RESUMEN
To illuminate the function and evolutionary history of both genomes, we sequenced mouse DNA related to human chromosome 19. Comparative sequence alignments yielded confirmatory evidence for hypothetical genes and identified exons, regulatory elements, and candidate genes that were missed by other predictive methods. Chromosome-wide comparisons revealed a difference between single-copy HSA19 genes, which are overwhelmingly conserved in mouse, and genes residing in tandem familial clusters, which differ extensively in number, coding capacity, and organization between the two species. Finally, we sequenced breakpoints of all 15 evolutionary rearrangements, providing a view of the forces that drive chromosome evolution in mammals.
Asunto(s)
Cromosomas Humanos Par 19/genética , Secuencia Conservada/genética , Evolución Molecular , Animales , Rotura Cromosómica/genética , Mapeo Contig , ADN Satélite/genética , Exones/genética , Etiquetas de Secuencia Expresada , Dosificación de Gen , Orden Génico/genética , Ligamiento Genético/genética , Genoma , Humanos , Elementos de Nucleótido Esparcido Largo/genética , Ratones , Familia de Multigenes/genética , Sistemas de Lectura Abierta/genética , Filogenia , Alineación de Secuencia , Análisis de Secuencia de ADN , Elementos de Nucleótido Esparcido Corto/genética , Secuencias Repetidas Terminales/genéticaRESUMEN
The dark-adapted electroretinogram (ERG) of patients with Duchenne and Becker muscular dystrophy (DMD/BMD) shows a marked reduction in b-wave amplitude. Genotype-phenotype studies of mouse models for DMD show position-specific effects of the mutations upon the phenotype: mice with 5' defects of dystrophin have normal ERGs, those with defects in the central region have a normal b-wave amplitude associated with prolonged implicit times for both the b-wave and oscillatory potentials, and mice with 3' defects have a phenotype similar to that seen in DMD/BMD patients. The mouse studies suggest a key role for the carboxyl terminal dystrophin isoform, Dp260, in retinal electrophysiology. We have undertaken a systematic evaluation of DMD/BMD patients through clinical examination and review of the literature in order to determine whether the position-specific effects of mutations noted in the mouse are present in man. We have found that, in man, a wider variation of DMD defects correlate with reductions in the b-wave amplitude. Individuals with normal ERGs have mutations predominantly located 5' of the transcript initiation site of Dp260. Our results suggest that the most important determinant in the ERG b-wave phenotype is the mutation position, rather than muscle disease severity. Forty-six per cent of patients with mutations 5' of the Dp260 transcript start site have abnormal ERGs, as opposed to 94% with more distal mutations. The human genotype-phenotype correlations are consistent with a role for Dp260 in normal retinal electrophysiology and may also reflect the expression of other C-terminal dystrophin isoforms and their contributions to retinal signal transmission.
Asunto(s)
Distrofina/genética , Electrorretinografía , Distrofias Musculares/genética , Mutación , Exones , Eliminación de Gen , Genotipo , Humanos , Modelos Genéticos , Distrofias Musculares/fisiopatología , Fenotipo , Reacción en Cadena de la Polimerasa , Regiones Promotoras Genéticas , ARN Mensajero/análisisRESUMEN
OBJECTIVES/HYPOTHESIS: Sensorineural hearing loss has been found in association with inherited muscular dystrophies in humans and in mouse models. An increased brainstem auditory evoked response threshold has been previously reported in the dystrophin-deficient mdx mouse model for Duchenne muscular dystrophy, suggesting that full-length dystrophin (Dp427) is involved in hearing. The objective of the present study was to confirm cochlear dysfunction with this gene defect and determine whether the shorter carboxyl terminus isoforms of dystrophin are also critical in maintaining normal hearing. STUDY DESIGN: Case controlled. Animal model. METHODS: Auditory brainstem response (ABR) audiometry to pure tones was used to evaluate cochlear function. Fourteen mdx, 4 mdx(Cv3), and 13 age-matched control (C57BL/6J and C57BL/10ScSn) male mice were tested at 5 weeks and 11 weeks of age. The ABR thresholds to tone-burst stimuli at 4, 8, 16, and 32 kHz were obtained for each ear and statistically compared (ANOVA) for potential group differences. RESULTS: Both mdx and mdx(Cv3) mice demonstrated normal ABR thresholds when compared with controls. CONCLUSIONS: Both mdx and mdx(Cv3) mouse models have normal hearing by ABR. The authors' data suggest that dystrophin and its carboxyl terminus isoforms do not play a critical role in hearing in the mouse. This was unexpected, as previous studies using the brainstem auditory evoked response method suggested that the mdx mouse has an increased threshold for hearing.
