Extracellular interactions and ligand degradation shape the nodal morphogen gradient

Wang, Yin; Wang, Xi; Wohland, Thorsten; Sampath, Karuna

doi:10.7554/elife.13879

Cited by 57 publications

(77 citation statements)

References 67 publications

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“…We further investigated how the finger 1 domain limits the distribution range of Nodal. While a previous study has suggested that binding of Nodal to the Acvr2b receptor slows down the Nodal diffusion in zebrafish 38 , the overexpression or deletion of Acvr2b in our system did not change the Nodal distribution range (Supplementary fig. 5b).…”

Section: Resultscontrasting

confidence: 67%

Synthetic mammalian pattern formation driven by differential diffusivity of Nodal and Lefty

Sekine

Shibata

Ebisuya

2018

Preprint

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11Pattern formation is fundamental for embryonic development. Although synthetic 12 biologists have created several patterns, a synthetic mammalian reaction-diffusion pattern 13 has yet to be realized. TGF-β family proteins Nodal and Lefty have been proposed to 14 meet the conditions for reaction-diffusion patterning: Nodal is a short-range activator that 15 enhances the expression of Nodal and Lefty whereas Lefty acts as a long-range inhibitor 16 against Nodal. However, the pattern forming possibility of the Nodal-Lefty signaling has 17 never been directly tested, and the underlying mechanisms of differential diffusivity of 18 Nodal and Lefty remain unclear. Here, through a combination of synthetic biology and 19 theoretical modeling, we show that a reconstituted minimal network of the Nodal-Lefty 20 signaling spontaneously gives rise to a pattern in mammalian cell culture. Surprisingly, 21 extracellular Nodal was confined underneath the cells as small clusters, resulting in a 22 narrow distribution range compared with Lefty. We further found that the finger 1 domain 23 of the Nodal protein is responsible for its short-range distribution. By transplanting the 24 finger 1 domain of Nodal into Lefty, we converted the originally long-range distribution 25 of Lefty to a short-range one, successfully preventing the pattern formation. These results 26 indicate that the differences in the localization and domain structures between Nodal and 27 Lefty, combined with the activator-inhibitor topology, are sufficient for reaction-diffusion 28 pattern formation in mammalian cells. 29 30 Main 31One of the goals of synthetic biology is creating a synthetic tissue to understand natural 32 developmental mechanisms 1-3 , to explore the origin of multicellularity 4 and to engineer a 33 programmable tissue for therapeutic purposes 5,6 . The first step towards a synthetic tissue 34 is controlling pattern formation, which enables to place different types of cells properly 35 in a tissue. Several synthetic cellular patterns have been reported previously: Ring patterns were created in genetically engineered bacteria that can sense the concentrations 1 of small molecules 7,8 . In mammalian cells, 2D and 3D patterns were created based on 2 engineered cell sorting mechanisms 9,10 . However, there is another pattern formation 3 mechanism that has not been artificially created in mammalian cells despite its 4 importance: the reaction-diffusion (RD) patterning system. 5The concept of a self-organizing RD system was first proposed by Alan Turing 6 as a chemical system of interacting and diffusible molecules giving rise to various stable 7 patterns, such as spots and stripes [11][12][13][14] . Recent studies have suggested that RD system 8 underlies a number of developmental patterning phenomena, including digit formation in 9 the limb 15,16 , pigmentation on the skin 17 , the formation of hair follicles and feather buds 10 on the skin 18,19 , branching morphogenesis in the lung 20 and rugae formation in the palate 21 . 11In the field of sy...

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Section: Resultscontrasting

confidence: 67%

Synthetic mammalian pattern formation driven by differential diffusivity of Nodal and Lefty

Sekine

Shibata

Ebisuya

2018

Preprint

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“…Our results indicated that Lefty1 expression is induced by Nodal signaling and that Lefty1 restricts the number of Lefty1 + cells, a scenario reminiscent of the self-enhancement and lateral inhibition (SELI) Nodal-Lefty regulatory network that operates during L–R patterning at E8.0 24 . Given that Lefty proteins diffuse faster than Nodal 25 , whether a Nodal-Lefty SELI system operates in peri-implantation embryos will depend on whether Nodal expression is positively regulated by Nodal itself. We therefore investigated how Nodal expression is regulated at the blastocyst stage—in particular, whether it is positively autoregulated as it is at E8.0.…”

Section: Resultsmentioning

confidence: 99%

Both Nodal signalling and stochasticity select for prospective distal visceral endoderm in mouse embryos

