The Journal of Cell Biology, Vol 120, 935-945, Copyright © 1993 by The Rockefeller University Press

ARTICLES

Observation and quantification of individual microtubule behavior in vivo: microtubule dynamics are cell-type specific

E Shelden and P Wadsworth
Department of Biology, University of Massachusetts, Amherst 01003.

Recent experiments have demonstrated that the behavior of the interphase microtubule array is cell-type specific: microtubules in epithelial cells are less dynamic than microtubules in fibroblasts (Pepper-kok et al., 1990; Wadsworth and McGrail, 1990). To determine which parameters of microtubule dynamic instability behavior are responsible for this difference, we have examined the behavior of individual microtubules in both cell types after injection with rhodamine-labeled tubulin subunits. Individual microtubules in both cell types were observed to grow, shorten, and pause, as expected. The average amount of time microtubules remained within the lamellae of CHO fibroblasts, measured from images acquired at 10-s intervals, was significantly shorter than the average amount of time microtubules remained within lamellae of PtK1 epithelial cells. Further analysis of individual microtubule behavior from images acquired at 2-s intervals reveals that microtubules in PtK1 cells undergo multiple brief episodes of growth and shortening, resulting in little overall change in the microtubule network. In contrast, microtubules in lamellae of CHO fibroblasts are observed to undergo fewer transitions which are of longer average duration, resulting in substantial changes in the microtubule network over time. A small subset of more stable microtubules was also detected in CHO fibroblasts. Quantification of the various parameters of dynamic instability behavior from these sequences demonstrates that the average rates of both growth and shortening are significantly greater for the majority of microtubules in fibroblasts than for microtubules in epithelial cells (19.8 +/- 10.8 microns/min, 32.2 +/- 17.7 microns/min, 11.9 +/- 6.5 microns/min, and 19.7 +/- 8.1 microns/min, respectively). The frequency of catastrophe events (1/interval between catastrophe events) was similar in both cell types, but the frequency of rescue events (1/time spent shrinking) was significantly higher in PtK1 cells. Thus, individual microtubules in PtK1 lamellae undergo frequent excursions of short duration and extent, whereas most microtubules in CHO lamellae undergo more extensive excursions often resulting in the appearance or disappearance of microtubules within the field of view. These observations provide the first direct demonstration of cell-type specific behavior of individual microtubules in living cells, and indicate that these differences can be brought about by modulation of the frequency of rescue. These results directly support the view that microtubule dynamic instability behavior is regulated in a cell-type specific manner.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Facebook    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

