Quantitative Analysis of Melanocyte Migration in vitro Based on Automated Cell Tracking under Phase Contrast Microscopy Considering the Combined Influence of Cell Division and Cell-Matrix Interactions

V. Letort; S. Fouliard; G. Letort; I. Adanja; M. Kumasaka; S. Gallagher; O. Debeir; L. Larue; F. Xavier

Mathematical Modelling of Natural Phenomena (2010)

  • Volume: 5, Issue: 1, page 4-33
  • ISSN: 0973-5348

Abstract

top
The aim of this study was to describe and analyze the regulation and spatio-temporal dynamics of melanocyte migration in vitro and its coupling to cell division and interaction with the matrix. The melanocyte lineage is particularly interesting because it is involved in both embryonic development and oncogenesis/metastasis (melanoma). Biological experiments were performed on two melanocyte cell lines established from wild-type and β-catenin-transgenic mice (bcat*). The multi-functional β-catenin molecule is known to be able to regulate the transcription of various genes involved in cell proliferation and migration, particularly in the melanocyte lineage. We also investigated fibronectin, an extra-cellular matrix protein that binds integrins, thereby providing adhesion points for cells and encouraging migration. As the migration of individual cells were followed, automated methods were required for processing the large amount of data generated by the time-lapse video-microscopy. A model-based approach for automated cell tracking was evaluated on a sample by comparison with manual tracking. This method was found reliable in studying overall cell behaviour. Its application to all the observed sequences provided insight into the factors affecting melanocyte migration in vitro: melanocytes of mutated form of β-catenin showed higher division rates and no contact inhibition of growth was induced by the resulting increase in cell density. However, cell density limited the amplitude of cell displacements, although their motility was less affected. The high fibronectin concentration bound to substratum promoted cell migration and motility, the effect being more intense for wild-type cells than for cells with β-catenin over-expression. During the division process, cell migration speed increased rapidly then decreased slowly. Analyses of such data is expected to lead both to biological answers and to a framework for a better modeling processes in the future.

How to cite

top

Letort, V., et al. "Quantitative Analysis of Melanocyte Migration in vitro Based on Automated Cell Tracking under Phase Contrast Microscopy Considering the Combined Influence of Cell Division and Cell-Matrix Interactions." Mathematical Modelling of Natural Phenomena 5.1 (2010): 4-33. <http://eudml.org/doc/197685>.

@article{Letort2010,
abstract = {The aim of this study was to describe and analyze the regulation and spatio-temporal dynamics of melanocyte migration in vitro and its coupling to cell division and interaction with the matrix. The melanocyte lineage is particularly interesting because it is involved in both embryonic development and oncogenesis/metastasis (melanoma). Biological experiments were performed on two melanocyte cell lines established from wild-type and β-catenin-transgenic mice (bcat*). The multi-functional β-catenin molecule is known to be able to regulate the transcription of various genes involved in cell proliferation and migration, particularly in the melanocyte lineage. We also investigated fibronectin, an extra-cellular matrix protein that binds integrins, thereby providing adhesion points for cells and encouraging migration. As the migration of individual cells were followed, automated methods were required for processing the large amount of data generated by the time-lapse video-microscopy. A model-based approach for automated cell tracking was evaluated on a sample by comparison with manual tracking. This method was found reliable in studying overall cell behaviour. Its application to all the observed sequences provided insight into the factors affecting melanocyte migration in vitro: melanocytes of mutated form of β-catenin showed higher division rates and no contact inhibition of growth was induced by the resulting increase in cell density. However, cell density limited the amplitude of cell displacements, although their motility was less affected. The high fibronectin concentration bound to substratum promoted cell migration and motility, the effect being more intense for wild-type cells than for cells with β-catenin over-expression. During the division process, cell migration speed increased rapidly then decreased slowly. Analyses of such data is expected to lead both to biological answers and to a framework for a better modeling processes in the future.},
author = {Letort, V., Fouliard, S., Letort, G., Adanja, I., Kumasaka, M., Gallagher, S., Debeir, O., Larue, L., Xavier, F.},
journal = {Mathematical Modelling of Natural Phenomena},
keywords = {melanocytes; automated cell tracking; video microscopy; cell migration; cell division; cell-matrix interaction; β-catenin; fibronectin.; -catenin; fibronectin},
language = {eng},
month = {2},
number = {1},
pages = {4-33},
publisher = {EDP Sciences},
title = {Quantitative Analysis of Melanocyte Migration in vitro Based on Automated Cell Tracking under Phase Contrast Microscopy Considering the Combined Influence of Cell Division and Cell-Matrix Interactions},
url = {http://eudml.org/doc/197685},
volume = {5},
year = {2010},
}

