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2 , , , ( 2+ ). 5 1 . (Isant, 1983), PtK , 2+ . 2+ 10--20 1 . . , 4--9 , . , 2+ . . 2+ -- 2 -- (0.2 ). , , . , , 2+ 2+ . , 2+ , 23187, : 30 (, , 1995). , 2+ , (Hoffstein, Weissman, 1978). , 2+ , (Marcum et al., 1978), (Welsh et al., 1979; Means, Dedman, 1980). , 2+ , . , 2+ 2 , . 2+ 10--20 , (Isant, 1983). , 2+ , (Jordan et al., 1993) (Wendell et al., 1993) , (Klienfield, Sisken, 1984; Alieva, Vorobjev, 1991). (Gunderson, Bulinski, 1988; Wadsworth, Salmon, 1988; Rieder, 1990). (De Bradander et al., 1980; Rieder, 1990). , , , . , , , (Hyman, Mitchison, 1991). , (Witt et al., 1981) (Rieder, 1981). , 1265


(, ), , . , , ( 23187 ). 2 . , . , , . , , (De Brabander et al., 1980; Rieder, 1990). , 23187 2 , . - (McDonald et al., 1992) , , . , . , . . . , . , , , . - . (. Cahalan, , , ) - . ( , , ) 2+. ( 93-04-6523 95-04-12703) ( MRJOOO).

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Hyman A. A., Mitchison . J. Two different microtubule based motor activities with opposite polarities in kinetochores // Nature. /997. Vol. 351. P. 206--211. IsantJ. G. The role of calcium ions during mitosis //Chromosoma. 1983. Vol. 88. P. 1--10. Jenses P., Rasmussen H. The effect of A23187 upon calcium metabolism in the human lymphocyte // Biochim. biophys. acta. 1977. Vol. 468. P. 146--156. Jordan M. A., Toso R. /., Thrower D., Wilson L. Mechanism of mitotic block and inhibition of cell proliferation by taxol at low concentrations // Proc. Nat. Acad. Sci. USA. /993. Vol. 90. P. 9552--9556. Kao J. P. Y., Alderton J. M., Tsien R. Y., Steinhardt R. A. Active involvement of Ca2+ in mitotic progression of Swiss 3T3 fibroblasts // J. Cell Biol. 7990. Vol. 111. P. 183--196. Keith C. H., Maxfield F. R., Shelanski M. J. Intracellular free calcium levels are reduced in mitotic PtK epithelial cells // Proc. Nat. Acad. Sci. USA. /955. Vol. 82. P. 802--804. Kiehart D. P. Studies on the in vivo sensitivity of spindle microtubules to calcium ions and evidence for a vesicular calcium-sequestering system // J. Cell Biol. 795/. Vol. 88. P. 604. Klieinfield R. G., SiskenJ. E. Morphological and kinetic aspects of mitotic arrest by and recovery from colce-mid // J. Cell Biol. 1984. Vol. 91. P. 369--379. Lewis R. S., Cahalan M. D. Mitogen-induced oscillations of cytosolic Ca and transmembrane Ca current in human leukemic T-cells //Cell Regulation. 7959. Vol. 1. P. 99--112. Marcum J. M., Dedman J. R., Brinkley B. R., Means A. R. Control of microtubule assembly-dissembly by calcium dependent regulator protein //Proc. Nat. Acad. Sci. USA. 7975. Vol. 75. P. 3771--3775. McDonaldK., O'Toole E. ., Mastronarde D. N., Mclntosh J. R. Kinetochore microtubules in PtK cells // J. Cell Biol. 7992. Vol. 118. P. 369--383. Means A. R., Dedman J. R. Calmodulin -- an intracellular calcium receptor // Nature. 7950. Vol. 285. P. 73--77. Poenie M. J., Alderton J. M., Steinhardt R. A., Tsien R. Y. Calcium rises abruptly and briefly throughout the cell at the onset of anaphase // Science. 7956". Vol. 233. P. 886--889. Poenie M. J., Alderton J. M., Tsien R. Y., Steinhardt R. A. Changes of free calcium levels with stages of the cell division cycle//Nature. 7955. Vol. 315. P. 147--149. Ratan R. R., Maxfield F. R., Shelanski M. J. Long lasting and rapid calcium changes during mitoses // J. Cell Biol. 7955. Vol. 107. P. 993--999. Rieder C. L. The structure of the cold-stable kinetochore fiber in metaphase PtK cells // Cromosoma. 7957. Vol. 84. P. 145--148. Rieder C. L. Formation of the astral mitotic spindle: ultrastructural basis for the centrosome-kinetochore interaction. //Electron. Microsc. Rev. 7990. Vol. 3. P. 269--300. Salmon E. D., Segall R. R. Calcium labile mitotic spindles isolated from sea urchin eggs (Lytechinus variega-tus) // J. Cell Biol. 7950. Vol. 86. P. 355--360. Schliwa M., Euteneuer U., Bulinski J. C., Isant J. G. Calcium-lability of cytoplasmic microtubules and its modulation by microtubule-associated proteins // Proc. Nat. Acad. Sci. USA. 7957. Vol. 78. P. 1037--1041. Silver R. ., Cole R. D., Cande W. Z. Isolation of mitotic apparatus containing vesicles with calcium sequestration activity // Cell. 7950. Vol. 19. P. 505--516. Sisken J. E., Silver R. ., Barrows G. H., Grasch S. D. Studies on the role of Ca in cell division with the use of fluorescent probes and quantitative video intensification microscopy // Advances in Microscopy. 7955. P. 73--87. Tombes R. M., Borisy G. G. Intracellular free calcium and mitosis in mammalian cells: anaphase onset is calcium modulated, but is not triggered by a brief transient // J. Cell Biol. 7959. Vol. 109. P. 627--636. Wadsworth P., Salmon E. D. Spindle microtubule dynamics: modulation by metabolic inhibitors // Cell Motil. Cytoskeleton. 7P55. Vol. 11. P. 97--105. Welsh M. J., Dedman J. R., Brinkley B. R., Means A. R. II J. Cell Biol. 7979. Vol. 81. P. 624--634. Wendell K. L., Wilson L., Jordan M. A. Mitotic block in Hela cells by vinblastine: ultrastructural changes in kinetochore-microtubule attachment and in centrosome // J. Cell Sci. 7993. Vol. 104. P. 261--274. Win P. L., Ris H., Borisy G. G. Structure of kinetochore fibers: microtubule continuity and inter-microtubule bridges // Chromosoma. 7957. Vol. 83. P. 523--540. Wolniak S. M., Bart K. M. The buffering of calcium with quin2 reversibly forestalls anaphase onset in stamen hair cells of Tradescantia // Eur. J. Cell Biol. 7955. Vol. 39. P. 33--40. Ziegler M. L., SiskenJ. E., Vedbrat S. The alteration of mitotic events by ionophore A23187 and carbonnyl cyanide n-chlorophenylhydrazone // J. Cell Sci. 7955. Vol. 75. P. 347--355. 15 X 1995