2017

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Anterior–posterior (A–P) polarity of mouse embryos is established by distal visceral endoderm (DVE) at embryonic day (E) 5.5. Lefty1 is expressed first at E3.5 in a subset of epiblast progenitor cells (L1epi cells) and then in a subset of primitive endoderm cells (L1dve cells) fated to become DVE. Here we studied how prospective DVE cells are selected. Lefty1 expression in L1epi and L1dve cells depends on Nodal signaling. A cell that experiences the highest level of Nodal signaling begins to express Lefty1 and becomes an L1epi cell. Deletion of Lefty1 alone or together with Lefty2 increased the number of prospective DVE cells. Ablation of L1epi or L1dve cells triggered Lefty1 expression in a subset of remaining cells. Our results suggest that selection of prospective DVE cells is both random and regulated, and that a fixed prepattern for the A–P axis does not exist before the blastocyst stage.

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“…The step size along each axis was generated by random number function ran2 and displacement followed <x 2 > = 2Dt, where D = 60 µm 2 /s is the diffusion coefficient. This is based on the measurement of the diffusion of TGF-β ligand NODAL in zebrafish 31 . The diffusing ligands were not allowed to pass tight junctions, cell membranes without receptors, or the boundaries of pre-amniotic cavity and interstitial space.…”

Section: Methodsmentioning

confidence: 99%

Embryo geometry drives formation of robust signaling gradients through receptor localization

Zhang

Zwick

Loew

et al. 2018

Preprint

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Morphogen signals are essential for cell fate specification during embryogenesis. Some receptors that sense these morphogen signals are known to localize to only the apical or basal membrane of polarized cell lines in vitro. How such localization affects morphogen sensing and patterning in the developing embryo remains unknown. Here, we show in the early mouse embryo that the formation of a robust BMP signaling gradient depends on restricted, basolateral localization of the BMP receptors. Mis-localizing these receptors to apical membrane leads to ectopic BMP signaling in vivo in the mouse embryo. To reach the basolaterally localized receptors in epiblast, BMP4 ligand, secreted by the extra-embryonic ectoderm, has to diffuses through the narrow interstitial space between the epiblast and the underlying endoderm. This restricted, basolateral diffusion creates a signaling gradient. The embryo geometry further buffers the gradient from fluctuations in the levels of BMP4. Our results demonstrate the importance of receptor localization and embryo geometry in shaping morphogen signaling during embryogenesis. 2 The early mouse embryo (E6.0-E6.5) adopts an egg-cylinder geometry ( Fig. 1a) 5,6,16 . It contains a lumen (the pre-amniotic cavity) encased by two epithelial tissues: the epiblast and extraembryonic ectoderm (ExE). The ExE secretes the morphogen BMP4, which is sensed by receptors in epiblast 5,6 . The resulting BMP signaling is required for the differentiation of the epiblast into mesoderm 3,4 . Both the epiblast and ExE have stereotyped epithelial tissue geometries 17 , with their apical membranes surrounding the lumen and their basolateral membranes facing a narrow interstitial space (between these tissues and the underlying visceral endoderm (VE)). This lumen and interstitial space are separated by impermeable tight junctions present throughout the epithelia except at the border between the ExE and epiblast (Fig. 1a). Indeed, when small-molecule dye fluorescein was injected into the pre-amniotic cavity of an E6.5 mouse embryo, it did not penetrate the epiblast or ExE but diffused through a channel at the edge of the epiblast ( Supplementary Fig. 1). Thus, the extracellular space in the embryo through which BMP4 ligands diffuse is compartmentalized into a lumen and an interstitial space. Here, by combining mathematical modeling, quantitative imaging, embryological perturbation, and microfluidics, we demonstrate that restricted receptor localization in conjunction with the embryo geometry, constrains the diffusion of and therefore response to BMP4 ligands. We show that the BMP4 signaling gradient arises from the edge of epiblast even under conditions of uniform BMP4 stimulation. Further, the interplay between restricted receptor localization and the embryo geometry buffers BMP4 ligands in pre-amniotic cavity through an entropic effect. This entropic buffering renders the BMP4 signaling gradient robust to fluctuations in BMP4 level. Consistently, mis-localizing BMP receptors in the mouse embryo leads to ectopic B...

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Extracellular interactions and ligand degradation shape the nodal morphogen gradient

Cited by 57 publications

References 67 publications

Synthetic mammalian pattern formation driven by differential diffusivity of Nodal and Lefty

Synthetic mammalian pattern formation driven by differential diffusivity of Nodal and Lefty

Both Nodal signalling and stochasticity select for prospective distal visceral endoderm in mouse embryos

Embryo geometry drives formation of robust signaling gradients through receptor localization

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