• Tanabe, K., Takei, K. (2009). Dynamic instability of microtubules requires dynamin 2 and is impaired in a Charcot-Marie-Tooth mutant. JCB 185: 939-948 [Abstract] [Full Text]  
• Efimov, A., Schiefermeier, N., Grigoriev, I., Brown, M. C., Turner, C. E., Small, J. V., Kaverina, I. (2008). Paxillin-dependent stimulation of microtubule catastrophes at focal adhesion sites. J. Cell Sci. 121: 196-204 [Abstract] [Full Text]  
• Ichii, T., Takeichi, M. (2007). p120-catenin regulates microtubule dynamics and cell migration in a cadherin-independent manner. GENES CELLS 12: 827-839 [Abstract] [Full Text]  
• Efimov, V. P., Zhang, J., Xiang, X. (2006). CLIP-170 Homologue and NUDE Play Overlapping Roles in NUDF Localization in Aspergillus nidulans. Mol. Biol. Cell 17: 2021-2034 [Abstract] [Full Text]  
• Reilein, A., Yamada, S., Nelson, W. J. (2005). Self-organization of an acentrosomal microtubule network at the basal cortex of polarized epithelial cells. JCB 171: 845-855 [Abstract] [Full Text]  
• Salaycik, K. J., Fagerstrom, C. J., Murthy, K., Tulu, U. S., Wadsworth, P. (2005). Quantification of microtubule nucleation, growth and dynamics in wound-edge cells. J. Cell Sci. 118: 4113-4122 [Abstract] [Full Text]  
• Mimori-Kiyosue, Y., Grigoriev, I., Lansbergen, G., Sasaki, H., Matsui, C., Severin, F., Galjart, N., Grosveld, F., Vorobjev, I., Tsukita, S., Akhmanova, A. (2005). CLASP1 and CLASP2 bind to EB1 and regulate microtubule plus-end dynamics at the cell cortex. JCB 168: 141-153 [Abstract] [Full Text]  
• Tirnauer, J. S., Salmon, E. D., Mitchison, T. J. (2004). Microtubule Plus-End Dynamics in Xenopus Egg Extract Spindles. Mol. Biol. Cell 15: 1776-1784 [Abstract] [Full Text]  
• Honore, S., Kamath, K., Braguer, D., Wilson, L., Briand, C., Jordan, M. A. (2003). Suppression of microtubule dynamics by discodermolide by a novel mechanism is associated with mitotic arrest and inhibition of tumor cell proliferation. Molecular Cancer Therapeutics 2: 1303-1311 [Abstract] [Full Text]  
• Shirasu-Hiza, M., Coughlin, P., Mitchison, T. (2003). Identification of XMAP215 as a microtubule-destabilizing factor in Xenopus egg extract by biochemical purification. JCB 161: 349-358 [Abstract] [Full Text]  
• Ligon, L. A., Shelly, S. S., Tokito, M., Holzbaur, E. L.F. (2003). The Microtubule Plus-End Proteins EB1 and Dynactin Have Differential Effects on Microtubule Polymerization. Mol. Biol. Cell 14: 1405-1417 [Abstract] [Full Text]  
• Komarova, Y. A., Akhmanova, A. S., Kojima, S.-i., Galjart, N., Borisy, G. G. (2002). Cytoplasmic linker proteins promote microtubule rescue in vivo. JCB 159: 589-599 [Abstract] [Full Text]  
• Kline-Smith, S. L., Walczak, C. E. (2002). The Microtubule-destabilizing Kinesin XKCM1 Regulates Microtubule Dynamic Instability in Cells. Mol. Biol. Cell 13: 2718-2731 [Abstract] [Full Text]  
• Yvon, A.-M. C., Walker, J. W., Danowski, B., Fagerstrom, C., Khodjakov, A., Wadsworth, P. (2002). Centrosome Reorientation in Wound-Edge Cells Is Cell Type Specific. Mol. Biol. Cell 13: 1871-1880 [Abstract] [Full Text]  
• Komarova, Y. A., Vorobjev, I. A., Borisy, G. G. (2002). Life cycle of MTs: persistent growth in the cell interior, asymmetric transition frequencies and effects of the cell boundary. J. Cell Sci. 115: 3527-3539 [Abstract] [Full Text]  
• Rusan, N. M., Fagerstrom, C. J., Yvon, A.-M. C., Wadsworth, P. (2001). Cell Cycle-Dependent Changes in Microtubule Dynamics in Living Cells Expressing Green Fluorescent Protein-{alpha} Tubulin. Mol. Biol. Cell 12: 971-980 [Abstract] [Full Text]  
• Steinberg, G, Wedlich-Soldner, R, Brill, M, Schulz, I (2001). Microtubules in the fungal pathogen Ustilago maydis are highly dynamic and determine cell polarity. J. Cell Sci. 114: 609-622 [Abstract]  
• Mimori-Kiyosue, Y., Shiina, N., Tsukita, S. (2000). Adenomatous Polyposis Coli (APC) Protein Moves along Microtubules and Concentrates at Their Growing Ends in Epithelial Cells. JCB 148: 505-518 [Abstract] [Full Text]  
• Infante, A., Stein, M., Zhai, Y, Borisy, G., Gundersen, G. (2000). Detyrosinated (Glu) microtubules are stabilized by an ATP-sensitive plus-end cap. J. Cell Sci. 113: 3907-3919 [Abstract]  
• Matsumura, N., Yamazaki, T., Ihara, Y. (1999). Stable Expression in Chinese Hamster Ovary Cells of Mutated Tau Genes Causing Frontotemporal Dementia and Parkinsonism Linked to Chromosome 17 (FTDP-17). Am. J. Pathol. 154: 1649-1656 [Abstract] [Full Text]  
• Yvon, A.-M. C., Wadsworth, P., Jordan, M. A. (1999). Taxol Suppresses Dynamics of Individual Microtubules in Living Human Tumor Cells. Mol. Biol. Cell 10: 947-959 [Abstract] [Full Text]  
• Vorobjev, I., Rodionov, V., Maly, I., Borisy, G. (1999). Contribution of plus and minus end pathways to microtubule turnover. J. Cell Sci. 112: 2277-2289 [Abstract]  
