Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Mechanosensing is critical for axon growth in the developing brain

This article has been updated

Abstract

During nervous system development, neurons extend axons along well-defined pathways. The current understanding of axon pathfinding is based mainly on chemical signaling. However, growing neurons interact not only chemically but also mechanically with their environment. Here we identify mechanical signals as important regulators of axon pathfinding. In vitro, substrate stiffness determined growth patterns of Xenopus retinal ganglion cell axons. In vivo atomic force microscopy revealed a noticeable pattern of stiffness gradients in the embryonic brain. Retinal ganglion cell axons grew toward softer tissue, which was reproduced in vitro in the absence of chemical gradients. To test the importance of mechanical signals for axon growth in vivo, we altered brain stiffness, blocked mechanotransduction pharmacologically and knocked down the mechanosensitive ion channel piezo1. All treatments resulted in aberrant axonal growth and pathfinding errors, suggesting that local tissue stiffness, read out by mechanosensitive ion channels, is critically involved in instructing neuronal growth in vivo.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Mechanosensitivity of RGC axons in vitro.
Figure 2: In vivo brain mechanics.
Figure 3: Neurons grow toward soft tissue.
Figure 4: Perturbing brain stiffness leads to axonal pathfinding errors.
Figure 5: In vivo manipulation of mechanosensitive ion channels disrupts axon pathfinding.
Figure 6: Schematics of the mechanical control of axon growth.

Similar content being viewed by others

Change history

  • 03 October 2016

    In the version of this article initially published online, the equation in the last sentence of Methods section "In vitro time-lapse experiments," immediately following "When the third component of ...", contained a dot product. This should have been a cross product. The error has been corrected for the print, PDF and HTML versions of this article.

References

  1. Sperry, R.W. Chemoaffinity in the orderly growth of nerve fiber patterns and connections. Proc. Natl. Acad. Sci. USA 50, 703–710 (1963).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Tessier-Lavigne, M. & Goodman, C.S. The molecular biology of axon guidance. Science 274, 1123–1133 (1996).

    Article  CAS  PubMed  Google Scholar 

  3. Erskine, L. & Herrera, E. The retinal ganglion cell axon's journey: insights into molecular mechanisms of axon guidance. Dev. Biol. 308, 1–14 (2007).

    Article  CAS  PubMed  Google Scholar 

  4. Campbell, D.S. et al. Semaphorin 3A elicits stage-dependent collapse, turning, and branching in Xenopus retinal growth cones. J. Neurosci. 21, 8538–8547 (2001).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Piper, M. et al. Signaling mechanisms underlying Slit2-induced collapse of Xenopus retinal growth cones. Neuron 49, 215–228 (2006).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Atkinson-Leadbeater, K. et al. Dynamic expression of axon guidance cues required for optic tract development is controlled by fibroblast growth factor signaling. J. Neurosci. 30, 685–693 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Chan, C.E. & Odde, D.J. Traction dynamics of filopodia on compliant substrates. Science 322, 1687–1691 (2008).

    Article  CAS  PubMed  Google Scholar 

  8. Moore, S.W., Biais, N. & Sheetz, M.P. Traction on immobilized netrin-1 is sufficient to reorient axons. Science 325, 166 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Betz, T., Koch, D., Lu, Y.B., Franze, K. & Käs, J.A. Growth cones as soft and weak force generators. Proc. Natl. Acad. Sci. USA 108, 13420–13425 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Koch, D., Rosoff, W.J., Jiang, J., Geller, H.M. & Urbach, J.S. Strength in the periphery: growth cone biomechanics and substrate rigidity response in peripheral and central nervous system neurons. Biophys. J. 102, 452–460 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Weiss, P. In vitro experiments on the factors determining the course of the outgrowing nerve fiber. J. Exp. Zool. 68, 393–448 (1934).

    Article  Google Scholar 

  12. Flanagan, L.A., Ju, Y.E., Marg, B., Osterfield, M. & Janmey, P.A. Neurite branching on deformable substrates. Neuroreport 13, 2411–2415 (2002).

