Abstract
Calcium-containing solutions were microinjected into dividing PtK1 cells to assess the effect of calcium ion concentration on the morphology and physiology of the mitotic spindle. Solutions containing 50 μM or more CaCl2 are immediately and irreversibly toxic to PtK1 cells. Those containing 5–10 μM CaCl2 cause reversible reduction in spindle birefringence followed by normal anaphase and cytokinesis. Microinjection of 5 μM or less CaCl2 into anaphase PtK1 cells has no detectable effect on the rate or extent of chromosome movement. Metaphase cells tend to enter anaphase 4–5 min after injection with 1–10 μM CaCl2, compared with an average of 16 min after injection with calcium-free buffer. Reducing the intracellular calcium concentration by injection of EGTA-CaCl2 buffers increases the lag between injection and anaphase to 20 min or more. Microinjection of calcium solutions does not promote precocious chromatid separation in nocodazole-arrested metaphase cells, indicating that the increase in calcium concentration does not induce centromere separation directly. An increase in the concentration of free calcium ions during metaphase appears to stimulate the onset of anaphase. Such an increase, regulated by the cell itself, may contribute to the initiation of chromosome separation in mammalian cells.
Similar content being viewed by others
References
Anderson B, Osborn M, Weber K (1978) Specific visualization of the distribution of the calcium-dependent regulatory protein of cyclic nucleotide phosphodiesterase (modulator protein) in tissue culture cells by immunofluorescence microscopy: mitosis and intercellular bridge. Cytobiologie 17:354–364
Berkowitz SA, Wolff (1981) Intrinsic calcium sensitivity of tubulin polymerization. The contributions of temperature, tubulin concentration, and associated proteins. J Biol Chem 256:11216–11223
Blum JJ, Hayes A, Jamieson GA, Vanamon TC (1980) Calmodulin confers calcium sensitivity on ciliary dynein ATPase. J Cell Biol 87:386–397
Bygrave FL (1978) Mitochondria and the control of intracellular calcium. Biol Rev 53:43–79
Cande WZ (1981) Physiology of chromosome movement in lysed cell models. In: Schweiger H.G., ed, International cell biology, 1980–81, Springer, Berlin·Heidelberg·New York, pp 382–391
Cande WZ (1982) Inhibition of spindle elongation in permeabilized mitotic cells by erythro-9-[3-(2-hydroxynonyl)]adenine. Nature 295:700–701
Cande WZ, Wolniak SM (1978) Chromosome movement in lysed mitotic cells is inhibited by vanadate. J Cell Biol 79:573–580
Carafoli E, Crompton M (1976) Calcium ions and mitochondria. Symp Soc Exp Biol 30:89–115
Fuller GM, Brinkley BR (1976) Structure and control of assembly of cytoplasmic microtubules in normal and transformed cells. J Supramol Struct 5:497–514
Goldstein DA (1979) Calculation of the concentrations of free cations and cation-like complexes in solutions containing multiple divalent cations and ligands. Biophys J 26:235–242
Goldstein LSB (1981) Kinetochore structure and its role in chromosome orientation during the first meiotic division in male D. melanogaster. Cell 25:591–602
Hallet MB, Campbell AK (1982) Measurement of changes in cytoplasmic free Ca2+ in fused cell hybrids. Nature 295:155–158
Hepler PK (1980) Membranes in the mitotic apparatus of barley cells. J Cell Biol 86:490–499
Hyams J (1982) Dynein in the spindle? Nature 295:648–649
Inoue S (1981) Video image processing greatly enhances contrast, quality and speed in polarization-based microscopy. J Cell Biol 89:346–356
Inoué S, Sato H (1967) Cell motility by labile association of molecules. The nature of mitotic spindle fibers and their role in chromosome movement. J Gen Physiol 50:259–292
Izant JG, Weatherbee JA, McIntosh JR (1983) A microtubule-associated protein antigen unique to the mitotic spindle in PtK1 cells. J Cell Biol 96:424–434
Jamieson GA, Vanamon TC, Blum JJ (1979) Presence of calmodulin in Tetrahymena. Proc Natl Acad Sci 76:6471–6475
Job D, Rauch CT, Fisher EH, Margolis RL (1981) Ca2+-calmodulin and protein kinase regulation of brain microtubule cold-stability. J Cell Biol 91:322a
Keller TCS III, Jemiolo DK, Burgess WH, Rebhun LI (1982) Strongylocentrotus purpuratus spindle tubulin II. Characteristics of its sensitivity to Ca++ and the effects of calmodulin isolated from bovine brain and S. purpuratus eggs. J Cell Biol 93:797–803
Kiehart DP (1981) Studies on the in vivo sensitivity of spindle microtubules to calcium ions and evidence for a vesicular calcium-sequestering system. J Cell Biol 88:604–617
Luby-Phelps K, Porter KR (1982) The control of Pigment migration in isolated erythrophores of Holoceutrus ascensionis (Osbeck) II. The role of calcium. Cell 292:441–450
Marcum JM, Dedman JR, Brinkley BR, Means AR (1978) Control of microtubule assembly-disassembly by calcium dependent regulator protein. Proc Natl Acad Sci 75:3771–3775
Mazia D, Petzelt C, Williams RO, Meza I (1972) A calcium activated ATPase in the mitotic apparatus of the sea urchin (isolated by a new method). Exp Cell Res 70:325–332
