Análisis mutacional del gen AMEL en una familia con amelogénesis imperfecta / Mutational Analysis in the AMEL Gene a Family with Amelogenesis Imperfecta

María Claudia Berrocal, Adriana Cavender, Lorenza Jaramillo, Ignacio Briceño, Mónica Melo, Rena D’Souza

Abstract


Mutaciones en los genes que participanen el desarrollo del esmalte dental, como amelogenina (AMEL) y enamelina (ENAM) entre otras, causan amelogénesis imperfecta (AI). OBJETIVO: establecer las mutaciones presentes en el gen AMEL y el modo deherencia en una familia colombiana con AI. MATERIALES Y MÉTODOS: se realizó un pedigree de la familia, evaluación clínica y extracción del ADN de sangre periférica a los individuos participantes. La reacción en cadena de la polimerasa (PCR) fue usada para amplificar los 7 exones del gen AMEL en el brazo corto del cromosoma X y análisis de SSCP fueron realizados en los exones 5 y 6. Electroforesis en gel de poliacrilamida al 6% no denaturantes fueron hechas. Para confirmar ausencia o presencia demutaciones se hizo secuenciamiento de los productos de PCR de los 7 exones. RESULTADOS: el análisis del pedigree mostró un mecanismo de herencia ligado a X y el fenotipo dental observado en los probandos, correspondió a un defecto de tipo hipoplásico. No se identificaron mutaciones en los siete exones del gen amelogenina. CONCLUSIÓN: esta condición tiene gran heterogeneidad genética y posiblemente el fenotipo encontrado esté más relacionado con mutaciones en otros genes concernientes a la formación del esmalte.

 

Mutations of the genes which participate in the enamel development such as amelogenine (AMEL) and enameline (ENAM) among others cause Amelogenesis Imperfecta (A.I). OBJECTIVES: Establish the mutations present in the AMEL gene and the means of inheritage in a Colombian family with A.I. MATERIALS AND METHODS: family pedigree, clinical evaluation and peripheric DNA genomic blood extraction was performed to each of the participating individuals. PCR was used to amplify the 7 exons of the amelogenine gene in the short arm of the X chromosome. SSCP analysis was carried out in the 5 and 6 exons. Gel electrophoresis with non denaturalizing 6% polyacrilamide were carried out to confirm the presence or absence of mutations in the AMEL gene, after which the sequencing of the 7 exon PCR products was performed. RESULTS: the pedigree analysis showed an X heritage linked mechanism. The dental phenotype observed in the proband corresponded to a hypoplastic type defect. There were no identified mutations at a molecular level in the 7 exons of the amelogenine gene. CONCLUSION: this condition has great genetic heterogeneticity and possibly the phenotype found is more related with mutations in other genes linked to the formation of the dental enamel.


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