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Tracking intracellular forces and mechanical property changes in mouse one-cell embryo development

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Abstract

Cells comprise mechanically active matter that governs their functionality, but intracellular mechanics are difficult to study directly and are poorly understood. However, injected nanodevices open up opportunities to analyse intracellular mechanobiology. Here, we identify a programme of forces and changes to the cytoplasmic mechanical properties required for mouse embryo development from fertilization to the first cell division. Injected, fully internalized nanodevices responded to sperm decondensation and recondensation, and subsequent device behaviour suggested a model for pronuclear convergence based on a gradient of effective cytoplasmic stiffness. The nanodevices reported reduced cytoplasmic mechanical activity during chromosome alignment and indicated that cytoplasmic stiffening occurred during embryo elongation, followed by rapid cytoplasmic softening during cytokinesis (cell division). Forces greater than those inside muscle cells were detected within embryos. These results suggest that intracellular forces are part of a concerted programme that is necessary for development at the origin of a new embryonic life.

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Fig. 1: Fabricated nanodevices as intracellular sensors.
Fig. 2: ‘H-comb’ nanodevices detect mechanical loads inside mouse zygotes.
Fig. 3: Mechanics during the PM phase.
Fig. 4: Dynamic F-actin redistribution in mouse zygotes.
Fig. 5: Morphological and cytoskeletal changes during elongation and division.
Fig. 6: Tracking perturbation to the mechanical programme of mouse embryos.

Data availability

Supporting data are available in the Source Data files and from the corresponding authors upon reasonable request.

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Acknowledgements

We are grateful to B. Nichols, J. Campos, T. Suárez and C. Tickle for their constructive comments during manuscript preparation and thank Animal Facility support staff for ensuring the welfare of animals used in this work. We are deeply indebted to the late Dino Sharma for help with image collection. We acknowledge support to A.C.F.P. from the Medical Research Council, UK (grant nos. G1000839, MR/N000080/1 and MR/N020294/1) and the Biology and Biological Science Research Council, UK (grant no. BB/P009506/1) and to J.A.P from the Spanish Government, grant nos. TEC2014-51940-C2 and TEC2017-85059-C3 with Feder funding. We also thank the clean room staff of IMB-CNM for assistance with chip fabrication.

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Authors

Contributions

A.C.F.P. and J.A.P. conceived core experiments. M.D., R.G.-M. and J.A.P. performed nanodevice fabrication, T.S. and M.A. performed microinjection and embryo production and M.A. and M.D.V. undertook image collection. Data analysis and modelling were by N.T., M.A., M.I.A., R.C. and J.A.P. A.C.F.P. and J.A.P. wrote most of the manuscript with contributions from the other authors.

Corresponding authors

Correspondence to José Antonio Plaza or Anthony C. F. Perry.

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Supplementary information

Supplementary Information

Supplementary Figs. 1–17, Videos 1–9, discussion, methods and references.

Reporting Summary

Supplementary Video 1

Animation of the axis of rotation of the nanodevice.

Supplementary Video 2

Brightfield video of mouse zygote during PM.

Supplementary Video 3

Brightfield video of mouse zygote containing microsphere

Supplementary Video 4

Tracking particle migration within an embryo.

Supplementary Video 5

Dynamic surface plot of Utr-mCherry in mouse embryos.

Supplementary Video 6

Fluorescence in mouse embryos containing Utr-mCherry.

.Supplementary Video 7

Brightfield video of mouse embryo containing 25-nm ‘H-comb’ nanodevice

Supplementary Video 8

Brightfield video of ‘scary spider’ mouse embryo containing 25-nm ‘H-comb’ nanodevice

Supplementary Video 9

The effect of actomyosin inhibition is consistent with the GES model.

Source data

Source Data Fig. 1

Statistical source data

Source Data Fig. 2

Statistical source data

Source Data Fig. 3

Statistical source data

Source Data Fig. 4

Statistical source data

Source Data Fig. 5

Statistical source data

Source Data Fig. 6

Statistical source data

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Duch, M., Torras, N., Asami, M. et al. Tracking intracellular forces and mechanical property changes in mouse one-cell embryo development. Nat. Mater. 19, 1114–1123 (2020). https://doi.org/10.1038/s41563-020-0685-9

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