domains that substitutes a conserved amino acid and leads to a severe deafness phenotype. This and other data suggest that myosin VIIA may have properties of a myosin-motor-kinesin-tail hybrid and be involved in membrane turnover within the actin-rich environment of the apical hair cell surface.


Asunto(s)
Análisis Mutacional de ADN , Sordera/genética , Miosinas/genética , Secuencia de Aminoácidos , Animales , Clonación Molecular , ADN Complementario/genética , Dineínas , Genes/genética , Ratones , Ratones Mutantes Neurológicos , Modelos Moleculares , Datos de Secuencia Molecular , Miosina VIIa , Miosinas/química , ARN Mensajero/genética , Secuencias Repetitivas de Ácidos Nucleicos/genética , Alineación de Secuencia , Homología de Secuencia de Ácido Nucleico , Transcripción Genética/genética , Dominios Homologos src
10.
Hum Mol Genet ; 10(5): 507-12, 2001 Mar 01.
Artículo en Inglés | MEDLINE | ID: mdl-11181574

RESUMEN

The Notch signalling pathway has recently been implicated in the development and patterning of the sensory epithelium in the cochlea, the organ of Corti. As part of an ongoing large-scale mutagenesis programme to identify new deaf or vestibular mouse mutants, we have identified a novel mouse mutant, slalom, which shows abnormalities in the patterning of hair cells in the organ of Corti and missing ampullae, structures that house the sensory epithelia of the semicircular canals. We show that the slalom mutant carries a mutation in the Jagged1 gene, implicating a new ligand in the signalling processes that pattern the inner ear neuro-epithelium.


Asunto(s)
Tipificación del Cuerpo , Proteínas de la Membrana/genética , Órgano Espiral/embriología , Animales , Secuencia de Bases , Proteínas de Unión al Calcio , Clonación Molecular , Cartilla de ADN , Homocigoto , Péptidos y Proteínas de Señalización Intercelular , Proteína Jagged-1 , Ratones , Ratones Endogámicos C3H , Microscopía Electrónica de Rastreo , Mutación , Defectos del Tubo Neural/genética , Proteínas Serrate-Jagged
SELECCIÓN DE REFERENCIAS
DETALLE DE LA BÚSQUEDA