Menu

Close

Planar differential growth rates determine the position of folds in complex epithelia

Melda Tozluoğlu, Maria Duda, Natalie J Kirkland, Ricardo Barrientos, Jemima J Burden, José J Muñoz, Yanlan Mao

Preprint posted on January 09, 2019 https://www.biorxiv.org/content/10.1101/515528v1?rss=1

and

Buckling of epithelium growing under spherical confinement

Anastasiya Trushko, Ilaria Di Meglio, Aziza Merzouki, Carles Blanch-Mercader, Shada Abuhattum, Jochen Guck, Kevin Alessandri, Pierre Nassoy, Karsten Kruse, Bastien Chopard, Aurelien Roux

Preprint posted on January 07, 2019 https://www.biorxiv.org/content/10.1101/513119v1

Grow under compressive forces and you will automatically fold

Selected by Sundar Naganathan

Background

Folded epithelial sheets and tubes are observed across many organs in different phyla. How does a planar epithelial sheet fold into a complex 3D structure? Many different mechanisms such as apical constriction, basal relaxation and lateral constriction have been put forward to explain the mechanisms by which folding occurs during morphogenesis (see introduction of Tozluoglu et al. for an exhaustive list of mechanisms with references). All these processes involve active mechanical forces generated by the cytoskeleton intracellularly, which are communicated across cells in the tissue. Recent studies1-4, however, have demonstrated that passive forces that arise from mechanical instabilities between epithelial tissues and their surroundings can also lead to folding.

In most cases, epithelial tissues in embryos are not present in isolation. They are surrounded by an extracellular matrix (ECM) and/or other tissues that have different material properties. It turns out that if an epithelial tissue experiences compressive forces from its surroundings, the tissue tends to buckle and fold. The compressive forces can be either due to growth of the tissue by cell division confined by the ECM or due to differential growth of two connected tissues where one tissue grows faster than the other.

Irrespective of the mechanism, it is still unknown, how the initial position of the fold is determined. Tozluoglu et al. by investigating Drosophila wing imaginal disc, propose a novel mechanism where planar differential growth under the compression of ECM drives precise positioning of folds. In another preprint, Trushko et al. develop an in vitro model system to study folding of tissues growing under compression.

Key discoveries

Folding in Drosophila wing imaginal disc (Tozluoglu et al.):

The wing imaginal disc forms three folds between the hinge and pouch regions in the columnal layer between 80 and 96 hrs after egg laying. To understand fold formation, the authors first built a computational model where the epithelial tissue was represented as a non-homogeneous continuous material with temporally invariant material properties. When the tissue was allowed to proliferate spatially uniformly, no folds were observed to form in the absence of external resistance. Folds were observed with an introduction of external viscous resistance and correct number of folds (three) were obtained when apical stiffness was also increased concurrently. However, the shapes and positions of folds still could not be reproduced.

The authors then proceeded to experimentally test some of the assumptions in their model. Clonal analysis of the columnal layer revealed that growth is not uniform and that growth is relatively higher in the notum and pouch regions during early stages of folding. Moreover, an electron microscopy study revealed that the basement membrane (BM) gets thicker as folding proceeds. Implementing a differential planar growth rate as well as by considering the basement membrane as an elastic layer in the simulations, the authors show that the precise position, number and morphology of the folds can be predicted both in wild type as well as in wingless mutant scenarios.

Buckling of epithelium in vitro (Trushko et al.):

The authors encapsulated MDCK-II cells in spherical alginate capsules, whose inner surface had a few micrometres thick matrigel to which the cells adhered. The cells were allowed to proliferate thereby forming a monolayer. Incredibly, in this simple set up, after a few hrs of proliferation, folding of the monolayer was observed, where the monolayer detached from the alginate shell and bent inwards. As the monolayer proliferated, the thickness of the elastic alginate shell was observed to reduce suggesting that the cells were exerting pressure on the external environment. Knowing the thickness, Young’s modulus and stiffness of the capsule, monolayer folding was estimated to occur at a pressure of 100 Pa.

The authors then developed a continuum model of their system by considering the monolayer as an elastic ring that is confined in a circular domain. As the radius of the elastic ring increased, a first order buckling transition was observed similar to their experimental results. Furthermore, they performed simulations using a 2D vertex model and show that an effective friction of the monolayer to the matrigel and active cell mechanics are required to reproduce experimentally observed fold morphologies.