TY - JOUR
AU - Letort, V.
AU - Fouliard, S.
AU - Letort, G.
AU - Adanja, I.
AU - Kumasaka, M.
AU - Gallagher, S.
AU - Debeir, O.
AU - Larue, L.
AU - Xavier, F.
TI - Quantitative Analysis of Melanocyte Migration in vitro Based on Automated Cell Tracking under Phase Contrast Microscopy Considering the Combined Influence of Cell Division and Cell-Matrix Interactions
JO - Mathematical Modelling of Natural Phenomena
DA - 2010/2//
PB - EDP Sciences
VL - 5
IS - 1
SP - 4
EP - 33
AB - The aim of this study was to describe and analyze the regulation and spatio-temporal dynamics of melanocyte migration in vitro and its coupling to cell division and interaction with the matrix. The melanocyte lineage is particularly interesting because it is involved in both embryonic development and oncogenesis/metastasis (melanoma). Biological experiments were performed on two melanocyte cell lines established from wild-type and β-catenin-transgenic mice (bcat*). The multi-functional β-catenin molecule is known to be able to regulate the transcription of various genes involved in cell proliferation and migration, particularly in the melanocyte lineage. We also investigated fibronectin, an extra-cellular matrix protein that binds integrins, thereby providing adhesion points for cells and encouraging migration. As the migration of individual cells were followed, automated methods were required for processing the large amount of data generated by the time-lapse video-microscopy. A model-based approach for automated cell tracking was evaluated on a sample by comparison with manual tracking. This method was found reliable in studying overall cell behaviour. Its application to all the observed sequences provided insight into the factors affecting melanocyte migration in vitro: melanocytes of mutated form of β-catenin showed higher division rates and no contact inhibition of growth was induced by the resulting increase in cell density. However, cell density limited the amplitude of cell displacements, although their motility was less affected. The high fibronectin concentration bound to substratum promoted cell migration and motility, the effect being more intense for wild-type cells than for cells with β-catenin over-expression. During the division process, cell migration speed increased rapidly then decreased slowly. Analyses of such data is expected to lead both to biological answers and to a framework for a better modeling processes in the future.
LA - eng
KW - melanocytes; automated cell tracking; video microscopy; cell migration; cell division; cell-matrix interaction; β-catenin; fibronectin.; -catenin; fibronectin
UR - http://eudml.org/doc/197685
ER -

