Summary
Detergent extracted spermatozoids of the fernPteridium aquilinum were used as mixed antigen preparations for raising monoclonal antibodies in order to obtain reagents for detecting as yet uncharacterized components of the plant cytoskeleton. Selected antibodies were studied by immunofluorescence microscopy of developing spermatids and mature spermatozoids. Some reacted directly with fixed cells, others required permeabilization treatments with cold methanol or Triton X-100. AntibodiesPas2D9 andPas6D7 bind to glycoprotein antigenic determinants that are exposed on the surface of the plasma membrane. Several antibodies interact with cytoskeletal components.Pas1D3,Pas5D8 andPas5F4 bind to the cytoskeleton of permeabilized cells including the flagella. Three react specifically with the flagellar band or associated components:Pas2G6 reacts with the whole flagellar band but shows a prominent binding to basal bodies,Pas5E2 binds exclusively to basal bodies, andPas5E7 detects mitochondria associated with the flagellar band. Cross-reactions to wheat root tip cells at different stages of the cell cycle are described inMarc andGunning (1988).
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Abbreviations
- MLS:
-
multilayered structure
- MT:
-
microtubule
- MAb:
-
monoclonal antibody
- MAP:
-
microtubule associated protein
References
Bell PR (1974 a) The origin of the multilayered structure in the spermatozoid ofPteridium aquilinum. Cytobiologie 8: 203–212
— (1974 b) Microtubules in relation to flagellogenesis inPteridium spermatozoids. J Cell Sci 15: 99–111
— (1979) Gametogenesis and fertilization in ferns. In:Dyer AF (ed) The experimental biology of ferns. Academic Press, London, pp 471–503
—,Duckett JG (1976) Gametogenesis and fertilization inPteridium. Bot J Linnean Soc 73: 47–78
Bloodgood RA, Woodward MP, Salomonsky NL (1986) Redistribution and shedding of flagellar membrane glycoproteins visualized using an anti-carbohydrate monoclonal antibody and Concanavalin A. J Cell Biol 102: 1797–1812
Clayton L, Black CM, Lloyd CW (1985) Microtubule nucleating sites in higher plant cells identified by an auto-antibody against pericentriolar material. J Cell Biol 101: 319–324
Connolly JA, Kalnins VI (1978) Visualization of centrioles and basal bodies by fluorescent staining with nonimmune rabbit sera. J Cell Biol 79: 526–532
Duckett JG (1975) Spermatogenesis in pteridophytes. In:Duckett JG, Racey PA (eds) The biology of the male gamete. Academic Press, London. (Biol J Linnean Soc Suppl 17: 97–127)
—,Carothers ZB (1982) The assembly of flagella during spermatogenesis in land plants. In:Amos WB, Duckett JG (eds) Prokaryotic and Eukaryotic flagella. Cambridge University Press, Cambridge. (Proc Exp Biol Symp 35, pp 533–561)
—,Klekowski EJ Jr, Hickok LG (1979) Ultrastructural studies of mutant spermatozoids in ferns. I. The mature nonmotile spermatozoid of mutation 230X inCeratopteris thalictroides (L) Brongn. Gamete Res 2: 317–343
Fais DA, Nadezhdina ES, Chentsov YS (1986) The centriolar rim. The structure that maintains the configuration of centrioles and basal bodies in the absence of their microtubules. Exp Cell Res 164: 27–30
Geiger B (1983) Membrane-cytoskeleton interaction. Biochim Biophys Acta 737: 305–341
Giloh H, Sedat JW (1982) Fluorescence microscopy: Reduced photobleaching of rhodamine and fluorescein protein conjugates by n-propyl gallate. Science 217: 1252–1255
Lancelle SA, Callaham DA, Hepler PK (1986) A method for rapid freezing of plant cells. Protoplasma 131: 153–165
Lin W, Fung B, Shyamala M, Kasamatsu H (1981) Identification of antigenically related polypeptides at centrioles and basal bodies. Proc Nat Acad Sci USA 78: 2372–2377
Marc J, Gunning BES (1986) Immunofluorescent localization of cytoskeletal tubulin and actin during spermatogenesis inPteridium aquilinum (L) Kuhn. Protoplasma 134: 163–177
— — (1988) Monoclonal antibodies to a fern spermatozoid detect novel components of the mitotic and cytokinetic apparatus in higher plant cells. Protoplasma 142: 15–24
Marchesi VT (1985) Stabilizing infrastructure of cell membranes. Ann Rev Cell Biol 1: 531–561
Oi VT, Herzenberg LA (1980) Immunoglobulin-producing hybridoma cell lines. In:Mishell BB, Shiigi SM (eds) Selected Methods in Cellular Immunology. Freeman, San Francisco, pp 351–372
Palevitz BA (1987) Actin in the preprophase band ofAllium cepa. J Cell Biol 104: 1515–1519
Stone IG (1961) The gametophytes of the Victorian Blechnaceae. I.Blechnum nudum (Labill) Luerss. Aust J Bot 9: 20–36
Tiwari SC, Wick SM, Williamson RE, Gunning BES (1984) Cytoskeleton and integration of cellular function in cells of higher plants. J Cell Biol 99: 63s-69s
Turksen K, Aubin JE, Kalnins VI (1982) Identification of a centriole-associated protein by antibodies present in normal rabbit sera. Nature (London) 298: 763–765
Wick SM (1985) The higher plant mitotic apparatus: redistribution of microtubules, calmodulin and microtubule initiation material during its establishment. Cytobios 43: 285–294
—,Muto S, Duniec J (1985) Double immuno-fluorescence labelling of calmodulin and tubulin in dividing plant cells. Protoplasma 126: 198–206
Wolniak SM, Cande WZ (1980) Physiological requirements for ciliary reactivation of bracken fern spermatozoids. J Cell Sci 43: 195–207
Woodward MP, Young WW Jr, Bloodgood RA (1985) Detection of monoclonal antibodies specific for carbohydrate epitopes using periodate oxidation. J Immunol Methods 78: 143–153
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Marc, J., Gunning, B.E.S., Hardham, A.R. et al. Monoclonal antibodies to surface and cytoskeletal components of the spermatozoid ofPteridium aquilinum . Protoplasma 142, 5–14 (1988). https://doi.org/10.1007/BF01273221
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DOI: https://doi.org/10.1007/BF01273221