Why I chose these preprints

The preprints show through in vitro, in vivo and computational approaches that folding can occur through passive mechanical forces. Importantly, Tozluoglu et al. demonstrate that the position of folds can be explained by considering planar differential growth rate, which provides a crucial add-on for theoretical descriptions of folds. This is likely to motivate researchers investigating other instances of folding in diverse contexts to explore differential planar growth rates as a plausible mechanism. On the other hand, Trushko et al. bring out a much-needed in vitro model to study folding, where the mechanics of the folding process can be explored in depth.

Open questions

Wing imaginal disc:

  1. The authors experimentally measure differential growth rate in the columnal layer of the wing imaginal disc. What triggers this differential growth?
  2. The BM was assumed to be of uniform thickness in their model. Could the BM exhibit differential confinement on the columnar layer, which would bias the folding process to specific locations?
  3. Can the growth rate be changed in a spatiotemporal fashion in the columnal layer? According to their model, this should change the position of folds. Without this experiment it is hard to understand the extent to which planar differential growth rate determines fold positions.

In vitro system:

  1. Can the authors trigger differential growth in their monolayer similar to what has been observed in vivo in Tozluoglu et al. and quantify the extent to which fold position and morphology changes?
  2. Can the position of folds be biased by providing asymmetric confinement?
  3. Is the time-scale of folding comparable to what happens in vivo?

References

  1. Osborn, J. W., A model of growth restraints to explain the development and evolution of tooth shapes in mammals, J. Theor. Biol., 2008.
  2. Shyer, A. E., Tallinen, T., Nerurkar, N. L., Wei, Z., Gil, E. S., Kaplan, D. L., Tabin, C. J., and Mahadevan, L., Villification: How the gut gets its villi, Science, 2013.
  3. Savin, T., Kurpios, N. A., Shyer, A. E., Florescu, P., Liang, H., Mahadevan, L., and Tabin, C. J., On the growth and form of the gut, Nature, 2011.
  4. Varner, V. D., Gleghorn, J. P., Miller, E., Radisky, D. C., and Nelson, C. M., Mechanically patterning the embryonic airway epithelium, Proc. Natl. Acad. Sci., 2015.

Tags: buckling, confinement, fly, mechanics

Posted on: 23rd February 2019 , updated on: 25th February 2019

(No Ratings Yet)




  • Author's response

    Melda Tozluoğlu shared about Planar differential growth rates determine the position of folds in complex epithelia

    We are delighted that our preprint was selected for preLights. We thank Sundar and are looking forward to feedback from the community. We think the highlight provides a clear summary of our work, and the significance in our point of view: Planar differential growth is a driver of precise fold positioning, and robust emergent tissue architecture in buckling epithelia. Some very key open questions are also highlighted, and we are happy to add our commentary on these.

    The patterning of Drosophila wing imaginal disc has been subject to investigation over decades. We know the key morphogen gradients, such as Dpp and wingless, set the overall growth patterns and compartmentalisation of the disc. Regarding the fine details of growth patterns as we observe in our measurements, the simple comment is that what determines such fine-tuning of growth is indeed still an open question and a very interesting one. In our approach, we take the observed growth profiles as given, and investigate their downstream influences on tissue architecture.

    The BM clearly has spatial and temporal heterogeneity; the younger discs seem to harbour thin and relatively uniform BM, whereas the BM of older discs (120hr AEL) is non-homogenous in both its thickness and the visible structure of the fibres. Although the intuitive argument is that a thicker BM would be stiffer, a thickness increase resulting from a sparser fibre network, or alteration of the composition of fibres, could result in reduction of BM stiffness. Clarification of these complexities of the BM structure requires extensive further research and indeed could influence the folding process. Our simulations demonstrate these fine details of the BM are not essential in precise positioning of the wing disc folds, as we can realise them with a homogenous BM definition. These complexities could in fact be functional in progression of the folds.

