Single-molecule tracking of Nodal and Lefty in live zebrafish embryos supports hindered diffusion model

Timo Kuhn, Amit N. Landge, David Mörsdorf, Jonas Coßmann, Johanna Gerstenecker, Daniel Čapek, Patrick Müller () and J. Christof M. Gebhardt ()
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Timo Kuhn: Ulm University
Amit N. Landge: University of Konstanz
David Mörsdorf: Friedrich Miescher Laboratory of the Max Planck Society
Jonas Coßmann: Ulm University
Johanna Gerstenecker: Ulm University
Daniel Čapek: University of Konstanz
Patrick Müller: University of Konstanz
J. Christof M. Gebhardt: Ulm University

Nature Communications, 2022, vol. 13, issue 1, 1-15

Abstract: Abstract The hindered diffusion model postulates that the movement of a signaling molecule through an embryo is affected by tissue geometry and binding-mediated hindrance, but these effects have not been directly demonstrated in vivo. Here, we visualize extracellular movement and binding of individual molecules of the activator-inhibitor signaling pair Nodal and Lefty in live developing zebrafish embryos using reflected light-sheet microscopy. We observe that diffusion coefficients of molecules are high in extracellular cavities, whereas mobility is reduced and bound fractions are high within cell-cell interfaces. Counterintuitively, molecules nevertheless accumulate in cavities, which we attribute to the geometry of the extracellular space by agent-based simulations. We further find that Nodal has a larger bound fraction than Lefty and shows a binding time of tens of seconds. Together, our measurements and simulations provide direct support for the hindered diffusion model and yield insights into the nanometer-to-micrometer-scale mechanisms that lead to macroscopic signal dispersal.

Date: 2022
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DOI: 10.1038/s41467-022-33704-z

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