Abstract
Cells were prepared for indirect immunofluorescence microscopy after paraformaldehyde fixation of multicellular root apices and brief incubation in cell wall-digesting enzymes. This allowed subsequent separation of the tissue into individual cells or short files of cells which were put onto coverslips coated with polylysine. Unlike spherical protoplasts made from living tissues, these preparations retain the same polyhedral shape as the cells from which they are derived. Cellular contents, including organized arrays of microtubules, are likewise structurally stabilized. Antibodies to porcine brain tubulin react with all types of microtubule array known to occur in plant meristematic cells, namely, interphase cortical microtubules, pre- prophase bands, the mitotic spindle, and phragmoplast microtubules. The retention of antigenicity in permeabilized, isolated, stabilized cells from typical, wall-enclosed plant cells has much potential for plant immunocytochemistry, and in particular should facilitate work on the role of microtubules in the morphogenesis of organized plant tissues.
Full Text
The Full Text of this article is available as a PDF (801.9 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Brinkley B. R., Fistel S. H., Marcum J. M., Pardue R. L. Microtubules in cultured cells; indirect immunofluorescent staining with tubulin antibody. Int Rev Cytol. 1980;63:59–95. doi: 10.1016/s0074-7696(08)61757-x. [DOI] [PubMed] [Google Scholar]
- Carpita N., Sabularse D., Montezinos D., Delmer D. P. Determination of the pore size of cell walls of living plant cells. Science. 1979 Sep 14;205(4411):1144–1147. doi: 10.1126/science.205.4411.1144. [DOI] [PubMed] [Google Scholar]
- Franke W. W., Seib E., Herth W., Osborn M., Weber K. Reaction of the anastral mitotic apparatus of endosperm cells of the plant Leucojum aestivum with antibodies to tubulin from porcine brain as revealed by immunofluorescence microscopy. Cell Biol Int Rep. 1977 Jan;1(1):75–83. doi: 10.1016/0309-1651(77)90013-3. [DOI] [PubMed] [Google Scholar]
- Gunning B. S. Spatial and temporal regulation of nucleating sites for arrays of cortical microtubules in root tip cells of the water fern Azolla pinnata. Eur J Cell Biol. 1980 Dec;23(1):53–65. [PubMed] [Google Scholar]
- Hardham A. R., Gunning B. E. Structure of cortical microtubule arrays in plant cells. J Cell Biol. 1978 Apr;77(1):14–34. doi: 10.1083/jcb.77.1.14. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hepler P. K., Jackson W. T. Microtubules and early stages of cell-plate formation in the endosperm of Haemanthus katherinae Baker. J Cell Biol. 1968 Aug;38(2):437–446. doi: 10.1083/jcb.38.2.437. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hepler P. K., Newcomb E. H. Fine structure of cell plate formation in the apical meristem of Phaseolus roots. J Ultrastruct Res. 1967 Aug 30;19(5):498–513. doi: 10.1016/s0022-5320(67)80076-5. [DOI] [PubMed] [Google Scholar]
- KLEIN S., GINZBURG B. An electron microscopic investigation into the effect of EDTA on plant cell wall. J Biophys Biochem Cytol. 1960 Apr;7:335–338. doi: 10.1083/jcb.7.2.335. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Knox R. B., Vithanage H. I., Howlett B. J. Botanical immunocytochemistry: a review with special reference to pollen antigens and allergens. Histochem J. 1980 May;12(3):247–272. doi: 10.1007/BF01006951. [DOI] [PubMed] [Google Scholar]
- LETHAM D. S. The separation of plant cells with ethylenediaminetetraacetic acid. Exp Cell Res. 1960 Nov;21:353–360. doi: 10.1016/0014-4827(60)90267-6. [DOI] [PubMed] [Google Scholar]
- Marchant H. J. Microtubules associated with the plasma membrane isolated from protoplasts of the green alga Mougeotia. Exp Cell Res. 1978 Aug;115(1):25–30. doi: 10.1016/0014-4827(78)90397-x. [DOI] [PubMed] [Google Scholar]
- Osborn M., Weber K. The display of microtubules in transformed cells. Cell. 1977 Nov;12(3):561–571. doi: 10.1016/0092-8674(77)90257-4. [DOI] [PubMed] [Google Scholar]
- Pickett-Heaps J. D., Northcote D. H. Cell division in the formation of the stomatal complex of the young leaves of wheat. J Cell Sci. 1966 Mar;1(1):121–128. doi: 10.1242/jcs.1.1.121. [DOI] [PubMed] [Google Scholar]
- Pickett-Heaps J. D., Northcote D. H. Organization of microtubules and endoplasmic reticulum during mitosis and cytokinesis in wheat meristems. J Cell Sci. 1966 Mar;1(1):109–120. doi: 10.1242/jcs.1.1.109. [DOI] [PubMed] [Google Scholar]
- Weber K., Wehland J., Herzog W. Griseofulvin interacts with microtubules both in vivo and in vitro. J Mol Biol. 1976 Apr 25;102(4):817–829. doi: 10.1016/0022-2836(76)90293-x. [DOI] [PubMed] [Google Scholar]