    The strength of any computational model is demonstrated with its ability to provide testable, accurate predictions. With this philosophy in our work, beyond the definition of fold positioning, we predict perturbations to fold structure upon alterations of the growth rates. In the wing disc, global growth perturbations induce too severe deformations with excessive folding, hindering it impossible to judge perturbations on specific folds. The closest to a controllable, systematic growth perturbation we have in the experimental setup is mutations that naturally alter growth rates at specific regions of the tissue, such as the wingless spade mutant we tested in our study. Here, the mutant reduces growth specifically at the wing hinge, and by utilising only this reduction in growth; we can successfully predict the new emergent fold number and structure. We believe our model predictions and their experimental validation on emergent fold morphology upon perturbation of growth demonstrate the role of differential growth in definition of precise fold architecture.

    and

    Aurélien Roux shared about Buckling of epithelium growing under spherical confinement

    a. It seems a priori difficult to modulate cell growth in a spatially controlled manner. However, we could use two epithelial cell types with different growth rates (and labelled differentially so that they can be traced) and co-encapsulate them. These cells will form islets with different growth rates that will merge into a confluent layer, reproducing the heterogeneity of growth rate. We then could correlate folds position and growth rate.

    b. Indeed, if one look carefully at the picture, folding often occurs where the shell is the thinnest. We thus think that folding is coupled to the local stiffness of the surrounding substrate.

    c. In our experiments, folding takes about 5 to 10 hours, which is fully compatible with the times observed in Tozluoglu et al. (8.5 hours in average), but also for other embryonic folding events like gastrulation or neurulation.

    Have your say

    Your email address will not be published. Required fields are marked *

    This site uses Akismet to reduce spam. Learn how your comment data is processed.

    Sign up to customise the site to your preferences and to receive alerts

    Register here

    Also in the biophysics category:

    The autophagic membrane tether ATG2A transfers lipids between membranes

    Shintaro Maeda, Chinatsu Otomo, Takanori Otomo



    Selected by Sandra Malmgren Hill

    Effective concentrations enforced by intrinsically disordered linkers are governed by polymer physics

    Charlotte S. Sørensen, Magnus Kjaergaard



    Selected by Tessa Sinnige

    1

    Spreading of molecular mechanical perturbations on linear filaments

    Zsombor Balassy, Anne-Marie Lauzon, Lennart Hilbert



    Selected by Lars Hubatsch

    Blue light induces neuronal-activity-regulated gene expression in the absence of optogenetic proteins

    Kelsey M. Tyssowski, Jesse M. Gray



    Selected by Zheng-Shan Chong

    HIV-1 Gag specifically restricts PI(4,5)P2 and cholesterol mobility in living cells creating a nanodomain platform for virus assembly

    C. Favard, J. Chojnacki, P. Merida, et al.



    Selected by Amberley Stephens

    Microfluidic protein isolation and sample preparation for high resolution cryo-EM

    Claudio Schmidli, Stefan Albiez, Luca Rima, et al.



    Selected by David Wright

    ENDOSOMAL MEMBRANE TENSION CONTROLS ESCRT-III-DEPENDENT INTRA-LUMENAL VESICLE FORMATION

    Vincent Mercier, Jorge Larios, Guillaume Molinard, et al.



    Selected by Nicola Stevenson

    1

    Microtubules stabilize intercellular contractile force transmission during tissue folding



    Selected by Ivana Viktorinová

    Dynamic Aha1 Co-Chaperone Binding to Human Hsp90

    Javier Oroz, Laura J Blair, Markus Zweckstetter



    Selected by Reid Alderson

    1

    A DNA-based voltmeter for organelles

    Anand Saminathan, John Devany, Kavya S Pillai, et al.



    Selected by Robert Mahen

    1

    Central spindle microtubules are strongly coupled to chromosomes during both anaphase A and anaphase B

    Che-Hang Yu, Stefanie Redemann, Hai-Yin Wu, et al.



    Selected by Federico Pelisch

    1

    Mechanical Stretch Kills Transformed Cancer Cells

    Ajay Tijore, Mingxi Yao, Yu-Hsiu Wang, et al.



    Selected by Joseph Jose Thottacherry

    Super-resolution Molecular Map of Basal Foot Reveals Novel Cilium in Airway Multiciliated Cells

    Quynh Nguyen, Zhen Liu, Rashmi Nanjundappa, et al.



