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1. Laser ablation and fluid flows reveal the mechanism behind spindle and centrosome positioning

   Wu, H.; Kabacaoğlu, G.; Nazockdast, E.; Chang, H-C; Shelley, M. J.; Needleman, D. J. (January 2024, Nature physics)

   Few techniques are available for studying the nature of forces that drive subcellular dynamics. Here we develop two complementary ones. The first is femtosecond stereotactic laser ablation, which rapidly creates complex cuts of subcellular structures and enables precise dissection of when, where and in what direction forces are generated. The second is an assessment of subcellular fluid flows by comparison of direct flow measurements using microinjected fluorescent nanodiamonds with large-scale fluid-structure simulations of different force transduction models. We apply these techniques to study spindle and centrosome positioning in early Caenorhabditis elegans embryos and to probe the contributions of microtubule pushing, cytoplasmic pulling and cortical pulling upon centrosomal microtubules. Based on our results, we construct a biophysical model to explain the dynamics of centrosomes. We demonstrate that cortical pulling forces provide a general explanation for many behaviours mediated by centrosomes, including pronuclear migration and centration, rotation, metaphase spindle positioning, asymmetric spindle elongation and spindle oscillations. This work establishes methodologies for disentangling the forces responsible for cell biological phenomena. 

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2. The Drosophila mitotic spindle orientation machinery requires activation, not just localization

   https://doi.org/10.15252/embr.202256074  

   Neville, Kathryn E.; Finegan, Tara M.; Lowe, Nicholas; Bellomio, Philip M.; Na, Daxiang; Bergstralh, Dan T. (January 2023, EMBO reports)

   Abstract The orientation of the mitotic spindle at metaphase determines the placement of the daughter cells. Spindle orientation in animals typically relies on an evolutionarily conserved biological machine comprised of at least four proteins – called Pins, Gαi, Mud, and Dynein in flies – that exerts a pulling force on astral microtubules and reels the spindle into alignment. The canonical model for spindle orientation holds that the direction of pulling is determined by asymmetric placement of this machinery at the cell cortex. In most cell types, this placement is thought to be mediated by Pins, and a substantial body of literature is therefore devoted to identifying polarized cues that govern localized cortical enrichment of Pins. In this study we revisit the canonical model and find that it is incomplete. Spindle orientation in theDrosophilafollicular epithelium and embryonic ectoderm requires not only Pins localization but also direct interaction between Pins and the multifunctional protein Discs large. This requirement can be over‐ridden by interaction with another Pins interacting protein, Inscuteable. 

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3. Immune cells use active tugging forces to distinguish affinity and accelerate evolution

   https://doi.org/10.1073/pnas.2213067120  

   Jiang, Hongda; Wang, Shenshen (March 2023, Proceedings of the National Academy of Sciences)

   Cells are known to exert forces to sense their physical surroundings for guidance of motion and fate decisions. Here, we propose that cells might do mechanical work to drive their own evolution, taking inspiration from the adaptive immune system. Growing evidence indicates that immune B cells—capable of rapid Darwinian evolution—use cytoskeletal forces to actively extract antigens from other cells’ surfaces. To elucidate the evolutionary significance of force usage, we develop a theory of tug-of-war antigen extraction that maps receptor binding characteristics to clonal reproductive fitness, revealing physical determinants of selection strength. This framework unifies mechanosensing and affinity-discrimination capabilities of evolving cells: Pulling against stiff antigen tethers enhances discrimination stringency at the expense of absolute extraction. As a consequence, active force usage can accelerate adaptation but may also cause extinction of cell populations, resulting in an optimal range of pulling strength that matches molecular rupture forces observed in cells. Our work suggests that nonequilibrium, physical extraction of environmental signals can make biological systems more evolvable at a moderate energy cost. 

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4. Mechanisms of chromosome biorientation and bipolar spindle assembly analyzed by computational modeling

   https://doi.org/10.7554/eLife.48787  

   Edelmaier, Christopher; Lamson, Adam R; Gergely, Zachary R; Ansari, Saad; Blackwell, Robert; McIntosh, J Richard; Glaser, Matthew A; Betterton, Meredith D (February 2020, eLife)

   The essential functions required for mitotic spindle assembly and chromosome biorientation and segregation are not fully understood, despite extensive study. To illuminate the combinations of ingredients most important to align and segregate chromosomes and simultaneously assemble a bipolar spindle, we developed a computational model of fission-yeast mitosis. Robust chromosome biorientation requires progressive restriction of attachment geometry, destabilization of misaligned attachments, and attachment force dependence. Large spindle length fluctuations can occur when the kinetochore-microtubule attachment lifetime is long. The primary spindle force generators are kinesin-5 motors and crosslinkers in early mitosis, while interkinetochore stretch becomes important after biorientation. The same mechanisms that contribute to persistent biorientation lead to segregation of chromosomes to the poles after anaphase onset. This model therefore provides a framework to interrogate key requirements for robust chromosome biorientation, spindle length regulation, and force generation in the spindle. 

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5. Folding drives cortical thickness variations

   https://doi.org/10.1140/epjst/e2020-000001-6  

   Holland, Maria A.; Budday, Silvia; Li, Gang; Shen, Dinggang; Goriely, Alain; Kuhl, Ellen (November 2020, The European Physical Journal Special Topics)

   null (Ed.)

   Abstract The cortical thickness is a characteristic biomarker for a wide variety of neurological disorders. While the structural organization of the cerebral cortex is tightly regulated and evolutionarily preserved, its thickness varies widely between 1.5 and 4.5 mm across the healthy adult human brain. It remains unclear whether these thickness variations are a cause or consequence of cortical development. Recent studies suggest that cortical thickness variations are primarily a result of genetic effects. Previous studies showed that a simple homogeneous bilayered system with a growing layer on an elastic substrate undergoes a unique symmetry breaking into a spatially heterogeneous system with discrete gyri and sulci. Here, we expand on that work to explore the evolution of cortical thickness variations over time to support our finding that cortical pattern formation and thickness variations can be explained – at least in part – by the physical forces that emerge during cortical folding. Strikingly, as growth progresses, the developing gyri universally thicken and the sulci thin, even in the complete absence of regional information. Using magnetic resonance images, we demonstrate that these naturally emerging thickness variations agree with the cortical folding pattern in n = 9 healthy adult human brains, in n = 564 healthy human brains ages 7–64, and in n = 73 infant brains scanned at birth, and at ages one and two. Additionally, we show that cortical organoids develop similar patterns throughout their growth. Our results suggest that genetic, geometric, and physical events during brain development are closely interrelated. Understanding regional and temporal variations in cortical thickness can provide insight into the evolution and causative factors of neurological disorders, inform the diagnosis of neurological conditions, and assess the efficacy of treatment options. 

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