Microtubule treadmilling in vivo

Science

Volumn 275, Issue 5297, 1997, Pages 215-220

  Rodionov, Vladimir I ^a   Borisy, Gary G ^a  

^a UNIVERSITY OF WISCONSIN MADISON   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

TUBULIN;

ANIMAL CELL; ARTICLE; CENTROSOME; CYTOPLASM; DEPOLYMERIZATION; FISH; FLUORESCENCE; MELANOPHORE; MICROTUBULE ASSEMBLY; NONHUMAN; PRIORITY JOURNAL;

ANIMALS; CELL MEMBRANE; CELLS, CULTURED; CENTROSOME; CYTOPLASM; FISHES; KINETICS; MELANOPHORES; MICROTUBULES; MOVEMENT; PIGMENTS, BIOLOGICAL; POLYMERS;

ANIMALIA; MELANOPHORA;

EID: 0031013546     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.275.5297.215     Document Type: Article

References (30)

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15. V. I. Rodionov and G. G. Borisy, in preparation.
16. Melanophores of black tetra (Gymnocorymbus ternetzi) were cultured on carbon-coated glass cover slips as described previously [F. K. Gyoeva, E. V. Leonova, V. I. Rodionov, V. I. Gelfand, J. Cell Sci. 88, 649 (1987)] and injected with Cy3-labeled porcine brain tubulin prepared as described previously for X-rhodamine-labeled tubulin [ P. J. Sammak and G. G. Borisy, Nature 332, 724 (1988)].
17. Melanophores of black tetra (Gymnocorymbus ternetzi) were cultured on carbon-coated glass cover slips as described previously [F. K. Gyoeva, E. V. Leonova, V. I. Rodionov, V. I. Gelfand, J. Cell Sci. 88, 649 (1987)] and injected with Cy3-labeled porcine brain tubulin prepared as described previously for X-rhodamine-labeled tubulin [ P. J. Sammak and G. G. Borisy, Nature 332, 724 (1988)].
18. -5 M adrenalin.
19. Cover slips with attached fragments were mounted into sealed Rose chambers filled with culture medium containing 20 mM lactic acid and 2% Oxyrase to remove oxygen. MT images were captured with a charge-coupled device (CCD) camera at 3-s intervals. Acquisition of live fluorescence images of MTs was described in detail previously [V. I. Rodionov, S.-S. Lim, V. I. Gelfand, G. G. Borisy, J. Cell Biol. 126, 1455 (1994)]. Distances were measured with the use of Image-1 (Universal Imaging) software, and data were analyzed in Sigmaplot (Jandel Scientific, San Rafael, CA).
20. - for individual MTs (n = 6) showed less variation than the ratio for averages of populations of MT ends.
21. bec = 0.79. Treadmilling MTs would appear if τ < 5 min. Assuming that residence times are distributed normally, a 1 % incidence of treadmilling is consistent with a standard deviation in τ of 16 min [(43 min - 5 min)/2.3]. An average residence time of 43 ± 16 min predicts that ∼10% of the time an MT would be growing; ∼10% of the time it would be shortening; ∼80% of the time it would appear static, with one end at the pigment aggregate and the other end at the surface; and ∼1% of the time it would appear to be free and treadmilling, as observed.
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29. Video sequences of Figs. 1 and 3, as well as other sequences showing MT motion in melanophore fragments, can be seen at http://borisy.bocklabs. wisc.edu/.
30. We thank J. Peloquin for preparation of labeled tubulin. Supported by NIH grant GM25062 to G.G.B. and NSF grant MCB-9418474 to V.I.R.