RESUMEN
A new screening method was developed for the detection of CAG expanded alleles in patients with hereditary ataxia using polymerase chain reaction-based microtiter plate hybridization (PCR-MPH). The system can be applied to detect pathologic alleles by hybridization with the immobilized (CAG)48 repeat probe derived from the unrelated gene 'ERDA1' except for the CAG repeats. We examined 10 individuals with SCA3, 10 with Huntington disease and 30 normal controls (31 controls for SCA3) using this method. The results showed that a clear discrimination was possible in all cases. We suggest that this system be made available for mass screening of patients with hereditary ataxia disorders. This report is the first to demonstrate that a PCR-MPH system can be successfully applied to DNA size differentiation in addition to base pair mismatches. Also, our design of the probe is unique in that the probe motif stem from the unrelated gene sequence and not from the synthetic oligonucleotides.
Asunto(s)
Degeneraciones Espinocerebelosas/diagnóstico , Repeticiones de Trinucleótidos , Alelos , Ataxina-3 , Sondas de ADN , Desoxirribonucleasas de Localización Especificada Tipo II/metabolismo , Pruebas Genéticas , Humanos , Enfermedad de Huntington/diagnóstico , Enfermedad de Huntington/genética , Enfermedad de Machado-Joseph/diagnóstico , Enfermedad de Machado-Joseph/genética , Proteínas del Tejido Nervioso/genética , Proteínas Nucleares , Reacción en Cadena de la Polimerasa , Proteínas Represoras , Sensibilidad y Especificidad , Degeneraciones Espinocerebelosas/genéticaRESUMEN
BACKGROUND: Trinucleotide repetition combined with variable penetrance of expression could be responsible for the complex transmission pattern observed in bipolar affective disorder (BPAD). The purpose of this study was to investigate the association of excess longer allele of KCNN3 and CTG18.1 in the patients with BPAD. METHODS: CAG/CTG repeat distribution in KCNN3, CTG 18.1 and ERDA1 was examined and the copy number of ligation product in repeat expansion detection (RED) was measured in Korean bipolar patients in comparison to ethnically matched healthy controls. RESULTS: No significant difference was found in the allele distribution of those repeats between bipolar patients and controls. Ligation product size in RED was not increased in bipolar patients. However, the copy number of ligation product in RED was highly correlated with CAG/CTG copies of ERDA1 (P=0.0001), partly with CTG 18.1 (P=0.04), but not with KCNN3. CONCLUSIONS: A longer CAG repeat alleles of KCNN3 or CTG 18.1 may not be a risk factor for BPAD in Korean population and the copy number of ligation product in RED in the patients with BPAD is influenced by the longer allele of CAG/CTG of ERDA1 or CTG 18.1.
Asunto(s)
Trastorno Bipolar/genética , Canales de Potasio Calcio-Activados , Canales de Potasio/genética , Expansión de Repetición de Trinucleótido , Repeticiones de Trinucleótidos , Adulto , Alelos , Trastorno Bipolar/clasificación , Trastorno Bipolar/diagnóstico , Femenino , Frecuencia de los Genes/genética , Predisposición Genética a la Enfermedad/genética , Genética de Población , Genotipo , Humanos , Corea (Geográfico) , Masculino , Escalas de Valoración Psiquiátrica , Factores de Riesgo , Canales de Potasio de Pequeña Conductancia Activados por el CalcioRESUMEN
BACKGROUND: Mutations in the FBN1 gene, encoding fibrillin-1, result in Marfan syndrome (MFS). According to previous reports, the mutations in FBN1 share certain characteristics in each family with variable penetrance and overlapping symptoms, even in the same genotype. In the present study, we report six novel mutations and evaluate the clinical significance of these nucleotide changes. METHODS: To screen for nucleotide changes in all 65 exons of the FBN1 gene in 38 unrelated Korean patients, we performed polymerase chain reaction, single-strand conformational polymorphism (SSCP) and sequencing for the shift of the band in SSCP. RESULTS: We identified six mutations: a 2253 del 7 b.p., N1043S, C1254S, L1421F, C1895R and S2662P. CONCLUSIONS: These results suggest that many different mutations are responsible for MFS in the Korean population.
Asunto(s)
Síndrome de Marfan/genética , Proteínas de Microfilamentos/genética , Mutación Puntual , Adolescente , Adulto , Niño , Preescolar , Fibrilina-1 , Fibrilinas , Mutación del Sistema de Lectura , Humanos , Corea (Geográfico) , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Polimorfismo GenéticoRESUMEN
Autosomal dominant cerebellar ataxias (ADCAs) are a heterogeneous group of disorders characterized by degenerative symptoms in the cerebellum, spinal cord, and brain stem. Six different genes have been reported to be associated with ADCA, and the length of trinucleotide repeats of these genes is correlated with the age at onset and severity of symptoms. Although there are strong hereditary effects in these disorders, most of the studies carried out in heterogeneous populations and in small groups obscure the true incidence of these diseases. We examined the frequency of six types of ADCAs in 87 unrelated Korean patients with progressive ataxia and compared the results to the frequencies in other ethnic groups. Spinocerebellar ataxia (SCA) type 2 was the most frequent hereditary ataxia (12.6%) and types 3 and 6 accounted for 4.6% and 6.9% of ataxia patients, respectively. Dentatorubral pallidoluysian atrophy was also found in three patients (3.4%). No instances of SCA types 1 or 7 were detected. These findings show the striking contrast to the white population and a difference from Japanese findings. Our results demonstrate that dentatorubral pallidoluysian atrophy should be included in the differential diagnosis of Korean patients with spinocerebellar ataxia, and that there are strong hereditary effects in patients with ADCAs.
Asunto(s)
Frecuencia de los Genes/genética , Degeneraciones Espinocerebelosas/genética , Expansión de Repetición de Trinucleótido/genética , Estudios de Casos y Controles , Humanos , Corea (Geográfico)/etnología , Degeneraciones Espinocerebelosas/diagnósticoRESUMEN
McCune-Albright syndrome (MAS) is a sporadic disease characterized by café-au-lait spots, polyostotic fibrous dysplasia and hyperfunctional endocrinopathies. To elucidate the mechanism of skin pigmentation, melanocytes, keratinocytes and fibroblasts were primary cultured from the café-au-lait spot of a MAS patient. Then, mutational analysis and morphologic evaluation were performed. Also, cAMP level and tyrosinase gene expression in cultured cells were determined. Only Gsalpha mutation was found in affected melanocytes and the cAMP level in affected melanocytes was higher than that of normal melanocytes. The mRNA expression of tyrosinase gene was increased in the affected melanocytes. This study suggests that skin pigmentation of MAS results from activating mutation of Gsalpha in melanocytes and the mechanism involves the c-AMP-mediated tyrosinase gene activation.