References

top
  1. M. Abal, M. Piel, V. Bouckson-Castaing, M. Mogensen, J.-B. Sibarita M. Bornens. Microtubule release from the centrosome in migrating cells . The Journal of Cell Biology, 159 (2002), 731–737 
  2. S. Allard, P. Adin, L. Goudard, N. di Clemente, N. Josso, M.-C. Orgebin-Crist, J.-Y. Picard F. Xavier. Molecular mechanisms of hormone-mediated Müllerian duct regression: Involment of beta-catenin. Development, 127 (2000), 3349–360 
  3. K.J. Busam, C. Charles, G. Lee A.C. Halpern. Morphologic Features of Melanocytes, Pigmented Keratinocytes, and Melanophages by In Vivo Confocal Scanning Laser Microscopy. Mod. Pathol., 14 (2001), 862–868 
  4. Y. Cheng. Mean shift, mode seeking, and clustering. IEEE Trans. Pattern Anal. Mach. Intell., 17 (1995), No. 8, 790–799 
  5. A. Dammermann, A. Desai K. Oegema. The minus end in sight. Current Biology, 13 (2003), 614–624 
  6. O. Debeir, I. Camby, R. Kiss, P. Van Ham C. Decaescker. A Model-Based Approach for Automated In Vitro Cell Tracking and Chemotaxis Analyses. Cytometry, 60A (2004), 29–40 
  7. O. Debeir, P. Van Ham, R. Kiss C. Decaestecker. Tracking of Migrating Cells under Phase-contrast Video Microscopy with Combined Mean-Shift Processes. IEEE Transaction on Medical Imaging., 24 (2005), No. 6, 697–711 
  8. O. Debeir, V. Mégalizzi, N. Warzée, R. Kiss C. Decaestecker. Videomicroscopic extraction of specific information on cell proliferation and migration in vitro. Experim. Cell Res., 314 (2008), 2985–2998 
  9. V. Delmas, F. Beermann, S. Martinozzi, S. Carreira, J. Ackermann, M. Kumasaka, L. Denat, J. Goodall, F. Luciani, A. Viros, N. Demirkan, B.C. Bastian, C.R. Goding L. Larue. beta-catenin induces immortalisation of melanocytes by suppressing p16INK4a expression and co-operates with N-Ras in melanoma development. Genes & Dev., 21 (2007), 2923-2935 
  10. N. Desban, J-L. Duband. Avian neural crest cell migration on laminin: interaction of the a11 integrin with distinct laminin-1 domains mediates different adhesive responses. J. Cell Sci.110 (1997), 2729–2744 
  11. C. Decaestecker, O. Debeir, P. Van Ham R. Kiss. Can anti-migratory drugs be screened in vitro ? A review of 2D and 3D assays for the quantitative analysis of cell migration. Inc. Med. Res. Rev., 27 (2007), No. 2, 149–176 
  12. R Development Core Team. R: A Language and Environment for Statistical Computing. R Foundation for Statistical Computing, Vienna, Austria., (2008),  URIhttp://www.R-project.org
  13. P.A. DiMilla, J.A. Stone, J.A. Quinn, S.M. Albelda D.A. Lauffenburger. Maximal Migration of Human Smooth Muscle Cells on Fibronectin and Type IV Collagen Occurs at an Intermediate Attachment Strength. J Cell Biol., 122 (1993), 729–37 
  14. A.J. H, T. Sun, D.C. Barber, D.R. Hose S. Macneil. Automated tracking of migrating cells in phase-contrast video microscopy sequences using image registration. J. Microscopy., 234 (2009), 62–79 
  15. N. Hamilton. Quantification and its Applications in Fluorescent Microscopy Imaging. Traffic, 10 (2009), 951961 
  16. S. Hazgui, N. Bonnet, J. Cutrona, B. Nawrocki-Raby, M. Polette, L. Chouchane, P. Birembaut J-M. Zahm. 3D culture model and computer assisted videomicroscopy to analyze migratory behavior of noninvasive and invasive bronchial epithelial cells. Am. J. Physiol. Cell Physiol., 289 (2005), 1547–1552 
  17. S.P. Holly, M.K. Larson L.V. Parise. Minireview. Multiple Roles of Integrins in Cell Motility. Exp. Cell Res., 261 (2000), 69-74 
  18. A. Jouneau, Y.-Q. YU, M. Pasdar L. Larue. Plasticity of Cadherin-Catenin Expression in the Melanocyte Lineage. Pigment Cell Res.13 (2000), No. 4, 260–272 
  19. E. Knust W.B. Huttner. Cell polarity from cell division. Dev. Cell., 12 (2007), 664–666 
  20. L. Larue, N. Dougherty, S. Porter B. Mintz. Spontaneous malignant transformation of melanocytes explanted from Wf/Wf mice with a Kit kinase-domain mutation. Proc. Natl. Acad. Sci. USA., 89 (1992), 7816–7820 
  21. L. Larue, M. Kumasaka C.R. Goding. Beta-catenin in the melanocyte lineage. Pigment Cell R., 16 (2003), 312–317 
  22. Y. Lee, L.V. McIntire K. Zygourakis. Analysis of endothelial cell locomotion: Differential effects of motility and contact inhibition. Biotechnol Bioeng.43 (1994), 622–634 
  23. T.C. Mayer. The migratory pathway of neural crest cells into the skin of mouse embryos. Dev. Biol., 34 (1973), 39–46 
  24. J. McCarthy E.A. Turley. Effects of Extracellular Matrix Components on Cell Locomotion. Critical Reviews in Oral Biology and Medecine., 4 (1993), 619–637 
  25. D.J. Mooney, R. Langer D.E. Ingber. Cytoskeletal filament assembly and the control of cell spreading and function by extracellular matrix. J. Cell Sci., 108 (1995), 2311-2320 
  26. S.P. Palecek, J.C. Loftus, M.H. Ginsberg, D.A. Lauffenburger A.F. Horwitz. Integrin-ligand binding properties govern cell migration speed through cell-substratum adhesiveness. Nature, 385 (1997), 537–540 
  27. A.J. Ridley, M.A. Schwartz, K. Burridge, R.A. Firtel, M.H. Ginsberg, G. Borisy, J.T. Parsons A.R. Horwitz. Cell migration: integrating signals from front to back. Science., 302 (2003), 1704–1709 
  28. C. Rosello, P. Ballet, E. Planus P. Tracqui. Model driven quantification of individual and collective cell migration. Acta Biotheoretica, 52 (2004), 343-363 
  29. G. Scott, S. Leopardi, S. Printup B.C. Madden. Filopodia are conduits for melanosome transfer to keratinocytes. J. Cell Sci., 115 (2002), 1441–1451 
  30. A. Stéphanou, E. Mylona, M. Chaplain P. Tracqui. A computational model of cell migration coupling the growth of focal adhesions with oscillatory cell protusions. Journal of Theor. Biol., 253 (2008), 701–716 
  31. M. Tawk, C. Araya, D.A. Lyons, A.M. Reugels, G.C. Girdler, P.R. Bayley, D.R. Hyde, M. Tada J.D. Clarke. A mirror-symmetric cell division that orchestrates neuroepithelial morphogenesis. Nature, 446 (2007), No. 7137, 797-800 
  32. M. Théry, V. Racine, A. Pépin, M. Piel, Y. Chen, J.-B. Sibarita M. Bornens. The extracellular matrix guides the orientation of the cell division axis. Nature Cell Biology, 7 (2005), No. 10, 947–953 
  33. A.L. Wilkie, S.A. Jordan I.J. Jackson. Neural crest progenitors of the melanocyte lineage: coat colour patterns revisited. Development, 129 (2002), 3349-57 

NotesEmbed ?

top

You must be logged in to post comments.

To embed these notes on your page include the following JavaScript code on your page where you want the notes to appear.

Only the controls for the widget will be shown in your chosen language. Notes will be shown in their authored language.

Tells the widget how many notes to show per page. You can cycle through additional notes using the next and previous controls.

    
                

Note: Best practice suggests putting the JavaScript code just before the closing </body> tag.