RRM
2011 Rustbelt RNA Meeting
RRM
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Poster abstracts
Abstract:
The vertebrate heart develops from a simple, straight, “primary heart tube” to a heart consisting of ventricles, atria, septa, and valves. An important aspect of developmental gene regulation is alternative splicing. In vertebrates, where an increasing number of genes are multi-exonic, alternative splicing can give rise to multiple mature mRNA species from a single precursor mRNA. CELF1 (CUG-BP1- and ETR3-Like Family member 1) is a regulator of alternative splicing. During embryonic development, CELF1 transcript is detected in the myocardial cell layer of the developing heart, while the protein is expressed predominantly in the myocardial nuclei. Protein expression peaks between days 3 and 8, after which protein levels remain low into the postnatal period. These data suggest a temporally and spatially important role for CELF1 in cardiomyocytes during a period in which cardiomyocytes undergo significant waves of proliferation and migration, as part of cardiac morphogenesis.
The objective of this work was to identify ex vivo systems that will allow us to investigate the involvement of CELF1 in early heart development. We can isolate primary cardiomyocytes from the developing heart, knock down CELF1, and investigate molecular interactions identified through biochemical approaches. We are also currently investigating a whole heart culture system, with the aim of observing the progress of developmental events following siRNA knockdown of CELF1 in tissue ex vivo. Current work is focused on optimizing transfection conditions for knockdown and overexpression of CELF1 in the cultured hearts. In addition, biochemical approaches to identify targets of CELF1 splicing regulation activity are being carried out in parallel. These targets will then be investigated using the ex vivo systems described here.
Keywords: CELF1, alternative splicing, heart development