THE EFFECT OF [Ca++] RISE ON MITOTIC PK CELLS TREATED WITH CALCIUM IONOPHORE A23187 7. B. Alieva, I. A. Vorobjev
Institute of Physical and Chemical Biology, Moscow University

After addition of 20 mM calcium ionophore A23187 to cultured PK (pig kidney embryo) cells, [Ca++] in cytosol increased by more than 10 times. The maximum [Ca++] concentration was observed 1--2 min after drug introduction. Later on [Ca++] gradually decreased, and after 30 min of incubation with A23187 [Ca++] its concentration was 1267


3--5 times higher than in the norm. 1 min after introduction of the calcium ionophore, mitotic spindles shortened for 1/3 and the angle of divergence of spindle microtubules from the centrosome extended. These changes remained for 5 min of treatment. After nocodazole treatment the length of the mitotic spindle reduced (2 min), then mitotic spindle and the metaphase plate were disrupted. The rate of mitotic spindle shortening after addition of the ionophore is about the same as after addition of nocodazole, but after ionophore treatment the metaphase plate remained for more than 5 min. Based on the results obtained we suggest that the maximum distance between spindle poles at metaphase is in the intact cell, and after any perturbation of normal microtubule dynamics the spindle may rapidly collapse. The collapsed mitotic spindle becomes more stable and its size is determined predominantly by kinetochore fibers. The metaphase spindle is completely and rapidly destroyed when the microtubule growth is prohibited, but it is preserved when this growth is restricted.


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