• Waterman-Storer, C. M. (1998). Microtubules and Microscopes: How the Development of Light Microscopic Imaging Technologies Has Contributed to Discoveries about Microtubule Dynamics in Living Cells. Mol. Biol. Cell 9: 3263-3271 [Full Text]  
• Saoudi, Y., Fotedar, R., Abrieu, A., Doree, M., Wehland, J., Margolis, R. L., Job, D. (1998). Stepwise Reconstitution of Interphase Microtubule Dynamics in Permeabilized Cells and Comparison to Dynamic Mechanisms in Intact Cells. JCB 142: 1519-1532 [Abstract] [Full Text]  
• Liao, H., Li, Y., Brautigan, D. L., Gundersen, G. G. (1998). Protein Phosphatase 1 Is Targeted to Microtubules by the Microtubule-associated Protein Tau. J. Biol. Chem. 273: 21901-21908 [Abstract] [Full Text]  
• Panda, D., DeLuca, K., Williams, D., Jordan, M. A., Wilson, L. (1998). Antiproliferative mechanism of action of cryptophycin-52: Kinetic stabilization of microtubule dynamics by high-affinity binding to microtubule ends. Proc. Natl. Acad. Sci. USA 95: 9313-9318 [Abstract] [Full Text]  
• Cook, T. A., Nagasaki, T., Gundersen, G. G. (1998). Rho Guanosine Triphosphatase Mediates the Selective Stabilization of Microtubules Induced by Lysophosphatidic Acid. JCB 141: 175-185 [Abstract] [Full Text]  
• Gavet, O, Ozon, S, Manceau, V, Lawler, S, Curmi, P, Sobel, A (1998). The stathmin phosphoprotein family: intracellular localization and effects on the microtubule network. J. Cell Sci. 111: 3333-3346 [Abstract]  
• Charrasse, S, Schroeder, M, Gauthier-Rouviere, C, Ango, F, Cassimeris, L, Gard, D., Larroque, C (1998). The TOGp protein is a new human microtubule-associated protein homologous to the Xenopus XMAP215. J. Cell Sci. 111: 1371-1383 [Abstract]  
• Waterman-Storer, C. M., Salmon, E.D. (1997). Actomyosin-based Retrograde Flow of Microtubules in the Lamella of Migrating Epithelial Cells Influences Microtubule Dynamic Instability and Turnover and Is Associated with Microtubule Breakage and Treadmilling. JCB 139: 417-434 [Abstract] [Full Text]  
• Nabi, I. R., Guay, G., Simard, D. (1997). AMF-R Tubules Concentrate in a Pericentriolar Microtubule Domain After MSV Transformation of Epithelial MDCK Cells. J. Histochem. Cytochem. 45: 1351-1364 [Abstract] [Full Text]  
• Panda, D., Miller, H. P., Islam, K., Wilson, L. (1997). Stabilization of microtubule dynamics by estramustine by binding to a novel site in tubulin: A possible mechanistic basis for its antitumor action. Proc. Natl. Acad. Sci. USA 94: 10560-10564 [Abstract] [Full Text]  
• Keating, T. J., Peloquin, J. G., Rodionov, V. I., Momcilovic, D., Borisy, G. G. (1997). Microtubule release from the centrosome. Proc. Natl. Acad. Sci. USA 94: 5078-5083 [Abstract] [Full Text]  
• Panda, D., Singh, J. P., Wilson, L. (1997). Suppression of Microtubule Dynamics by LY290181. A POTENTIAL MECHANISM FOR ITS ANTIPROLIFERATIVE ACTION. J. Biol. Chem. 272: 7681-7687 [Abstract] [Full Text]  
• Vorobjev, I., Svitkina, T., Borisy, G. (1997). Cytoplasmic assembly of microtubules in cultured cells. J. Cell Sci. 110: 2635-2645 [Abstract]  
• Yvon, A., Wadsworth, P (1997). Non-centrosomal microtubule formation and measurement of minus end microtubule dynamics in A498 cells. J. Cell Sci. 110: 2391-2401 [Abstract]  
• Laurent, M, Fleury, A (1996). Hysteretic behavior and differential apparent stability properties of microtubule species emerge from the regulation of post-translational modifications of microtubules. J. Cell Sci. 109: 419-428 [Abstract]  
• Dhamodharan, R, Wadsworth, P (1995). Modulation of microtubule dynamic instability in vivo by brain microtubule associated proteins. J. Cell Sci. 108: 1679-1689 [Abstract]  
• Robinson, J., Vandre, D. (1995). Stimulus-dependent alterations in macrophage microtubules: increased tubulin polymerization and detyrosination. J. Cell Sci. 108: 645-655 [Abstract]  
• Bosc, C., Frank, R., Denarier, E., Ronjat, M., Schweitzer, A., Wehland, J., Job, D. (2001). Identification of Novel Bifunctional Calmodulin-binding and Microtubule-stabilizing Motifs in STOP Proteins. J. Biol. Chem. 276: 30904-30913 [Abstract] [Full Text]  
• Eichenmuller, B., Everley, P., Palange, J., Lepley, D., Suprenant, K. A. (2002). The Human EMAP-like Protein-70 (ELP70) Is a Microtubule Destabilizer That Localizes to the Mitotic Apparatus. J. Biol. Chem. 277: 1301-1309 [Abstract] [Full Text]  
• Krylyshkina, O., Kaverina, I., Kranewitter, W., Steffen, W., Alonso, M. C., Cross, R. A., Small, J. V. (2002). Modulation of substrate adhesion dynamics via microtubule targeting requires kinesin-1. JCB 156: 349-360 [Abstract] [Full Text]  

Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents