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

Nature Communications, vol. 12, pp. 5363

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.

Authors 10

1. Sarah Rubin

   Weizmann Institute of Science

   Affiliation as printed

   Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel

2. Ankit Agrawal

   Weizmann Institute of Science

   Affiliation as printed

   Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel

3. Johannes Stegmaier Aachen Institute of Imaging & Computer Vision

   Karlsruhe Institute of Technology · RWTH Aachen University

   Affiliation as printed

   Institute for Automation and Applied Informatics, Karlsruhe Institute of Technology, Karlsruhe, Germany

   Institute of Imaging and Computer Vision, RWTH Aachen University, Aachen, Germany

4. Sharon Krief

   Weizmann Institute of Science

   Affiliation as printed

   Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel

5. Neta Felsenthal

   Weizmann Institute of Science

   Affiliation as printed

   Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel

6. Jonathan Svorai

   Weizmann Institute of Science

   Affiliation as printed

   Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel

7. Yoseph Addadi

   Weizmann Institute of Science

   Affiliation as printed

   Department of Life Science Core Facilities, Weizmann Institute of Science, Rehovot, Israel

8. Paul Villoutreix corresponding

   Aix-Marseille Université · Laboratoire d’Informatique et Systèmes · Institut de Biologie du Développement Marseille

   Affiliation as printed

   LIS (UMR 7020), IBDM (UMR 7288), Turing Center For Living Systems, Aix-Marseille University, Marseille, France. paul.villoutreix@univ-amu.fr

   LIS (UMR 7020), IBDM (UMR 7288), Turing Center For Living Systems, Aix-Marseille University, Marseille, France

9. Tomer Stern corresponding

   Weizmann Institute of Science

   Affiliation as printed

   Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel. tstern@princeton.edu

   Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel

10. Elazar Zelzer corresponding

    Weizmann Institute of Science

    Affiliation as printed

    Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel. Eli.Zelzer@weizmann.ac.il

    Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel