RESUMO
LCA is a severe retinal dystrophy characterised by an onset of symptoms before the age of 6 months, visual acuity below 201/400, searching nystagmus, sluggish pupillary reactions and no detectable responses on electrography. The visual fields are usually not measurable. LCA is genetically heterogeneous and is usually inherited in an autosomal recessive fashion. Seven genes have been reported to be mutated in LCA patients (AIPL1, CRB1, CRX, GUCY2D, RDH12, RPE65 and RPGRIP1). Each gene is responsible for a fraction of LCA patients. Mutations in these seven genes are estimated to underlie approximately 40-50% of LCA cases. Molecular genetic research is crucial to unravel the remaining genetic causes of this disabling disease.
Assuntos
Genes Recessivos , Mutação , Atrofia Óptica Hereditária de Leber/genética , Cegueira/genética , Cegueira/patologia , Eletrorretinografia , Humanos , Atrofia Óptica Hereditária de Leber/patologia , Degeneração Retiniana/genética , Degeneração Retiniana/patologia , Acuidade VisualRESUMO
Mutations in the transmembrane channel-like gene 1 (TMC1) cause prelingual autosomal recessive (DFNB7/11) and postlingual progressive autosomal dominant (DFNA36) nonsyndromic hearing loss. To determine the genetic causes of autosomal recessive nonsyndromic hearing loss (ARNSHL) in the northeast and east of Turkey, 65 unrelated families without mutations in the protein coding region of the GJB2 (GJB2-negative) were analyzed. A genomewide scan for homozygosity and linkage analysis in one of these families revealed a 13.2 cM critical region between D9S273 and D9S153 at chromosome 9p13.2-q21.31 with a maximum two-point lod score of 4.00 at theta=0.0 for marker D9S175. TMC1 is in this critical region. Homozygosity screening with intragenic markers for TMC1 in the remaining 64 families suggested involvement of this gene in three additional families. Subsequent sequencing of TMC1 in these four families revealed four novel homozygous mutations, c.776A>G [p.Tyr259Cys], c.821C>T [p.Pro274Leu], c.1334G>A [p.Arg445His], and c.1083_1087delCAGAT [p.Arg362ProfrX6]. Our results indicate that TMC1 mutations account for at least 6% (4/65) of ARNSHL in GJB2-negative Turkish families from the northeast and east of Turkey.
Assuntos
Mutação da Fase de Leitura , Perda Auditiva/genética , Proteínas de Membrana/genética , Mutação de Sentido Incorreto , Sequência de Aminoácidos , Conexina 26 , Conexinas/genética , Análise Mutacional de DNA , Feminino , Ligação Genética , Testes Genéticos , Haplótipos , Perda Auditiva/congênito , Humanos , Masculino , Proteínas de Membrana/química , Dados de Sequência Molecular , Linhagem , Alinhamento de Sequência , TurquiaRESUMO
Homozygosity mapping and linkage analysis in a Turkish family with autosomal recessive prelingual sensorineural hearing loss revealed a 15-cM critical region at 17q25.1-25.3 flanked by the polymorphic markers D17S1807 and D17S1806. The maximum two-point lod score was 4.07 at theta=0.0 for the marker D17S801. The linkage interval contains the Usher syndrome 1G gene (USH1G) that is mutated in patients with Usher syndrome (USH) type 1g and encodes the SANS protein. Mutation analysis of USH1G led to the identification of a homozygous missense mutation D458V at the -3 position of the PDZ binding motif of SANS. This mutation was also present homozygously in one out of 64 additional families from Turkey with autosomal recessive nonsyndromic hearing loss and heterozygously in one out of 498 control chromosomes. By molecular modeling, we provide evidence that this mutation impairs the interaction of SANS with harmonin. Ophthalmologic examination and vestibular evaluation of patients from both families revealed mild retinitis pigmentosa and normal vestibular function. These results suggest that these patients suffer from atypical USH.
