Abstract
For the direct investigation of intranuclear dynamics in living cells, extremely deformed nuclei of basipetally centrifuged protonemal cells of the fernAdiantum capillus-veneris were manipulated by the laser trap and the laser scalpel. Whereas the nucleolus was tightly fixed at the central position inside the non-centrifuged nucleus and proved to be immovable by the optical trap, it could easily be trapped and moved towards three directions inside the bubble-like terminal widening of the basal thread-like extension of centrifuged nuclei. Due to the connection of the nucleolus to the chromatin inside the nuclear thread (NT), moving was not possible against the direction of the nuclear apical main body. Nucleoli in recovered nuclei were again immovable, thus indicating the presence of a dynamic nucleolar anchoring system inside the nucleus. When the nucleolus in the bubble was arrested during the thread shortening process by the optical trap, the acropetal movement of the bubble continued. Probably due to dragging forces, some nucleoli became stretched, and a thick strand of a still unknown composition stretched between the nucleolus and the insertion site of the shortening NT. To assess whether the shrinking of the nuclear envelope (NE) and the shortening of the chromatin inside the NT were independent processes, the chromatin above the bubble was cut inside the NT by the laser scalpel. After severance, a gap between the nucleolus and the end of the chromatin strand in the NT indicated the shortening of the chromatin inside the NT. From these findings it was concluded that a shortening force was existing in the chromatin of the NT and that probably no physical link existed between the chromatin and the NE.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- CW:
-
continuous wave
- DAPI:
-
4′, 6′-diamidino-2-phenylindole
- DIC:
-
differential interference contrast
- NE:
-
nuclear envelope
- NT:
-
nuclear thread
References
Adam, S.A., Sterne Marr, R. andGerace, L. 1990. Nuclear protein import in permeabilized mammalian cells requires soluble cytoplasmic factors. J. Cell Biol.111: 807–816.
Ashkin, A. andDziedzic, J.M. 1989. Internal cell manipulation using infrared laser traps. Proc. Natl. Acad. Sci. USA86: 7914–7918.
Ashkin, A., Dziedzic, J.M. andYamane, T. 1987. Optical trapping and manipulation of single cells using infrared laser beams. Nature330: 769–771.
Beams, H.W. andKessel, R.G. 1987. Development of centrifuges and their use in the study of living cells. Int. Rev. Cytol.100: 15–48.
Bourgeois, C.A. andHubert, J. 1988. Spatial relationship between the nucleolus and the nuclear envelope: structural aspects and functional significance. Int. Rev. Cytol.111: 1–52.
Bremer, J.W., Busch, H. andYeoman L.C. 1981. Evidence for a species of nuclear actin distinct from cytoplasmic and muscle actins. Biochemistry20: 2013–2017.
De Beus, E., Brockenbrough, J.S., Hong, B. andAris, J.P. 1994. Yeast NOP2 encodes an essential nucleolar protein with homology to a human proliferation marker. J. Cell Biol.127: 1799–1813.
De Boni, U. 1994. The interphase nucleus as a dynamic structure. Int. Rev. Cytol.150: 149–171.
De Boni, U. andMintz, A.H. 1986. Curvilinear, three-dimensional motion of chromatin domains and nucleoli in neuronal interphase nuclei. Science234: 863–866.
Foisner, R. andGerace, L. 1993. Integral membrane proteins of the nuclear envelope interact with lamins and chromosomes, and binding is modulated by mitotic phosphorylation. Cell73: 1267–1279.
Franke, W.W., Kleinschmidt, J.A., Spring, H., Krohne, G., Grund, C., Trendelenburg, M.F., Stoehr, M. andScheer, U. 1981. A nucleolar skeleton of protein filaments demonstrated in amplified nucleoli ofXenopus laevis. J. Cell Biol.90: 289–299.
