IBR   13079
INSTITUTO DE BIOLOGIA MOLECULAR Y CELULAR DE ROSARIO
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
Ribosome Biogenesis in zebrafish development: new approaches to study ribosomopathies
Autor/es:
MOUGUELAR, VALERIA; PORCEL DE PERALTA, MAURO; CARNEVALE, MATIAS; CALCATERRA, NORA; COUX, GABRIELA
Lugar:
Mar del Plata
Reunión:
Congreso; 51 Annual Meeting Argentine Society for Biochemistry and Molecular Biology; 2015
Resumen:
Collectively, ribosomopathies are caused by defects in ribosome biogenesis.Although these disorders encompass deficiencies in a ubiquitous and fundamentalprocess, the clinical manifestations are highly variable and typically displaytissue specificity. An example is the Treacher Collins Syndrome (TCS), acraniofacial development disorder  due tomutations in the TCOF1 gene, which encodes the protein TREACLE. Previous ly wehave developed a zebrafish TCS-like model through the injection of translation-blockingmorpholinos against the TCOF1 ortholog (nolc1). Our aims here were: i) to developanti-nolc1 antibodies to deepen our studies in zebrafish TCS model, ii) to designan efficient and simple method to measure ribosome biosynthesis in TCS-like fish embryos. Zebrafish N-terminalnolc1 was cloned in the pRSET A plasmid, expressed and purified by Ni-agarose chromatography.The rabbit polyclonal antibodies obtained were highly specific and allowed usto corroborate a diminished translation of the homolog of TREACLE. We designedspecific primers for the 5?ETS and ITS1 regions of the 47S pre-rRNA. rRNA transcription was estimated by RT-qPCR at different developmentalstages in normal and TCS-like zebrafish  embryos.Our results highlight the necessity to perform a scan along embryo developmentto better detect the ribosomal biosynthetic performance under apathological condition as TCS