Application of 3D MAPs pipeline identifies the morphological sequence chondrocytes undergo and the regulatory role of GDF5 in this process

Sarah Rubin, Ankit Agrawal, Johannes Stegmaier, Sharon Krief, Neta Felsenthal, Jonathan Svorai, Yoseph Addadi, Paul Villoutreix (), Tomer Stern () and Elazar Zelzer ()
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Sarah Rubin: Weizmann Institute of Science
Ankit Agrawal: Weizmann Institute of Science
Johannes Stegmaier: RWTH Aachen University
Sharon Krief: Weizmann Institute of Science
Neta Felsenthal: Weizmann Institute of Science
Jonathan Svorai: Weizmann Institute of Science
Yoseph Addadi: Weizmann Institute of Science
Paul Villoutreix: Aix-Marseille University
Tomer Stern: Weizmann Institute of Science
Elazar Zelzer: Weizmann Institute of Science

Nature Communications, 2021, vol. 12, issue 1, 1-16

Abstract: Abstract The activity of epiphyseal growth plates, which drives long bone elongation, depends on extensive changes in chondrocyte size and shape during differentiation. Here, we develop a pipeline called 3D Morphometric Analysis for Phenotypic significance (3D MAPs), which combines light-sheet microscopy, segmentation algorithms and 3D morphometric analysis to characterize morphogenetic cellular behaviors while maintaining the spatial context of the growth plate. Using 3D MAPs, we create a 3D image database of hundreds of thousands of chondrocytes. Analysis reveals broad repertoire of morphological changes, growth strategies and cell organizations during differentiation. Moreover, identifying a reduction in Smad 1/5/9 activity together with multiple abnormalities in cell growth, shape and organization provides an explanation for the shortening of Gdf5 KO tibias. Overall, our findings provide insight into the morphological sequence that chondrocytes undergo during differentiation and highlight the ability of 3D MAPs to uncover cellular mechanisms that may regulate this process.

Date: 2021
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DOI: 10.1038/s41467-021-25714-0

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