    Selected by Robert Mahen

    Force inference predicts local and tissue-scale stress patterns in epithelia

    Weiyuan Kong, Olivier Loison, Pruthvi Chavadimane Shivakumar, et al.



    Selected by Sundar Naganathan

    Single molecule localization microscopy with autonomous feedback loops for ultrahigh precision

    Simao Coelho, Jongho Baek, Matthew S Graus, et al.



    Selected by Lars Hubatsch

    Phase transition and amyloid formation by a viral protein as an additional molecular mechanism of virus-induced cell toxicity

    Edoardo Salladini, Claire Debarnot, Vincent Delauzun, et al.



    Selected by Tessa Sinnige

    Also in the developmental biology category:

    The visual system of the genetically tractable crustacean Parhyale hawaiensis: diversification of eyes and visual circuits associated with low-resolution vision

    Ana Patricia Ramos, Ola Gustafsson, Nicolas Labert, et al.



    Selected by Alexa Sadier

    Distinct RhoGEFs activate apical and junctional actomyosin contractility under control of G proteins during epithelial morphogenesis

    Alain Garcia De Las Bayonas, Jean-Marc Philippe, Annemarie C. Lellouch, et al.



    Selected by Ivana Viktorinová

    1

    The Hunchback temporal transcription factor determines motor neuron axon and dendrite targeting in Drosophila

    Austin Q Seroka, Chris Q Doe



    Selected by Abagael Lasseigne

    1

    Preformed Chromatin Topology Assists Transcriptional Robustness of Shh during Limb Development

    Christina Paliou, Philine Guckelberger, Robert Schöpflin, et al.



    Selected by Rafael Galupa

    1

    Molecular Logic of Spinocerebellar Tract Neuron Diversity and Connectivity

    Myungin Baek, Vilas Menon, Thomas Jessell, et al.



    Selected by Yen-Chung Chen

    In vivo glucose imaging in multiple model organisms with an engineered single-wavelength sensor

    Jacob P. Keller, Jonathan S. Marvin, Haluk Lacin, et al.



    Selected by Stephan Daetwyler

    1

    Active behaviour during early development shapes glucocorticoid reactivity

    Luis A. Castillo-Ramírez, Soojin Ryu, Rodrigo J. De Marco



    Selected by Kathleen Gilmour

    A unicellular relative of animals generates an epithelium-like cell layer by actomyosin-dependent cellularization

    Omaya Dudin, Andrej Ondracka, Xavier Grau-Bové, et al.



    Selected by Paul Gerald L. Sanchez and Stefano Vianello

    1

    Members of the Arabidopsis auxin receptor gene family are essential early in embryogenesis and have broadly overlapping functions

    Michael J Prigge, Nikita Kadakia, Kathleen Greenham, et al.



    Selected by Chandra Shekhar Misra

    1

    The spindle assembly checkpoint functions during early development in non-chordate embryos

    Janet Chenevert, Marianne Roca, Lydia Besnardeau, et al.



    Selected by Maiko Kitaoka

    Over-activation of BMP signaling in neural crest cells precipitates heart outflow tract septation

    Jean Francois Darrigrand, Mariana Valente, Pauline Martinez, et al.



    Selected by Giulia Boezio

    Mutations in the Insulator Protein Suppressor of Hairy Wing Induce Genome Instability

    Shih-Jui Hsu, Emily C. Stow, James R. Simmons, et al.



    Selected by Maiko Kitaoka

    1

    Multi-immersion open-top light-sheet microscope for high-throughput imaging of cleared tissues

    Adam K. Glaser, Nicholas P. Reder, Ye Chen, et al.



    Selected by Tim Fessenden

    1

    Dynamic Erasure of Random X-Chromosome Inactivation during iPSC Reprogramming

    Adrian Janiszewski, Irene Talon, Juan Song, et al.



    Selected by Sergio Menchero

    MicroRNA-mediated control of developmental lymphangiogenesis

    Hyun Min Jung, Ciara Hu, Alexandra M Fister, et al.



    Selected by Rudra Nayan Das

    Endogenous CRISPR arrays for scalable whole organism lineage tracing

    James Cotterell, James Sharpe



    Selected by Irepan Salvador-Martinez
    Close