    Article  PubMed  PubMed Central  Google Scholar 

  13. Georges, P.C., Miller, W.J., Meaney, D.F., Sawyer, E.S. & Janmey, P.A. Matrices with compliance comparable to that of brain tissue select neuronal over glial growth in mixed cortical cultures. Biophys. J. 90, 3012–3018 (2006).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Kostic, A., Sap, J. & Sheetz, M.P. RPTPalpha is required for rigidity-dependent inhibition of extension and differentiation of hippocampal neurons. J. Cell Sci. 120, 3895–3904 (2007).

    Article  CAS  PubMed  Google Scholar 

  15. Jiang, F.X., Yurke, B., Schloss, R.S., Firestein, B.L. & Langrana, N.A. Effect of dynamic stiffness of the substrates on neurite outgrowth by using a DNA-crosslinked hydrogel. Tissue Eng. Part A 16, 1873–1889 (2010).

    Article  CAS  PubMed  Google Scholar 

  16. Balgude, A.P., Yu, X., Szymanski, A. & Bellamkonda, R.V. Agarose gel stiffness determines rate of DRG neurite extension in 3D cultures. Biomaterials 22, 1077–1084 (2001).

    Article  CAS  PubMed  Google Scholar 

  17. Elkin, B.S., Azeloglu, E.U., Costa, K.D. & Morrison, B. III. Mechanical heterogeneity of the rat hippocampus measured by atomic force microscope indentation. J. Neurotrauma 24, 812–822 (2007).

    Article  PubMed  Google Scholar 

  18. Christ, A.F. et al. Mechanical difference between white and gray matter in the rat cerebellum measured by scanning force microscopy. J. Biomech. 43, 2986–2992 (2010).

    Article  PubMed  Google Scholar 

  19. Franze, K. et al. Spatial mapping of the mechanical properties of the living retina using scanning force microscopy. Soft Matter 7, 3147–3154 (2011).

    Article  CAS  Google Scholar 

  20. Iwashita, M., Kataoka, N., Toida, K. & Kosodo, Y. Systematic profiling of spatiotemporal tissue and cellular stiffness in the developing brain. Development 141, 3793–3798 (2014).

    Article  CAS  PubMed  Google Scholar 

  21. Elkin, B.S., Ilankovan, A. & Morrison, B. III. Age-dependent regional mechanical properties of the rat hippocampus and cortex. J. Biomech. Eng. 132, 011010 (2010).

    Article  PubMed  Google Scholar 

  22. Koser, D.E., Moeendarbary, E., Hanne, J., Kuerten, S. & Franze, K. CNS cell distribution and axon orientation determine local spinal cord mechanical properties. Biophys. J. 108, 2137–2147 (2015).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Weickenmeier, J. et al. Brain stiffness increases with myelin content. Acta Biomater. 42, 265–272 (2016).

    Article  CAS  PubMed  Google Scholar 

  24. Moshayedi, P. et al. Mechanosensitivity of astrocytes on optimized polyacrylamide gels analyzed by quantitative morphometry. J. Phys. Condens. Matter 22, 194114 (2010).

    Article  PubMed  CAS  Google Scholar 

  25. Franze, K., Janmey, P.A. & Guck, J. Mechanics in neuronal development and repair. Annu. Rev. Biomed. Eng. 15, 227–251 (2013).