Hahne, G. andHoffman, F. 1984. The effect of laser microsurgery on cytoplasmic strands and cytoplasmic streaming in isolated plant protoplasts. Eur. J. Cell Biol.33: 175–179.
Itoh, M. 1969. Intracellular position of nuclear division in protonema ofPteris vittata. Embryologia10: 273–283.
Kumar, A., Razluddin, Finlay, T.H., Thomas, J.O. andSzer, W. 1984. Isolation of a minor species of actin from the nuclei ofAcanthamoeba castellanii. Biochemistry23: 6753–6757.
Kurachi, M., Hoshi, M. andTashiro, H. 1995. Buckling of a single microtubule by optical trapping forces: direct measurement of microtubule rigidity. Cell Motil. Cytoskel.30: 221–228.
Ludérus, M.E.E., de Graf, A., Mattia, E., den Blaauwen, J.L., Grande, M.A., de Jong, L. andvan Driel, R. 1992. Binding of matrix attachment regions to lamin B1. Cell70: 949–959.
Mélère, T. andXue, Z. 1995. The nucleolus: an organelle formed by the act of building a ribosome. Curr. Opin. Cell Biol.7: 319–324.
Milankov, K. andDe Boni, U. 1993. Cytochemical localization of actin and myosin aggregates in interphase nucleiin situ. Exp. Cell Res.209: 189–199.
Monajembashi, S., Cremer, C., Cremer, T., Wolfrum, J. andGreulich, K.O. 1986. Microdissection of human chromosomes by a laser microbeam. Exp. Cell Res.167: 262–265
Murata, T. andWada, M. 1993. Cell division in caffeineinduced binucleate protonemal cells ofAdiantum. I. Asynchronous mitosis of two nuclei in a single cell. J. Plant Res.106: 305–311.
Ohnuki, Y., Rounds, D.E., Olson, R.S. andBerns, M.W. 1972. Laser microbeam irradiation of the juxtanucleolar regions of prophase nucleolar chromosomes. Exp. Cell Res.71: 132–144.
Rimm, D.L. andPollard, T.D. 1989. Purification and characterization of anAcanthamoeba nuclear actin-binding protein. J. Cell Biol.109: 585–591.
Sims, J.R., Karp, S. andIngber, D.E. 1992. Altering the cellular mechanical force balance results in integrated changes in cell, cytoskeletal and nuclear shape. J. Cell Sci.103: 1215–1222.
Sukegawa, J. andBlobel, G. 1993. A nuclear pore complex protein that contains zinc finger motifs, binds DNA and faces the nucleoplasm. Cell72: 29–38.
Wada, M. andFuruya, M. 1970. Photocontrol of the orientation of cell division inAdiantum. I. Effects of the dark and red periods in the apical cell of gametophytes. Dev. Growth Differ.12: 109–118.
Wada, M., Kadota, A. andFuruya, M. 1983. Intracellular localization and dichroic orientation of phytochrome in plasma membrane and/or ectoplasm of a centrifuged protonema of fernAdiantum capillus-veneris L. Plant Cell Physiol.24: 1441–1447.
Wada, M. andO'Brien, T.P. 1975. Observations on the structure of the protonema ofAdiantum capillus-veneris L. undergoing cell division following white-light irradiation. Planta126: 213–227.
Weber, G., Monajembashi, S., Geulich, K.O. andWolfrum, J. 1988. Injection of DNA into plant cells with a UV laser microbeam. Naturwissenschaften75: 35–36.
Wunsch, C. andWada, M. 1996. Recovery of interphase nuclei from extreme structural alterations in centrifuged fern protonemal cells. Eur. J. Cell Biol.71: 414–422.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Wunsch, C., Kurachi, M., Kikumoto, M. et al. Detection of intranuclear forces by the use of laser optics during the recovery process of elongated interphase nuclei in centrifuged protenemal cells ofAdiantum capillus-veneris . J. Plant Res. 111, 399–405 (1998). https://doi.org/10.1007/BF02507804
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02507804