Contractile ring composition dictates kinetics of in silico contractility

Constriction kinetics of the cytokinetic ring are expected to depend on dynamic adjustment of ring composition, but the impact of component abundance dynamics on ring constriction is understudied. Computational models generally assume that contractile networks maintain constant composition. To test how compositional dynamics affect constriction kinetics, we first measured F-actin, non-muscle myosin II, septin, and anillin during C. elegans zygotic mitosis. A custom microfluidic device that positioned the cell with the division plane parallel to a light sheet allowed even illumination of the cytokinetic ring. Measured component abundances were implemented in an agent-based 3D model of a membrane-associated contractile ring. With constant network composition, constriction occurred with biologically unrealistic kinetics. Measured changes in component quantities elicited realistic constriction kinetics. Simulated networks were more sensitive to changes in motor and filament amounts, than that of crosslinkers and tethers. Our findings highlight the importance of network composition for actomyosin contraction kinetics. Summary We created a microfluidic device compatible with high numerical aperture light sheet microscopy to measure cytokinetic ring component abundance in the C. elegans zygote. Implementing measured dynamics into our three-dimensional agent-based model of a contractile ring elicited biologically realistic kinetics.

[1]  G. Zachos,et al.  The Abscission Checkpoint: A Guardian of Chromosomal Stability , 2021, Cells.

[2]  R. Gassmann,et al.  Plastin and spectrin cooperate to stabilize the actomyosin cortex during cytokinesis , 2021, Current Biology.

[3]  F. Nédélec,et al.  Bond type and discretization of non-muscle myosin II are critical for simulated contractile dynamics , 2019, bioRxiv.

[4]  D. Sherwood,et al.  Endogenous expression of UNC-59/Septin in C. elegans , 2019, microPublication. Biology.

[5]  R. Gassmann,et al.  Crosslinking activity of non-muscle myosin II is not sufficient for embryonic cytokinesis in C. elegans , 2019, Development.

[6]  T. Pollard,et al.  Molecular Mechanism of Cytokinesis. , 2019, Annual review of biochemistry.

[7]  Linda Z. Shi,et al.  A positive-feedback-based mechanism for constriction rate acceleration during cytokinesis in Caenorhabditis elegans , 2018, eLife.

[8]  Bob Goldstein,et al.  LITE microscopy: Tilted light-sheet excitation of model organisms offers high resolution and low photobleaching , 2018, The Journal of cell biology.

[9]  M. Glotzer,et al.  Spatiotemporal Regulation of RhoA during Cytokinesis , 2018, Current Biology.

[10]  F. Nédélec,et al.  Cross-linkers both drive and brake cytoskeletal remodeling and furrowing in cytokinesis , 2017, bioRxiv.

[11]  G. Jensen,et al.  Coarse-grained simulations of actomyosin rings point to a nodeless model involving both unipolar and bipolar myosins , 2017, bioRxiv.

[12]  A. Nagy,et al.  Bipolar filaments of human nonmuscle myosin 2-A and 2-B have distinct motile and mechanical properties , 2017, bioRxiv.

[13]  B. O’Shaughnessy,et al.  A node organization in the actomyosin contractile ring generates tension and aids stability , 2017, Molecular biology of the cell.

[14]  M. Leptin,et al.  A theory that predicts behaviors of disordered cytoskeletal networks , 2017, bioRxiv.

[15]  François Nédélec,et al.  Plastin increases cortical connectivity to facilitate robust polarization and timely cytokinesis , 2017, The Journal of cell biology.

[16]  J. Yates,et al.  A Sterile 20 Family Kinase and Its Co-factor CCM-3 Regulate Contractile Ring Proteins on Germline Intercellular Bridges , 2017, Current Biology.

[17]  Patrick M. Irwin,et al.  The ultrastructural organization of actin and myosin II filaments in the contractile ring: new support for an old model of cytokinesis , 2017, Molecular biology of the cell.

[18]  Roger D. Kamm,et al.  Morphological Transformation and Force Generation of Active Cytoskeletal Networks , 2017, PLoS Comput. Biol..

[19]  Serge Dmitrieff,et al.  Balance of microtubule stiffness and cortical tension determines the size of blood cells with marginal band across species , 2016, Proceedings of the National Academy of Sciences.

[20]  S. Grill,et al.  Cortical flow aligns actin filaments to form a furrow , 2016, eLife.

[21]  Anne Wald,et al.  Still and rotating myosin clusters determine cytokinetic ring constriction , 2016, Nature Communications.

[22]  J. Dorn,et al.  A theoretical model of cytokinesis implicates feedback between membrane curvature and cytoskeletal organization in asymmetric cytokinetic furrowing , 2016, Molecular biology of the cell.

[23]  F. Nédélec,et al.  Architecture and Connectivity Govern Actin Network Contractility , 2016, Current Biology.

[24]  T. Davies,et al.  Cortical PAR polarity proteins promote robust cytokinesis during asymmetric cell division , 2016, The Journal of cell biology.

[25]  A. Mogilner,et al.  A Combination of Actin Treadmilling and Cross-Linking Drives Contraction of Random Actomyosin Arrays. , 2015, Biophysical journal.

[26]  J. Hammer,et al.  Myosin II isoform co-assembly and differential regulation in mammalian systems. , 2015, Experimental cell research.

[27]  J. Auwerx,et al.  An automated microfluidic platform for C. elegans embryo arraying, phenotyping, and long-term live imaging , 2015, Scientific Reports.