    Article  CAS  PubMed  Google Scholar 

  26. Guan, W., Puthenveedu, M.A. & Condic, M.L. Sensory neuron subtypes have unique substratum preference and receptor expression before target innervation. J. Neurosci. 23, 1781–1791 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Franze, K. et al. Neurite branch retraction is caused by a threshold-dependent mechanical impact. Biophys. J. 97, 1883–1890 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Kerstein, P.C. et al. Mechanosensitive TRPC1 channels promote calpain proteolysis of talin to regulate spinal axon outgrowth. J. Neurosci. 33, 273–285 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Zhang, Q.Y. et al. Stiff substrates enhance cultured neuronal network activity. Sci. Rep. 4, 6215 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Shibasaki, K., Murayama, N., Ono, K., Ishizaki, Y. & Tominaga, M. TRPV2 enhances axon outgrowth through its activation by membrane stretch in developing sensory and motor neurons. J. Neurosci. 30, 4601–4612 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Pathak, M.M. et al. Stretch-activated ion channel Piezo1 directs lineage choice in human neural stem cells. Proc. Natl. Acad. Sci. USA 111, 16148–16153 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Coste, B. et al. Piezo1 and Piezo2 are essential components of distinct mechanically activated cation channels. Science 330, 55–60 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Coste, B. et al. Piezo proteins are pore-forming subunits of mechanically activated channels. Nature 483, 176–181 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Kim, S.E., Coste, B., Chadha, A., Cook, B. & Patapoutian, A. The role of Drosophila Piezo in mechanical nociception. Nature 483, 209–212 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Li, J. et al. Piezo1 integration of vascular architecture with physiological force. Nature 515, 279–282 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Gottlieb, P.A. & Sachs, F. Piezo1: properties of a cation selective mechanical channel. Channels (Austin) 6, 214–219 (2012).

    Article  CAS  Google Scholar 

  37. Kanekar, S. et al. Xath5 participates in a network of bHLH genes in the developing Xenopus retina. Neuron. 19, 981–994 (1997).

    Article  CAS  PubMed  Google Scholar 

  38. Swift, J. et al. Nuclear lamin-A scales with tissue stiffness and enhances matrix-directed differentiation. Science 341, 1240104 (2013).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  39. Majkut, S. et al. Heart-specific stiffening in early embryos parallels matrix and myosin expression to optimize beating. Curr. Biol. 23, 2434–2439 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Walz, A., Anderson, R.B., Irie, A., Chien, C.B. & Holt, C.E. Chondroitin sulfate disrupts axon pathfinding in the optic tract and alters growth cone dynamics. J. Neurobiol. 53, 330–342 (2002).

    Article  CAS  PubMed  Google Scholar 

  41. Pogoda, K. et al. Compression stiffening of brain and its effect on mechanosensing by glioma cells. New J. Phys. 16, 075002 (2014).

    Article  PubMed  PubMed Central  Google Scholar 

  42. Lo, C.M., Wang, H.B., Dembo, M. & Wang, Y.L. Cell movement is guided by the rigidity of the substrate. Biophys. J. 79, 144–152 (2000).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Isenberg, B.C., Dimilla, P.A., Walker, M., Kim, S. & Wong, J.Y. Vascular smooth muscle cell durotaxis depends on substrate stiffness gradient strength. Biophys. J. 97, 1313–1322 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Bollmann et al. Microglia mechanics: immune activation alters traction forces and durotaxis. Front. Cell Neurosci. 9, 363 (2015).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  45. Xu, G. et al. Opening angles and material properties of the early embryonic chick brain. J. Biomech. Eng. 132, 011005 (2010).

    Article  PubMed  Google Scholar 

  46. Franze, K. The mechanical control of nervous system development. Development 140, 3069–3077 (2013).

    Article  CAS  PubMed  Google Scholar 

  47. Singh, T., Meena, R. & Kumar, A. Effect of sodium sulfate on the gelling behavior of agarose and water structure inside gel networks. J. Phys. Chem. B 113, 2519–2525 (2009).

    Article  CAS  PubMed  Google Scholar 

  48. Höpker, V.H., Shewan, D., Tessier-Lavigne, M., Poo, M. & Holt, C. Growth-cone attraction to netrin-1 is converted to repulsion by laminin-1. Nature 401, 69–73 (1999).

    Article  PubMed  Google Scholar 

  49. Nieuwkoop, P.D. & Faber, J. Normal table of Xenopus laevis (Daudin): a Systematical and Chronological Survey of the Development from the Fertilized Egg till the End of Metamorphosis (North-Holland Pub. Co., 1967).

