David M. Umulis

Shaping BMP morphogen gradients in the Drosophila embryo and pupal wing

In the early Drosophila embryo, BMP-type ligands act as morphogens to suppress neural induction and to specify the formation of dorsal ectoderm and amnioserosa. Likewise, during pupal wing development, BMPs help to specify vein versus intervein cell fate. Here, we review recent data suggesting that these two processes use a related set of extracellular factors, positive feedback, and BMP heterodimer formation to achieve peak levels of signaling in spatially restricted patterns. Because these signaling pathway components are all conserved, these observations should shed light on how BMP signaling is modulated in vertebrate development.

Facilitated Transport of a Dpp/Scw Heterodimer by Sog/Tsg Leads to Robust Patterning of the Drosophila Blastoderm Embryo

Patterning the dorsal surface of the Drosophila blastoderm embryo requires Decapentaplegic (Dpp) and Screw (Scw), two BMP family members. Signaling by these ligands is regulated at the extracellular level by the BMP binding proteins Sog and Tsg. We demonstrate that Tsg and Sog play essential roles in transporting Dpp to the dorsal-most cells. Furthermore, we provide biochemical and genetic evidence that a heterodimer of Dpp and Scw, but not the Dpp homodimer, is the primary transported ligand and that the heterodimer signals synergistically through the two type I BMP receptors Tkv and Sax. We propose that the use of broadly distributed Dpp homodimers and spatially restricted Dpp/Scw heterodimers produces the biphasic signal that is responsible for specifying the two dorsal tissue types. Finally, we demonstrate mathematically that heterodimer levels can be less sensitive to changes in gene dosage than homodimers, thereby providing further selective advantage for using heterodimers as morphogens.

The extracellular regulation of bone morphogenetic protein signaling.

In many cases, the level, positioning and timing of signaling through the bone morphogenetic protein (BMP) pathway are regulated by molecules that bind BMP ligands in the extracellular space. Whereas many BMP-binding proteins inhibit signaling by sequestering BMPs from their receptors, other BMP-binding proteins cause remarkably context-specific gains or losses in signaling. Here, we review recent findings and hypotheses on the complex mechanisms that lead to these effects, with data from developing systems, biochemical analyses and mathematical modeling.

The BMP-Binding Protein Crossveinless 2 Is a Short-Range, Concentration-Dependent, Biphasic Modulator of BMP Signaling in Drosophila

In Drosophila, the secreted BMP-binding protein Short gastrulation (Sog) inhibits signaling by sequestering BMPs from receptors, but enhances signaling by transporting BMPs through tissues. We show that Crossveinless 2 (Cv-2) is also a secreted BMP-binding protein that enhances or inhibits BMP signaling. Unlike Sog, however, Cv-2 does not promote signaling by transporting BMPs. Rather, Cv-2 binds cell surfaces and heparan sulfate proteoglygans and acts over a short range. Cv-2 binds the type I BMP receptor Thickveins (Tkv), and we demonstrate how the exchange of BMPs between Cv-2 and receptor can produce the observed biphasic response to Cv-2 concentration, where low levels promote and high levels inhibit signaling. Importantly, we show also how the concentration or type of BMP present can determine whether Cv-2 promotes or inhibits signaling. We also find that Cv-2 expression is controlled by BMP signaling, and these combined properties enable Cv-2 to exquisitely tune BMP signaling.

Brat promotes stem cell differentiation via control of a bistable switch that restricts BMP signaling.

Summary Drosophila ovarian germline stem cells (GSCs) are maintained by Dpp signaling and the Pumilio (Pum) and Nanos (Nos) translational repressors. Upon division, Dpp signaling is extinguished, and Nos is downregulated in one daughter cell, causing it to switch to a differentiating cystoblast (CB). However, downstream effectors of Pum-Nos remain unknown, and how CBs lose their responsiveness to Dpp is unclear. Here, we identify Brain Tumor (Brat) as a potent differentiation factor and target of Pum-Nos regulation. Brat is excluded from GSCs by Pum-Nos but functions with Pum in CBs to translationally repress distinct targets, including the Mad and dMyc mRNAs. Regulation of both targets simultaneously lowers cellular responsiveness to Dpp signaling, forcing the cell to become refractory to the self-renewal signal. Mathematical modeling elucidates bistability of cell fate in the Brat-mediated system, revealing how autoregulation of GSC number can arise from Brat coupling extracellular Dpp regulation to intracellular interpretation.

