Canalization of Gene Expression in the Drosophila Blastoderm by Gap Gene Cross Regulation

Abstract

Developing embryos exhibit a robust capability to reduce phenotypic variations that occur naturally or as a result of experimental manipulation. This reduction in variation occurs by an epigenetic mechanism called canalization, a phenomenon which has resisted understanding because of a lack of necessary molecular data and of appropriate gene regulation models. In recent years, quantitative gene expression data have become available for the segment determination process in the Drosophila blastoderm, revealing a specific instance of canalization. These data show that the variation of the zygotic segmentation gene expression patterns is markedly reduced compared to earlier levels by the time gastrulation begins, and this variation is significantly lower than the variation of the maternal protein gradient Bicoid. We used a predictive dynamical model of gene regulation to study the effect of Bicoid variation on the downstream gap genes. The model correctly predicts the reduced variation of the gap gene expression patterns and allows the characterization of the canalizing mechanism. We show that the canalization is the result of specific regulatory interactions among the zygotic gap genes. We demonstrate the validity of this explanation by showing that variation is increased in embryos mutant for two gap genes, Krüppel and knirps, disproving competing proposals that canalization is due to an undiscovered morphogen, or that it does not take place at all. In an accompanying article in PLoS Computational Biology (doi: 10.1371/journal.pcbi.1000303), we show that cross regulation between the gap genes causes their expression to approach dynamical attractors, reducing initial variation and providing a robust output. These results demonstrate that the Bicoid gradient is not sufficient to produce gap gene borders having the low variance observed, and instead this low variance is generated by gap gene cross regulation. More generally, we show that the complex multigenic phenomenon of canalization can be understood at a quantitative and predictive level by the application of a precise dynamical model.

Author Summary

Animals have an astonishing ability to develop reliably in spite of variable conditions during embryogenesis. More than 60 years ago, it was proposed that this property of development, called canalization, results from genetic interactions that adjust biochemical reactions so as to bring about reliable outcomes. Since then, a great deal of progress has been made in understanding the buffering of genotypic and environmental variation, and individual mutations that reveal variation have been identified. However, the mechanisms by which genetic interactions produce canalization are not yet well understood, because this requires molecular data on multiple developmental determinants and models that correctly predict complex interactions. We make use of gene expression data at both high spatial and temporal resolution for the gap genes involved in the segmentation of Drosophila. We also apply a mathematical model to show that cross regulation among the gap genes is responsible for canalization in this system. Furthermore, the model predicted specific interactions that cause canalization, and the prediction was validated experimentally. Our results show that groups of genes can act on one another to reduce variation and highlights the importance of genetic networks in generating robust development.

Abstract

During Drosophila development, the expression patterns of gap genes are much less variable than the Bicoid morphogen gradient. Modeling and experiments show that this specific instance of canalization or developmental robustness occurs by gap gene cross regulation.