  50. Das, T., Payer, B., Cayouette, M. & Harris, W.A. In vivo time-lapse imaging of cell divisions during neurogenesis in the developing zebrafish retina. Neuron 37, 597–609 (2003).

    Article  CAS  PubMed  Google Scholar 

  51. Chien, C.B., Rosenthal, D.E., Harris, W.A. & Holt, C.E. Navigational errors made by growth cones without filopodia in the embryonic Xenopus brain. Neuron 11, 237–251 (1993).

    Article  CAS  PubMed  Google Scholar 

  52. Leung, K.M. et al. Asymmetrical beta-actin mRNA translation in growth cones mediates attractive turning to netrin-1. Nat. Neurosci. 9, 1247–1256 (2009).

    Article  CAS  Google Scholar 

  53. Kalous, A., Stake, J.I., Yisraeli, J.K. & Holt, C.E. RNA-binding protein Vg1RBP regulates terminal arbor formation but not long-range axon navigation in the developing visual system. Dev. Neurobiol. 74, 303–318 (2014).

    Article  CAS  PubMed  Google Scholar 

  54. Wizenmann, A. et al. Extracellular Engrailed participates in the topographic guidance of retinal axons in vivo. Neuron 64, 355–366 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  55. Hutter, J.L. & Bechhoefer, J. Calibration of atomic-force microscope tips. Rev. Sci. Instrum. 64, 1868–1873 (1993).

    Article  CAS  Google Scholar 

  56. Mahaffy, R.E., Shih, C.K., MacKintosh, F.C. & Käs, J. Scanning probe-based frequency-dependent microrheology of polymer gels and biological cells. Phys. Rev. Lett. 85, 880–883 (2000).

    Article  CAS  PubMed  Google Scholar 

  57. Franze, K. Atomic force microscopy and its contribution to understanding the development of the nervous system. Curr. Opin. Genet. Dev. 21, 530–537 (2011).

    Article  CAS  PubMed  Google Scholar 

  58. Sholl, D.A. Dendritic organization in the neurons of the visual and motor cortices of the cat. J. Anat. 87, 387–406 (1953).

    CAS  PubMed  PubMed Central  Google Scholar 

  59. Hertz, H. Über die Berührung fester elastischer Körper. J. Reine Angew. Math. 92, 156–171 (1881).

    Google Scholar 

  60. Tamada, A., Kawase, S., Murakami, F. & Kamiguchi, H. Autonomous right-screw rotation of growth cone filopodia drives neurite turning. J. Cell Biol. 188, 429–441 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We thank D. Bray, A. Reichenbach, B. Simons and S. Wolff for discussions about neuronal mechanics, K. Chalut and W. Harris for feedback on the manuscript, A. Christ for an AFM data analysis code, P. Moshayedi for help with establishing PAA gels, F. Sachs (University at Buffalo, USA) for providing GsMTx4, H. Wong for extensive help in the lab and A. Winkel and R. Field (JPK) for technical help. This work was supported by the German National Academic Foundation (scholarship to D.E.K.), Wellcome Trust and Cambridge Trusts (scholarships to A.J.T.), Winston Churchill Foundation of the United States (scholarship to S.K.F.), Herchel Smith Foundation (Research Studentship to S.K.F.), CNPq 307333/2013-2 (L.d.F.C.), NAP-PRP-USP and FAPESP 11/50761-2 (L.d.F.C.), UK EPSRC BT grant (J.G.), Wellcome Trust WT085314 and the European Research Council 322817 grants (C.E.H.); an Alexander von Humboldt Foundation Feodor Lynen Fellowship (K.F.), UK BBSRC grant BB/M021394/1 (K.F.), the Human Frontier Science Program Young Investigator Grant RGY0074/2013 (K.F.), the UK Medical Research Council Career Development Award G1100312/1 (K.F.) and the Eunice Kennedy Shriver National Institute Of Child Health & Human Development of the National Institutes of Health under Award Number R21HD080585 (K.F.). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

Author information

Authors and Affiliations

Authors

Contributions

K.F. conceived the project; J.G., C.E.H. and K.F. designed the research; D.E.K., A.J.T., S.K.F., A.D., G.K.S., E.K.P., H.S. and K.F. performed the experiments; D.E.K., A.J.T., S.K.F., G.K.S., E.K.P., H.S., M.V., L.d.F.C. and K.F. analyzed the data; all authors discussed the data; K.F. wrote the manuscript with contributions from all authors.