Assuntos
Perda Auditiva Neurossensorial/genética , Mutação de Sentido Incorreto , Proteínas do Tecido Nervoso/genética , Síndromes de Usher/genética , Motivos de Aminoácidos , Sequência de Aminoácidos , Audiometria de Tons Puros , Mapeamento Cromossômico , Cromossomos Humanos Par 17 , Consanguinidade , Análise Mutacional de DNA , Éxons , Feminino , Genes Recessivos , Ligação Genética , Marcadores Genéticos , Haplótipos , Homozigoto , Humanos , Ligação de Hidrogênio , Escore Lod , Masculino , Modelos Moleculares , Dados de Sequência Molecular , Proteínas do Tecido Nervoso/química , Linhagem , Polimorfismo Genético , Estrutura Terciária de Proteína , Sequências de Repetição em Tandem , Turquia/epidemiologiaRESUMO
Mutations in the ABCA4 gene cause Stargardt disease (STGD), most cases with autosomal recessive (ar) cone-rod dystrophy (CRD), and some cases with atypical ar retinitis pigmentosa (arRP). We found compound heterozygous ABCA4 mutations in two unrelated patients with STGD and homozygous splice site mutations in their 2nd and 4th degree cousins with RP. Some ABCA4 mutations display strong founder effects. In Dutch and German STGD patients, the 768G > T mutation is present in 8% and 0.6% of ABCA4 alleles respectively. Vice versa, the complex L541P;A1038V allele is found in 70% of ABCA4 alleles in German STGD patients but absent in Dutch patients. As approximately 70% of ABCA4 mutations are known, a microarray-based analysis of known ABCA4 gene variants allows routine DNA diagnostics in Caucasian patients. Mutations in the CRB1 gene underlie RP12, some cases with classic arRP, 55% of cases with RP and Coats-like exudative vasculopathy, and 13% of patients with Leber congenital amaurosis (LCA), rendering CRB1 a significant cause of autosomal recessive retinal dystrophy. Different combinations of mutations in ABCA4 or CRB1 can be correlated with disease severity, suggesting that small increments of protein activities in patients might have significant therapeutic effects. Mouse and Drosophila studies strongly suggest that both patient groups might benefit from reduced light exposure and therefore should be detected as early as possible using molecular techniques.
Assuntos
Transportadores de Cassetes de Ligação de ATP/metabolismo , Cegueira/genética , Proteínas do Olho , Proteínas de Membrana/metabolismo , Proteínas do Tecido Nervoso , Transportadores de Cassetes de Ligação de ATP/genética , Transportadores de Cassetes de Ligação de ATP/isolamento & purificação , Cegueira/metabolismo , Genótipo , Humanos , Proteínas de Membrana/genética , Mutação , Análise de Sequência com Séries de Oligonucleotídeos , FenótipoRESUMO
Linkage analysis in a multigenerational family with autosomal dominant hearing loss yielded a chromosomal localisation of the underlying genetic defect in the DFNA20/26 locus at 17q25-qter. The 6-cM critical region harboured the gamma-1-actin (ACTG1) gene, which was considered an attractive candidate gene because actins are important structural elements of the inner ear hair cells. In this study, a Thr278Ile mutation was identified in helix 9 of the modelled protein structure. The alteration of residue Thr278 is predicted to have a small but significant effect on the gamma 1 actin structure owing to its close proximity to a methionine residue at position 313 in helix 11. Met313 has no space in the structure to move away. Moreover, the Thr278 residue is highly conserved throughout eukaryotic evolution. Using a known actin structure the mutation could be predicted to impair actin polymerisation. These findings strongly suggest that the Thr278Ile mutation in ACTG1 represents the first disease causing germline mutation in a cytoplasmic actin isoform.
Assuntos
Actinas/genética , Predisposição Genética para Doença , Perda Auditiva Neurossensorial/genética , Mutação de Sentido Incorreto , Actinas/química , Sequência de Bases , Feminino , Humanos , Masculino , Modelos Moleculares , Dados de Sequência Molecular , Linhagem , Análise de SequênciaRESUMO
Genetic variation in the ABCR (ABCA4) gene has been associated with five distinct retinal phenotypes, including Stargardt disease/fundus flavimaculatus (STGD/FFM), cone-rod dystrophy (CRD), and age-related macular degeneration (AMD). Comparative genetic analyses of ABCR variation and diagnostics have been complicated by substantial allelic heterogeneity and by differences in screening methods. To overcome these limitations, we designed a genotyping microarray (gene chip) for ABCR that includes all approximately 400 disease-associated and other variants currently described, enabling simultaneous detection of all known ABCR variants. The ABCR genotyping microarray (the ABCR400 chip) was constructed by the arrayed primer extension (APEX) technology. Each sequence change in ABCR was included on the chip by synthesis and application of sequence-specific oligonucleotides. We validated the chip by screening 136 confirmed STGD patients and 96 healthy controls, each of whom we had analyzed previously by single strand conformation polymorphism (SSCP) technology and/or heteroduplex analysis. The microarray was >98% effective in determining the existing genetic variation and was comparable to direct sequencing in that it yielded many sequence changes undetected by SSCP. In STGD patient cohorts, the efficiency of the array to detect disease-associated alleles was between 54% and 78%, depending on the ethnic composition and degree of clinical and molecular characterization of a cohort. In addition, chip analysis suggested a high carrier frequency (up to 1:10) of ABCR variants in the general population. The ABCR genotyping microarray is a robust, cost-effective, and comprehensive screening tool for variation in one gene in which mutations are responsible for a substantial fraction of retinal disease. The ABCR chip is a prototype for the next generation of screening and diagnostic tools in ophthalmic genetics, bridging clinical and scientific research.