Organism-Scale Modeling of Early Drosophila Patterning via Bone Morphogenetic Proteins

Advances in image acquisition and informatics technology have led to organism-scale spatiotemporal atlases of gene expression and protein distributions. To maximize the utility of this information for the study of developmental processes, a new generation of mathematical models is needed for discovery and hypothesis testing. Here, we develop a data-driven, geometrically accurate model of early Drosophila embryonic bone morphogenetic protein (BMP)-mediated patterning. We tested nine different mechanisms for signal transduction with feedback, eight combinations of geometry and gene expression prepatterns, and two scale-invariance mechanisms for their ability to reproduce proper BMP signaling output in wild-type and mutant embryos. We found that a model based on positive feedback of a secreted BMP-binding protein, coupled with the experimentally measured embryo geometry, provides the best agreement with population mean image data. Our results demonstrate that using bioimages to build and optimize a three-dimensional model provides significant insights into mechanisms that guide tissue patterning.

Mechanisms of scaling in pattern formation

Many organisms and their constituent tissues and organs vary substantially in size but differ little in morphology; they appear to be scaled versions of a common template or pattern. Such scaling involves adjusting the intrinsic scale of spatial patterns of gene expression that are set up during development to the size of the system. Identifying the mechanisms that regulate scaling of patterns at the tissue, organ and organism level during development is a longstanding challenge in biology, but recent molecular-level data and mathematical modeling have shed light on scaling mechanisms in several systems, including Drosophila and Xenopus. Here, we investigate the underlying principles needed for understanding the mechanisms that can produce scale invariance in spatial pattern formation and discuss examples of systems that scale during development.

Robustness of Embryonic Spatial Patterning in Drosophila melanogaster

Publisher Summary Mathematical models of embryonic development can provide insights into the complex interactions between spatial variations in morphogens, signal transduction, and intracellular response in patterning processes. Mechanistic models of specific processes based on our current understanding of them provide an additional tool to help in understanding the complex regulation of development. The chapter discusses a number of different aspects of robustness in Drosophila embryonic patterning, and have shown how the models lead to new insights concerning scale-invariance in AP patterning, the role of network topology and signature in the switching network used for control of the segment polarity genes, and the role of signaling via heterodimers in dorsal–ventral (DV) patterning. The modular decomposition of DV patterning described in the chapter is based on the current understanding of the kinetic interactions between morphogens and inhibitors in the perivitelline (PV) space, and incorporates the various dimeric forms of inhibitors and signaling the bone morphogenetic proteins (BMPs). Scale-invariance is a basic feature of a number of developmental processes. A rather surprising result is that the robustness of DV spatial patterning can essentially be predicted from an analysis of the local dynamics. The analysis of the spatially distributed system corroborates the earlier results and provides further evidence for the selective advantage of using heterodimers for signal transduction and morphogenesis.

Analysis of dynamic morphogen scale invariance

During the development of some tissues, fields of multipotent cells differentiate into distinct cell types in response to the local concentration of a signalling factor called a morphogen. Typically, individual organisms within a population differ in size, but their body plans appear to be scaled versions of a common template. Similarly, closely related species may differ by three or more orders of magnitude in size, yet common structures between species scale to have similar proportions. In standard reaction–diffusion equations, the morphogen range has a length scale that depends on a balance between kinetic and transport processes and not on the length or size of the field of cells being patterned. However, as shown here for a class of morphogen-patterning systems, a number of conditions lead to scale invariance of the morphogen distribution at equilibrium and during the transient approach to equilibrium. Equilibrium scale invariance requires conservation of the total binding site number and total input flux. Dynamic scale invariance additionally requires sufficient binding to slow the diffusion of ligand. The equations derived herein can be extended to the study of other perturbations to gain further insight into the processes regulating the robustness and scaling of morphogen-mediated pattern formation.

Regulation of BMP activity and range in Drosophila wing development.

Bone morphogenetic protein (BMP) signaling controls development and maintenance of many tissues. Genetic and quantitative approaches in Drosophila reveal that ligand isoforms show distinct function in wing development. Spatiotemporal control of BMP patterning depends on a network of extracellular proteins Pent, Ltl and Dally that regulate BMP signaling strength and morphogen range. BMP-mediated feedback regulation of Pent, Ltl, and Dally expression provides a system where cells actively respond to, and modify, the extracellular morphogen landscape to form a gradient that exhibits remarkable properties, including proportional scaling of BMP patterning with tissue size and the modulation of uniform tissue growth. This system provides valuable insights into mechanisms that mitigate the influence of variability to regulate cell-cell interactions and maintain organ function.