Corresponding author

Correspondence to Kristian Franze.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Integrated supplementary information

Supplementary Figure 1 In vitro stiffness gradient culture substrates.

(a) Description of chamber assembly. Premixes of stiff and soft gels were prepared (with fluorescently tagged stiff polymer), polymerization was initiated, polymerizing stiff gel filled into the chamber (to fill half of it), which was then overlaid with polymerizing soft gel. At the interface between the two solutions, polymerizing gels mixed due to diffusion, resulting in a gradient in stiffness and fluorescence signal in the polymerized gel. (b) AFM stiffness map of a gel stiffness gradient. (c) Laminin coating of PAA gels. Immunofluorescence showed equal coating densities on soft and stiff gels, indicating homogeneous coating of the substrate surface (Mann-Whitney-Test; P = 0.430, Z = 0.789). n = number of randomly selected regions of interest from three independent experiments. (d) Quantification of stiffness gradients. Stiffness gradients were sigmoidal, with a large (~3 mm) linear region (quality of a linear fit to the linear region: R2 = 0.99 ± 0.01 (mean ± S.E.M.), n = 8). The average slope of the stiffness gradients was Mmax = (1.8 ± 0.3) Pa/µm (n = 8). (e) There was a strong correlation between local substrate stiffness and fluorescence intensity (Pearson’s correlation coefficient r > 0.98 for all measured substrates (n = 4)), enabling the use of fluorescence signal as readout for K.

Supplementary Figure 2 Quantification of axon growth on compliant culture substrates.

(a-d) Axon growth on compliant substrates coated with fibronectin. (a, b) Cultures of Xenopus eye primordia on (a) ‘soft’ (0.1 kPa) and (b) ‘stiff’ (1 kPa) substrates. Shown are representative images from three independent experiments. (c) Sholl analysis of axon lengths after 24 hours (normalized counts as mean ± S.E.M.). (d) Box plots of the median distances shown in (c). Axons were significantly longer on stiffer substrates than on soft ones (two-tailed t-test; P = 1.82 × 10-6, t = 6.118). Although fibronectin engages different integrins than laminin, RGC axons assumed similar phenotypes on soft and stiff substrates in these experiments, indicating that neurons were mechanosensitive irrespective of the type of integrins involved in cell adhesion. n = number of eye primordia from three independent experiments. (e-g) Directionality of axon growth on compliant substrates coated with laminin. RGC axon growth directions are more random on soft (e) than on stiff (f) substrates. Rose diagrams show the distribution of turning angles of growth cones. 0° corresponds to a straight forward movement, 90° to a left-turn, and 180° corresponds to retractions. Each ring in the rose plot represents the labelled percentage of the total number of angles. The proportion of axons growing straight forward is significantly larger on stiffer substrates than on softer ones (for axons growing between 355° and 5°: P = 3.88 x 10-4, z = 3.548, Mann-Whitney-test, n = 60 each for soft and stiff). (g) Quantification of Fig. 1j. A significantly larger percentage of axons was deviating by more than 20° from a straight path on soft compared to stiff substrates (two-tailed t-test; P = 0.026, t = 2.274), showing that axons grew less ordered on softer substrates. n = number of randomly selected regions of interest from three independent experiments.

Supplementary Figure 3 Elasticity maps for a fixed indentation depth.

(a, b, d) Same samples as shown in Figs. 2c, d, and 4a, respectively; here K was determined not at a defined force but at a defined indentation depth δ = 3 µm. Stiffness gradients in the tissue are very similar to those shown in Figs. 2 and 4. Blue lines indicate outlines of the OT. Insets in (b) and (d): epifluorescence images of RGC axons in the OT of the respective brains. (c) Stiffness map of a brain with ablated RGC axons. Similar stiffness gradients are found as in control brains, indicating that gradients likely arise from the distribution of cell somata in the tissue (cf. Fig. 2e). Scale bar: 200 µm. Experiments were repeated three times and representative images are shown.

