Measuring cell adhesion forces of primary gastrulating cells from zebrafish using atomic force microscopy

P. Puech, A. Taubenberger, F. Ulrich, M. Krieg, D. Mueller, C.-P.J. Heisenberg, Journal of Cell Science 118 (2005) 4199–4206.

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Journal Article | Published | English
Author
Puech, Pierre; Taubenberger, Anna; Ulrich, Florian; Krieg, Michael; Mueller, Daniel; Heisenberg, Carl-Philipp IST Austria
Abstract
During vertebrate gastrulation, progenitor cells of different germ layers acquire specific adhesive properties that contribute to germ layer formation and separation. Wnt signals have been suggested to function in this process by modulating the different levels of adhesion between the germ layers, however, direct evidence for this is still lacking. Here we show that Wnt11, a key signal regulating gastrulation movements, is needed for the adhesion of zebrafish mesendodermal progenitor cells to fibronectin, an abundant extracellular matrix component during gastrulation. To measure this effect, we developed an assay to quantify the adhesion of single zebrafish primary mesendodermal progenitors using atomic-force microscopy (AFM). We observed significant differences in detachment force and work between cultured mesendodermal progenitors from wild-type embryos and from slb/wit11 mutant embryos, which carry a loss-of-function mutation in the wnt11 gene, when tested on fibronectin-coated substrates. These differences were probably due to reduced adhesion to the fibronectin substrate as neither the overall cell morphology nor the cell elasticity grossly differed between wild-type and mutant cells. Furthermore, in the presence of inhibitors of fibronectin-integrin binding, such as RGD peptides, the adhesion force and work were strongly decreased, indicating that integrins are involved in the binding of mesendodermal progenitors in our assay. These findings demonstrate that AFM can be used to quantitatively determine the substrate-adhesion of cultured primary gastrulating cells and provide insight into the role of Wnt11 signalling in modulating cell adhesion at the single cell scale.
Publishing Year
Date Published
2005-01-01
Journal Title
Journal of Cell Science
Volume
118
Issue
18
Page
4199 - 4206
IST-REx-ID

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Puech P, Taubenberger A, Ulrich F, Krieg M, Mueller D, Heisenberg C-PJ. Measuring cell adhesion forces of primary gastrulating cells from zebrafish using atomic force microscopy. Journal of Cell Science. 2005;118(18):4199-4206. doi:10.1242/​jcs.02547
Puech, P., Taubenberger, A., Ulrich, F., Krieg, M., Mueller, D., & Heisenberg, C.-P. J. (2005). Measuring cell adhesion forces of primary gastrulating cells from zebrafish using atomic force microscopy. Journal of Cell Science, 118(18), 4199–4206. https://doi.org/10.1242/​jcs.02547
Puech, Pierre, Anna Taubenberger, Florian Ulrich, Michael Krieg, Daniel Mueller, and Carl-Philipp J Heisenberg. “Measuring Cell Adhesion Forces of Primary Gastrulating Cells from Zebrafish Using Atomic Force Microscopy.” Journal of Cell Science 118, no. 18 (2005): 4199–4206. https://doi.org/10.1242/​jcs.02547.
P. Puech, A. Taubenberger, F. Ulrich, M. Krieg, D. Mueller, and C.-P. J. Heisenberg, “Measuring cell adhesion forces of primary gastrulating cells from zebrafish using atomic force microscopy,” Journal of Cell Science, vol. 118, no. 18, pp. 4199–4206, 2005.
Puech P, Taubenberger A, Ulrich F, Krieg M, Mueller D, Heisenberg C-PJ. 2005. Measuring cell adhesion forces of primary gastrulating cells from zebrafish using atomic force microscopy. Journal of Cell Science. 118(18), 4199–4206.
Puech, Pierre, et al. “Measuring Cell Adhesion Forces of Primary Gastrulating Cells from Zebrafish Using Atomic Force Microscopy.” Journal of Cell Science, vol. 118, no. 18, Company of Biologists, 2005, pp. 4199–206, doi:10.1242/​jcs.02547.

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