PgmNr D1202: Organ-Specific Transcriptional Co-regulation of Growth and Form in the Drosophila Embryo.

Authors:
R. Loganathan 1 ; J. Lee 2 ; M. B. Wells 1 ; E. Grevengoed 1 ; M. Slattery 2 ; D. Andrew 1


Institutes
1) Johns Hopkins University, Baltimore, MD; 2) University of Minnesota.


Keyword: ectodermal derivatives

Abstract:

Tubular organogenesis is critically dependent on the coordinate regulation of growth and form as the size and shape of tubular tissues are the prime determinants of their functional complexity. Transcription factors, which regulate the spatiotemporal patterns of gene expression during development, affect multiple aspects of organogenesis. In this work, we describe the distinct role of Ribbon (Rib) during organogenesis of epithelial tubes. Our investigations of the embryonic salivary gland (unbranched tubular organ) and the trachea (branched tubular organ) demonstrate that Rib co-regulates growth and form during tubular organogenesis. In rib null embryos, we observed remarkable changes in cell shape and size of both organs, which underlie the characteristic tube elongation defects. Notably, the changes in growth and form in the rib mutants occurred without effects on cell specification, proliferation, apoptosis, apicobasal polarity or junctional integrity. How does Rib selectively co-regulate growth and form in these tubular epithelia?  To identify the transcriptional targets of Rib, we performed whole embryo microarray gene expression analysis comparing RNA samples (1.5X minimal fold change, P < 0.05) from age-matched wild-type and rib null embryos. Microarray gene expression analysis showed that 774 genes were potentially activated whereas 1176 genes were potentially repressed by Rib. To distinguish direct tissue-specific Rib targets, we performed ChIP-seq analysis in embryos driving rib expression specifically in the salivary gland and the trachea. Superposition of potential Rib targets obtained from the microarray gene expression and ChIP-seq analyses resulted in a total of 60 genes (20 activated, 40 repressed) subject to direct Rib regulation in the salivary gland. Interestingly, Rib was subject to transcriptional autoregulation during salivary gland formation. Meanwhile, 242 genes (54 activated, 188 repressed) were subject to potential direct Rib regulation in the trachea. A subset of potential Rib targets in both organs was validated by in situ hybridization and qRT-PCR. Intriguingly, Rib targets in both tubular organs functionally clustered around cell growth and morphogenesis, suggesting that the co-regulation of growth and form is the primary role for Rib during tubular organogenesis. Moreover, the mechanism by which Rib mediates co-regulation of growth and form is organ-specific as revealed by distinct target clusters in the salivary gland (translational machinery) and the trachea (growth factor signaling machinery). Collectively, our results suggest organ-specific mechanisms in the transcriptional co-regulation of growth and form by Rib in the Drosophila embryo.



Flybase Genetic Index:
1. FlyBase gene symbol: Rib; FBgn: FBgn0003254