Supplementary Figure 4 In vivo stiffness gradients.

(a) Relationship between local gradients in tissue stiffness perpendicular to the RGC axon growth direction, M, and local OT curvature, C, in Xenopus brains treated with 15 mg/ml CS. (b) Same data as in (a), pooled (green). Blue box plot (controls) replotted from Fig. 3c for comparison. Also in CS-treated brains, axons in vivo preferentially turned towards the softer side of the tissue (one-sample Wilcoxon Signed Rank Test; P = 5.63 × 10-4, Z = 3.449). There was no statistical difference between control and CS-treated animals (Mann-Whitney-Test; P = 0.354, Z = 0.927). n = number of measurement points from nine CS-treated animals. (c, d) Relationship between the absolute stiffness K of the gradient RGC axons encountered in vivo and curvature C of the OT. We did not find a correlation between K and curvature in either control (c) or CS-treated (d) brains; only the gradient strength M is important (cf. Fig. 3).

Supplementary Figure 5 Local mechanical manipulation of brain tissue redirects axon growth.

(a) Image of an AFM experiment. A sustained stress was applied with a cantilever, likely leading to a stiffening of the tissue underneath the spherical probe (outline indicated by black circle). Axons (surrounded by dashed curve, arrow) had started growing along the brain surface but had not reached the mid-diencephalon yet. Scale bar: 200 µm. (b,c,f,g) Images showing outlines of Xenopus brains and the OT (outlined by dashed curve) at the end of four independent experiments. The position of the AFM probe is shown by a filled circle. Axons avoided the tissue underneath the bead and in some cases (b, c) grew around the stiffened tissue. (d) and (e) show magnified regions indicated by dashed rectangles in (b) and (c), respectively. Dashed circles indicate AFM probe positions. Scale bars: 100 µm. (h) Compression led to significant tissue stiffening (n = 13 from four different animals, Kruskal-Wallis ANOVA; P = 0.00683, χ2 = 9.97396).

Supplementary Figure 6 Piezo1 in Xenopus RGC axons.

(a-c) MO knockdown of Piezo1 led to aberrant axon growth in vivo, which was maintained at stage 42 (~1 day after the experiments shown in Fig. 5). Scale bar: 100 µm. Representative images from three independent experiments. (c) Quantification of OT morphology. At stage 42, elongation was still significantly decreased (two-tailed t-test; P = 5.21 × 10-4; t = 3.902), showing that the knockdown did not merely slow down axon growth but rather led to a ‘soft phenotype’ with shorter RGC axons that deviated from their normal path. n = number of animals from three experimental days. (d, e) Western blot analysis confirmed that Piezo1 morpholinos led to a significant decrease in the expression of the protein by 42 % ± 9 % if compared to the scrambled control (n = 4 Western blots from 2 lysates; One-sample t-test; P = 0.00689, t = 6.634). (d) Cropped lanes of Piezo1 and a-tubulin. (e) Full-length blot with molecular weight standard.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–6 (PDF 967 kb)

Supplementary Methods Checklist (PDF 435 kb)

Time-lapse movie of Xenopus retinal ganglion cell axons cultured on a soft substrate of G' = 100 Pa.

1 s corresponds to 3.5 min of real time. Scale bar = 25 μm. (AVI 3960 kb)

Time-lapse movie of Xenopus retinal ganglion cell axons cultured on a soft substrate of G' = 1,000 Pa.

1 s corresponds to 3.5 min of real time. Scale bar = 25 μm. (AVI 1811 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Koser, D., Thompson, A., Foster, S. et al. Mechanosensing is critical for axon growth in the developing brain. Nat Neurosci 19, 1592–1598 (2016). https://doi.org/10.1038/nn.4394

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nn.4394

This article is cited by

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing