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Spatiotemporal Mechanical Variation Reveals Critical Role for Rho Kinase During Primitive Streak Morphogenesis

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Abstract

Large-scale morphogenetic movements during early embryo development are driven by complex changes in biochemical and biophysical factors. Current models for amniote primitive streak morphogenesis and gastrulation take into account numerous genetic pathways but largely ignore the role of mechanical forces. Here, we used atomic force microscopy (AFM) to obtain for the first time precise biomechanical properties of the early avian embryo. Our data reveal that the primitive streak is significantly stiffer than neighboring regions of the epiblast, and that it is stiffer than the pre-primitive streak epiblast. To test our hypothesis that these changes in mechanical properties are due to a localized increase of actomyosin contractility, we inhibited actomyosin contractility via the Rho kinase (ROCK) pathway using the small-molecule inhibitor Y-27632. Our results using several different assays show the following: (1) primitive streak formation was blocked; (2) the time-dependent increase in primitive streak stiffness was abolished; and (3) convergence of epiblast cells to the midline was inhibited. Taken together, our data suggest that actomyosin contractility is necessary for primitive streak morphogenesis, and specifically, ROCK plays a critical role. To better understand the underlying mechanisms of this fundamental process, future models should account for the findings presented in this study.

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Acknowledgments

This work was supported by NSF Grant #1000604.

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The authors have no conflict of interest to report.

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Authors and Affiliations

  1. Department of Mechanical Engineering, Georgia Institute of Technology, 315 Ferst Dr. NW, Atlanta, GA, 30332, USA

    Julia Henkels, Jaeho Oh, Wenwei Xu, Drew Owen, Todd Sulchek & Evan Zamir

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  1. Julia Henkels
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  2. Jaeho Oh
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  3. Wenwei Xu
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  4. Drew Owen
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  6. Evan Zamir
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Correspondence to Evan Zamir.

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Associate Editor Eric M. Darling oversaw the review of this article.

Wenwei Xu and Drew Owen contributed equally to this work.

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Henkels, J., Oh, J., Xu, W. et al. Spatiotemporal Mechanical Variation Reveals Critical Role for Rho Kinase During Primitive Streak Morphogenesis. Ann Biomed Eng 41, 421–432 (2013). https://doi.org/10.1007/s10439-012-0652-y

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  • DOI: https://doi.org/10.1007/s10439-012-0652-y

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