Modeling of the Dorsal Gradient across Species Reveals Interaction between Embryo Morphology and Toll Signaling Pathway during Evolution

Morphogenetic gradients are essential to allocate cell fates in embryos of varying sizes within and across closely related species. We previously showed that the maternal NF-κB/Dorsal (Dl) gradient has acquired different shapes in Drosophila species, which result in unequally scaled germ layers along the dorso-ventral axis and the repositioning of the neuroectodermal borders. Here we combined experimentation and mathematical modeling to investigate which factors might have contributed to the fast evolutionary changes of this gradient. To this end, we modified a previously developed model that employs differential equations of the main biochemical interactions of the Toll (Tl) signaling pathway, which regulates Dl nuclear transport. The original model simulations fit well the D. melanogaster wild type, but not mutant conditions. To broaden the applicability of this model and probe evolutionary changes in gradient distributions, we adjusted a set of 19 independent parameters to reproduce three quantified experimental conditions (i.e. Dl levels lowered, nuclear size and density increased or decreased). We next searched for the most relevant parameters that reproduce the species-specific Dl gradients. We show that adjusting parameters relative to morphological traits (i.e. embryo diameter, nuclear size and density) alone is not sufficient to reproduce the species Dl gradients. Since components of the Tl pathway simulated by the model are fast-evolving, we next asked which parameters related to Tl would most effectively reproduce these gradients and identified a particular subset. A sensitivity analysis reveals the existence of nonlinear interactions between the two fast-evolving traits tested above, namely the embryonic morphological changes and Tl pathway components. Our modeling further suggests that distinct Dl gradient shapes observed in closely related melanogaster sub-group lineages may be caused by similar sequence modifications in Tl pathway components, which are in agreement with their phylogenetic relationships.Author Summary: Embryo size can vary greatly among closely related species. How tissue specification either scales or is modified in the developing embryo in different species is an ongoing investigation in developmental biology. Here we asked how embryo morphology and specific molecular pathways influence tissue specification by altering the distribution of morphogens. Morphogens are molecules that form gradients that regulate gene expression patterns in a dosage-dependent fashion that result in tissue specification, and therefore are a prime target for evolution in order to adjust or maintain tissue proportions in relation to overall embryo size. We used a mathematical model to identify factors that influence the distribution of the Dorsal morphogen gradient that is responsible for patterning the dorsal-ventral axis of the Drosophila fruit fly embryo. We obtained experimental data from mutant conditions and different species of Drosophila to calibrate our model and found an interaction between embryo morphology and regulation of the Toll pathway, which regulates the Dorsal gradient. Furthermore, the model predicts that closely related species share similar modifications in Toll pathway components resulting in their species-specific gradient shapes, which are supported by interspecies amino acid comparison of the components Dorsal and Cactus.