McIntosh JR, Landis SC (1971) The distribution of spindle micro-tubules during mitosis in cultured human cells. J Cell Biol 49:468–497
Means AR, Dedman JR (1980) Calmodulin — an intracellular calcium receptor. Nature 285:73–77
Mohri H, Mabuchi I, Ogawa K, Kuriyama R, Sakai H (1976) Evidence for participation of dynein in chromosome movement in mitosis. In: Perry SV, Margreth A, Adelstein RS, eds. Contractile systems in non-muscle cells, North-Holland, New York
Mueller C, Graessmann M, Graessmann A (1981) A microinjection technique converting living cells into test tubes. In: Schweiger HG, ed. International cell biology, 1980–81, Springer, Berlin-Heidelberg-New York, pp 119–127
Nishida E, Kumagai H (1980) Calcium sensitivity of sea urchin tubulin in in vitro assembly and the effects of calcium-dependent regulator (CDR) proteins isolated from sea urchin eggs and porcine brains. J Biochem 87:143–151
Olmsted JB, Borisy GG (1975) Ionic and nucleotide requirements for microtubule polymerization in vitro. Biochemistry 14:2996–3005
Pratt MM, Otter T, Salmon ED (1980) Dynein-like Mg+2-ATPase in mitotic spindles isolated from sea urchin embryos (Strongylocentrotus droebachiensis). J Cell Biol 86:738–745
Rasmussen H, Clayberger C, Gustin M (1979) The messenger function of calcium in cell activation. Symp Soc Exp Biol 33:161–197
Rebhun LI (1977) Cyclic nucleotides, calcium and cell division. Int Rev Cytol 49:1–54
Rebhun LI, Jemiolo D, Keller T, Burgess W, Kretsinger R (1980) Calcium, calmodulin and control of assembly of brain and spindle microtubules. In: DeBrabander, DeMey, eds. Microtubules and microtubule inhibitors, Elsevier/North-Holland
Roos UP (1973) Light and electron microscopy of rat kangaroo cells in mitosis. II. Kinetochore structure and function. Chromosoma 41:198–220
Rosenfeld AC, Zackroff RV, Weisenberg (1976) Magnesium stimulation of calcium binding to tubulin and calcium-induced depolymerization of microtubules. FEBS Lett 65:144–147
Roth LE, Daniels EW (1962) Electron microscopic studies of mitosis in amoebae. J Cell Biol 12:57–78
Salmon ED (1975) Pressure-induced depolymerization of spindle microtubules: I Changes in birefringence and spindle length. J Cell Biol 65:603–614
Salmon ED (1982) Calcium, spindle microtubule dynamics and chromosome movement. Cell Differ 11:353–355
Salmon ED, Segall RR (1980) Calcium-labile mitotic spindles isolated from sea urchin eggs (Lytechinus variegatus). J Cell Biol 86:355–365
Salmon ED, McKeel M, Hays T (1982) The rapid rate of tubulin dissociation from microtubules in the mitotic spindle in vivo. J Cell Biol 95:309a
Schliwa M (1976) The role of divalent cations in the regulation of microtubule assembly in vivo studies on microtubules of the heliozoan axopodium using the ionophore A23187. J Cell Biol 70:527–540
Schliwa M (1981) Proteins associated with cytoplasmic actin. Cell 25:587–590
Schliwa M, Euteneur U, Bulinski JC, Izant JG (1981) Calcium lability of cytoplasmic microbutules and its modulation by microtubule-associated proteins. Proc Natl Acad Sci 78:1037–1041
Sillen LG, Martell A (1964) Stability constants of metal-ion complexes, 2nd Edition, Chemical Society, London
Silver RB, Cole RD, Cande WZ (1980) Isolation of mitotic apparatus containing vesicles with calcium sequestration activity. Cell 19:505–516
Sisken JE, VedBrat SS (1977) On the effects of variations in intracellular and extracellular calcium ions on mitosis and cytokinesis of HeLa cells. J Cell Biol 75:263a
Tsien RY, Pozzau T, Rink TJ (1982) T-cell mitogens cause early changes in cytoplasmic free Ca2+ and membrane potential in lymphocytes. Nature 295:68–71
Weisenberg RC (1972) Microtubule formation in vitro in solutions containing low calcium concentrations. Science 177:1104–1105
Welsh MJ, Dedman JR, Brinkley BR, Means AR (1978) Calcium-dependent regulator protein: Localization in mitotic apparatus of eukaryotic cells. Proc Natl Acad Sci 75:1867–1871
Welsh MJ, Dedman JR, Brinkley BR, Means AR (1979) Tubulin and calmodulin. Effects of microtubule and microfilament inhibitors on localization in the mitotic apparatus. J Cell Biol 81:624–634
Wick SM, Hepler PK (1980) Localization of Ca++-containing antimonate precipitates during mitosis. J Cell Biol 86:500–513
Wolniak SM, Hepler PK, Jackson WT (1980) Detection of the membrane-calcium distribution during mitosis in Haemauthus endosperm with chlorotetracycline. J Cell Biol 87:23–32
Wolniak SM, Hepler PK, Jackson WT (1981) Ionic changes in the mitotic apparatus during the metaphase/anaphase transition. J Cell Biol 91:313a
Zirkle RE (1970) Involvement of the prometaphase kinetochore in prevention of precocious anaphase. J Cell Biol 47:235a
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Izant, J.G. The role of calcium ions during mitosis. Chromosoma 88, 1–10 (1983). https://doi.org/10.1007/BF00329497
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00329497