Asunto(s)
Cóclea/fisiología , Modelos Animales de Enfermedad , Distrofina/deficiencia , Distrofia Muscular Animal/metabolismo , Distrofia Muscular Animal/fisiopatología , Animales , Umbral Auditivo , Estudios de Casos y Controles , Distrofina/genética , Potenciales Evocados Auditivos , Potenciales Evocados Auditivos del Tronco Encefálico , Genotipo , Audición/fisiología , Pérdida Auditiva Sensorineural/diagnóstico , Masculino , Ratones , Ratones Endogámicos mdx , Distrofia Muscular Animal/genética , Isoformas de ProteínasRESUMEN
Duchenne and Becker muscular dystrophy patients have mutations in the dystrophin gene. Most show reduced b-wave amplitudes in the dark-adapted electroretinogram (ERG). We studied normal C57BL/6J mice and five X-linked muscular dystrophy strains with different dystrophin mutations to determine whether the location of the mutation within the gene affects the mouse ERG and to correlate such effects with dystrophin isoform expression. Amplitudes and implicit times were measured for a-waves, b-waves, and digitally filtered oscillatory potentials. mdx and mdxCv5 mice, with mutations near the amino terminus and lacking expression of Dp427, had ERGs similar to those of C57BL/6J mice. mdxCv2 and mdxCv4 mice, with mutations in the center of dystrophin and who do not express isoforms Dp427, Dp260, or Dp140 (mdxCv4), had increased b-wave and oscillatory potential implicit times. mdxCv3 mice, with a mutation near the carboxy terminus resulting in deficiency of all dystrophin isoforms, had increased b-wave and oscillatory potential implicit times and reduced scotopic b-wave amplitudes. Fitting the a-wave data to a transduction activation phase mathematical model showed normal responses for all phenotypes, suggesting that the b-wave delays are due to defects beyond the rod outer segment, most likely at the rod to on-bipolar cell synapse. The variation in the ERG phenotype with the position of the dystrophin gene mutation suggests that there are different contributions by each isoform to retinal electrophysiology. Although Dp427 and Dp140 isoforms do not appear to be important contributors to the ERG, lack of Dp260 and possibly Dp71 isoforms is associated with an abnormal ERG.
Asunto(s)
Distrofina/genética , Distrofina/metabolismo , Distrofia Muscular Animal/genética , Distrofia Muscular Animal/fisiopatología , Neuronas/fisiología , Retina/fisiopatología , Transducción de Señal/fisiología , Animales , Electrorretinografía , Potenciales Evocados , Genotipo , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos mdx , Distrofia Muscular Animal/patología , Neuronas/citología , Neuronas/patología , Fenotipo , Estimulación Luminosa , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Mapeo Restrictivo , Retina/patología , Retina/fisiología , Cromosoma XRESUMEN
Duchenne muscular dystrophy (DMD) is caused by a defect in a 427-kDa membrane-associated protein: dystrophin. The DMD gene also encodes several shorter isoforms which are believed to participate in nonmuscle manifestations of DMD, including abnormal retinal electrophysiology, dilated cardiomyopathy, mental retardation, and hearing defects. The purpose of this work was to determine the normal tissue expression of full-length dystrophin (Dp427) and the dystrophin isoforms Dp260, Dp140, Dp116, and Dp71, to aid in understanding what roles these isoforms might play in DMD nonmuscle manifestations. RT-PCR was performed on mRNA isolated from wild-type C57BL/6J mouse tissues, including brain, cardiac muscle, eye, intestine, kidney, liver, lung, skeletal muscle, spleen, stomach, testis, thymus, and uterus. RT-PCR amplification demonstrated that the isoforms were in a number of tissues which had not been revealed by previous Western and Northern blot analyses. Dp427 was expressed at equal levels in all tissues. Dp260 and Dp140 were present in all tissues tested, but the levels of expression varied. Dp116 was expressed in a subset of tissues and levels of expression varied. Dp71 was constitutively expressed in all tissues, suggesting that this isoform plays a basic role in normal tissue function. The expanded tissue distribution supports the hypothesis that dystrophin isoforms serve essential and unique functions, necessitating further investigation into their potential roles in DMD nonmuscle manifestations.
Asunto(s)
Distrofina/genética , Distrofina/metabolismo , Distrofias Musculares/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Actinas/genética , Animales , Cartilla de ADN , Electroforesis en Gel de Agar , Femenino , Isomerismo , Masculino , Ratones , Ratones Endogámicos C57BL , Distribución TisularRESUMEN
Transcribing sign communication used simultaneously with spoken English presents investigators with a unique problem: the singular quality of the bimodal communicative interactions cannot be accurately depicted using accepted conventions for recording either spoken or signed language samples. This article proposes guidelines for transcribing such data. The need for guidelines arose during an earlier study of the language development of a hearing child of deaf parents. To meet the immediate needs of that study, rules and conventions from previous studies were combined with newly generated ones, resulting in the guidelines proposed in this article. The guidelines can be applied to data in which intermodal linguistic influence is suspected.
Asunto(s)
Lengua de Signos , Habla , Escritura , Métodos de Comunicación Total , Humanos , MétodosRESUMEN
In five experiments, 10-month-olds were habituated to exemplars of a form category and tested for categorization in paired-comparison trials involving in-category versus out-of-category stimuli. Across these experiments, color was systematically manipulated during habituation and/or test trials. Infants categorized form when color was either held constant or varied during habituation, but failed to categorize form when exposed to color-constant stimuli during habituation and tested for categorization with novel-color form exemplars. Two subsequent experiments traced this failure to the narrow experience of exposure to color-constant exemplars during habituation. These results suggest that (a) infants' internal representation for a category will not include a stimulus dimension not varied in the exemplars from which the category was derived, but (b) if variation in that dimension is experienced, exemplars constructed of novel instances of that dimension will still be regarded as belonging to the category.