[28]  Jonathan B. Alberts,et al.  Isoforms Confer Characteristic Force Generation and Mechanosensation by Myosin II Filaments. , 2015, Biophysical journal.

[29]  A. Dernburg,et al.  Direct Visualization Reveals Kinetics of Meiotic Chromosome Synapsis. , 2015, Cell reports.

[30]  F. Jülicher,et al.  Dynamic force balances and cell shape changes during cytokinesis. , 2015, Physical review letters.

[31]  Shalin B. Mehta,et al.  Septin assemblies form by diffusion-driven annealing on membranes , 2014, Proceedings of the National Academy of Sciences.

[32]  Basile Audoly,et al.  Furrow constriction in animal cell cytokinesis. , 2014, Biophysical journal.

[33]  J. Sellers,et al.  Characterization of Three Full-length Human Nonmuscle Myosin II Paralogs* , 2013, The Journal of Biological Chemistry.

[34]  K. Verbrugghe,et al.  Condensin and the Spindle Midzone Prevent Cytokinesis Failure Induced by Chromatin Bridges in C. elegans Embryos , 2013, Current Biology.

[35]  E. Homsher,et al.  Kinetic Characterization of Nonmuscle Myosin IIB at the Single Molecule Level* , 2012, The Journal of Biological Chemistry.

[36]  Jay R. Unruh,et al.  Actin depolymerization drives actomyosin ring contraction during budding yeast cytokinesis. , 2012, Developmental cell.

[37]  J. Sellers,et al.  Nonmuscle myosin II exerts tension but does not translocate actin in vertebrate cytokinesis , 2012, Proceedings of the National Academy of Sciences.

[38]  G. Salbreux,et al.  Hydrodynamics of cellular cortical flows and the formation of contractile rings. , 2009, Physical review letters.

[39]  Karen Oegema,et al.  Structural Memory in the Contractile Ring Makes the Duration of Cytokinesis Independent of Cell Size , 2009, Cell.

[40]  Dimitrios Vavylonis,et al.  Assembly Mechanism of the Contractile Ring for Cytokinesis by Fission Yeast , 2008, Science.

[41]  F. Nédélec,et al.  Collective Langevin dynamics of flexible cytoskeletal fibers , 2007, 0903.5178.

[42]  Mark Bachman,et al.  Photoresist with low fluorescence for bioanalytical applications. , 2007, Analytical chemistry.

[43]  E. Munro,et al.  Astral Signals Spatially Bias Cortical Myosin Recruitment to Break Symmetry and Promote Cytokinesis , 2007, Current Biology.

[44]  J. Sellers,et al.  Load-dependent mechanism of nonmuscle myosin 2 , 2007, Proceedings of the National Academy of Sciences.

[45]  K. Oegema,et al.  Anillin and the septins promote asymmetric ingression of the cytokinetic furrow. , 2007, Developmental cell.

[46]  William H Guilford,et al.  Mechanics of actomyosin bonds in different nucleotide states are tuned to muscle contraction. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[47]  Denis Wirtz,et al.  Probing single-cell micromechanics in vivo: the microrheology of C. elegans developing embryos. , 2006, Biophysical journal.

[48]  S. Yonemura,et al.  Centralspindlin regulates ECT2 and RhoA accumulation at the equatorial cortex during cytokinesis , 2006, Journal of Cell Science.

[49]  Xiaodong Wu,et al.  Optimal Surface Segmentation in Volumetric Images-A Graph-Theoretic Approach , 2006, IEEE Transactions on Pattern Analysis and Machine Intelligence.

[50]  Thomas D Pollard,et al.  Counting Cytokinesis Proteins Globally and Locally in Fission Yeast , 2005, Science.

[51]  G. von Dassow,et al.  A microtubule-dependent zone of active RhoA during cleavage plane specification , 2005, The Journal of cell biology.

[52]  A. Straight,et al.  Anillin binds nonmuscle myosin II and regulates the contractile ring. , 2004, Molecular biology of the cell.

[53]  J. Priess,et al.  Cortical flows powered by asymmetrical contraction transport PAR proteins to establish and maintain anterior-posterior polarity in the early C. elegans embryo. , 2004, Developmental cell.

[54]  Davi Geiger,et al.  Segmentation by grouping junctions , 1998, Proceedings. 1998 IEEE Computer Society Conference on Computer Vision and Pattern Recognition (Cat. No.98CB36231).

[55]  D. Capco,et al.  Analysis of cellular signaling events, the cytoskeleton, and spatial organization of macromolecules during early Xenopus development. , 1991, Methods in cell biology.

[56]  T D Pollard,et al.  Rate constants for the reactions of ATP- and ADP-actin with the ends of actin filaments , 1986, The Journal of cell biology.

[57]  J. White,et al.  On the mechanisms of cytokinesis in animal cells. , 1983, Journal of theoretical biology.

[58]  Y. Hiramoto,et al.  FORCE EXERTED BY THE CLEAVAGE FURROW OF SEA URCHIN EGGS , 1975, Development, growth & differentiation.

[59]  K. Dan,et al.  Tension at the surface of the dividing sea-urchin egg. , 1972, The Journal of experimental biology.

[60]  T. E. Schroeder The contractile ring. II. Determining its brief existence, volumetric changes, and vital role in cleaving Arbacia eggs. , 1972 .