Vascular-specific translational profiling in zebrafish

Wednesday, October 10, 2012 — Poster Session II
Noon – 2:00 p.m	Natcher Conference Center, Building 45	NICHD	DEV-10

Authors

• KF Monzo
• BM Weinstein

Abstract

Vascular development involves regulation of many processes including acquisition of endothelial cell fate, patterning of vessel tracts, and formation of a closed circulatory system, all of which require integration of many different signaling pathways. A number of genes functioning in the endothelium have been identified, but how these genes interact in different pathways and the identity of their effector genes is still largely unknown. To help address this problem, we have been working to develop an unbiased and coprehensive approach to analyze in vivo vascular gene expression. To allow for non-invasive querying of endothelial gene expression, we are adapting a recently described technique termed Translating Ribosome Affinity Purification (TRAP) for use in zebrafish. Studies in mice have demonstrated that transgene-driven expression of an epitope-tagged ribosomal protein subunit can be used to affinity purify and profile mRNAs actively translated within specific cell types in vivo. We have generated “RiboTag” transgenic zebrafish expressing an epitope tagged ribosomal subunit in endothelium under the control of a vascular-specific promoter and have established procedures for purifying polysome-associated RNAs from transgenic embryos. Our RiboTag transgenic fish and the TRAP-RNAseq method provide a powerful new tool for in vivo profiling of vascular gene expression.

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