Assuntos
Transportadores de Cassetes de Ligação de ATP/genética , Análise Mutacional de DNA/métodos , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Doenças Retinianas/genética , Variação Genética , Genótipo , Humanos , Polimorfismo Genético , Reprodutibilidade dos TestesRESUMO
To identify novel genes that are expressed specifically or preferentially in the cochlea, we constructed a cDNA library enriched for human cochlear cDNAs using a suppression subtractive hybridization technique. We analyzed 2640 clones by sequencing and BLAST similarity searches. One hundred and fifty-five different cDNA fragments mapped in nonsyndromic hearing impairment loci for which the causative gene has not been cloned yet. Approximately 30% of the clones show no similarity to any known human gene or expressed sequence tag (EST). Clones mapping in nonsyndromic deafness loci and a selection of clones that represent novel ESTs were analyzed by reverse transcriptase-polymerase chain reaction (RT-PCR) of RNA derived from 12 human fetal tissues. Our data suggest that a quarter of the novel genes in our library are preferentially expressed in fetal cochlea. These may play a physiologically important role in the hearing process and represent candidate genes for hereditary hearing impairment.
Assuntos
Cóclea/metabolismo , DNA Complementar/genética , Expressão Gênica , Sequência de Bases , Mapeamento Cromossômico , Clonagem Molecular , DNA Complementar/química , DNA Complementar/metabolismo , Surdez/genética , Perfilação da Expressão Gênica , Biblioteca Gênica , Humanos , Hibridização In Situ/métodos , Dados de Sequência Molecular , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNAAssuntos
Efeito Fundador , Proteínas/genética , Degeneração Retiniana/genética , Substituição de Aminoácidos , Proteínas de Transporte , DNA/química , DNA/genética , Análise Mutacional de DNA , Progressão da Doença , Proteínas do Olho , Saúde da Família , Feminino , Variação Genética , Heterozigoto , Histidina/genética , Homozigoto , Humanos , Masculino , Mutação , Países Baixos , Linhagem , Degeneração Retiniana/patologia , Tirosina/genética , cis-trans-IsomerasesAssuntos
Surdez/genética , Doenças Genéticas Ligadas ao Cromossomo X/genética , Perda Auditiva Condutiva/genética , Perda Auditiva Neurossensorial/genética , Estribo/anormalidades , Audiometria , Limiar Auditivo , Condução Óssea , Surdez/congênito , Surdez/diagnóstico , Surdez/patologia , Orelha Interna/patologia , Perda Auditiva Condutiva/diagnóstico , Perda Auditiva Neurossensorial/diagnóstico , Humanos , Linhagem , Cirurgia do Estribo/efeitos adversosRESUMO
Autosomal recessive Stargardt disease is caused by mutations in the ABCA4 gene. Mutations in ABCA4 are also found in two-thirds of cases with autosomal recessive cone-rod dystrophy, and a small fraction of patients with autosomal recessive retinitis pigmentosa. Patients with autosomal recessive retinitis pigmentosa, the most severe of these three phenotypes, invariably carry ABCA4 inactivating mutations; patients with autosomal recessive cone-rod dystrophy and Stargardt disease carry combinations of mutations that do not completely inactivate the retina specific 'ATP-binding cassette transporter' (ABCR) protein. DNA diagnostics is complicated by the high allelic heterogeneity and the uncertainty as to whether some ABCA4 variants are pathological. Nevertheless, ABCA4 mutation analysis is particularly important for patients with cone-rod dystrophy to confirm the autosomal recessive mode of inheritance.
Assuntos
Transportadores de Cassetes de Ligação de ATP/genética , Degeneração Macular/genética , Mutação , Retinose Pigmentar/genética , Fundo de Olho , Genes Recessivos/genética , Humanos , Países Baixos , Degeneração Retiniana/genéticaRESUMO
A Dutch family with autosomal dominant retinitis pigmentosa (adRP) displayed a phenotype characterized by an early age of onset, a diffuse loss of rod and cone sensitivity, and constricted visual fields (type I). One male showed a mild progression of the disease. Linkage analysis showed cosegregation of the genetic defect with markers from chromosome 17p13.1-p13.3, a region overlapping the RP13 locus. The critical interval of the RP locus as defined in this family was flanked by D17S926 and D17S786, with a maximal lod score of 4.2 (theta = 0.00) for marker D17S1529. Soon after the mapping of the underlying defect to the 17p13 region, a missense mutation (6970G>A; R2310K) was identified in exon 42 of the splicing factor gene PRPC8 in one patient of this family. Diagnostic restriction enzyme digestion of exon 42 amplified from genomic DNA of all family members revealed that the R2310K mutation segregated fully with the disease. The type I phenotype observed in this family is similar to that described for three other RP13 families with mutations in PRPC8.