Summary
Myosin-related proteins have been localized immunocytochemically in gravity-sensing rhizoids of the green algaChara globularis using a monoclonal antibody against the heavy chain of myosin from mouse 3T3 cells and a polyclonal antibody to bovine skeletal and smooth muscle myosin. In the basal zone of the rhizoids which contain a large vacuole, streaming endoplasm and stationary cortical cytoplasm, the monoclonal antibody stained myosin-related proteins as diffusely fluorescing endoplasmic strands. This pattern is similar to the arrangement of subcortical actin filament bundles. In the apical zone which contains an aggregation of ER membranes and secretory vesicles for tip growth, diffuse immunofluorescence was detected; the intensity of the signal increasing towards the apical cell wall. The most prominent myosin-staining was associated with the surface of statoliths in the apical zone. The polyclonal antibody produced a punctate staining pattern in the basal zone, caused by myosin-related proteins associated with the surface of drganelles in the streaming endoplasm and the periphery of the nucleus. In the apical zone, this antibody revealed myosin-immunofluorescence on the surface of statoliths in methacrylate-embedded rhizoids. Neither antibody revealed myosin-immunofluorescence on the surface of organelles and vesicles in the relatively stationary cytoplasm of the subapical zone. These results indicate (i) that different classes of myosin are involved in the various transport processes inChara rhizoids; (ii) that cytoplasmic streaming in rhizoids is driven by actomyosin, corresponding to the findings onChara internodal cells; (iii) that actindependent control of statolith position and active movement is mediated by myosin-related proteins associated with the statolith surfaces; and (iv) that myosin-related proteins are involved in the process of tip growth.
Similar content being viewed by others
References
Baines IC, Brzeska H, Korn ED (1992) Differential localization ofAcanthamoeba myosin I isoforms. J Cell Biol 119: 1193–1203
Bartnik E, Sievers A (1988) In-vivo observation of a spherical aggregate of endoplasmic reticulum and of Golgi vesicles in the tip of fast-growingChara rhizoids. Planta 176: 1–9
—, Hejnowicz Z, Sievers A (1990) Shuttle-like movements of Golgi vesicles in the tip of growingChara rhizoids. Protoplasma 159: 1–8
Baskin TL, Busby CH, Fowke LC, Sammut M, Gubler F (1992) Improvement in immunostaining samples embedded in methacrylate: localization of microtubules and other antigens throughout developing organs in plants of diverse taxa. Planta 187: 405–413
Braun M, Sievers A (1993) Centrifugation causes adaptation of microfilaments. Studies on the transport of statoliths in gravity sensingChara rhizoids. Protoplasma 174: 50–61
— — (1994) Role of the microtubule cytoskeleton in gravisensingChara rhizoids. Eur J Cell Biol 63: 289–298
Buchen B, Braun M, Hejnowicz Z, Sievers A (1993) Statoliths pull on microfilaments. Experiments under microgravity. Protoplasma 172: 38–42
Conrad PA, Giuliano KA, Fisher G, Collins K, Matsudaira PT, Taylor DL (1993) Relative distribution of actin, myosin I, and myosin II during the wound healing response of fibroblasts. J Cell Biol 120: 1381–1391
Cresti M, Ciampolini F, Mulcahy DL, Mulcahy G (1985) Ultrastructure ofNicotiana alata pollen, its germination and early tube formation. Am J Bot 72: 719–727
Forsberg C (1965) Nutritional studies ofChara in axenic cultures. Physiol Plant 18: 275–290
Fukui Y, Lynch TJ, Brzeska H, Korn ED (1989) Myosin I is located at the leading edge of locomotingDictyostelium amoebae. Nature 341: 328–331
Grolig F, Williamson RE, Parke J, Miller C, Anderton BH (1988) Myosin and Ca2+-sensitive streaming in the algaChara; detection of two polypeptides reacting with monoclonal anti-myosin and their localization in the streaming endoplasm. Eur J Cell Biol 47: 22–31
Hejnowicz Z, Sievers A (1981) Regulation of the position of statoliths inChara rhizoids. Protoplasma 108: 117–137
—, Buchen B, Sievers A (1985) The endogenous difference in the rates of acropetal and basipetal cytoplasmic streaming inChara rhizoids is enhanced by gravity. Protoplasma 125: 219–229
Heslop-Harrison J, Heslop-Harrison Y (1989) Myosin associated with the surface of organelles, vegetative nuclei and generative cells in angiosperm pollen grains and tubes. J Cell Sci 94: 319–325
Kachar B, Reese TS (1988) The mechanism of cytoplasmic streaming in characean algal cells: sliding of endoplasmic reticulum along actin filaments. J Cell Biol 106: 1545–1552
Kato T, Tonomura Y (1977) Identification of myosin inNitella flexilis. J Biochem 82: 777–782
Kohno T, Shimmen T (1988a) Accelerated sliding of pollen tube organelles along Characeae actin bundles regulated by Ca2+. J Cell Biol 91: 501–509
— — (1988b) Mechanism of Ca2+ inhibition of cytoplasmic streaming in lily pollen tubes. J Cell Sci 91: 501–509
Kuroda K (1990) Cytoplasmic streaming in plant cells. Int Rev Cytol 121: 267–307
Lancelle SA, Cresti M, Hepler PK (1987) Ultrastructure of the cytoskeleton in freeze-substituted pollen tubes ofNicotiana alata. Protoplasma 140: 141–150
Leitz G, Schnepf E, Greulich KO (1995) Micromanipulation of Statoliths in gravity-sensingChara rhizoids by optical tweezers. Planta 197: 278–288
Ma Y, Yen LF (1989) Actin and myosin in pea tendrils. Plant Physiol 89: 586–589
Miller DD, Scordilis SP, Hepler PK (1995) Identification and localization of three classes of myosin in pollen tubes ofLilium longiflorum andNicotiana alata. J Cell Sci 108: 2549–2563
Nagai R (1993) Regulation of intracellular movements in plant cells by environmental stimuli. Int Rev Cytol 145: 251–310
Ohsuka K, Inoue A (1979) Identification of myosin in a flowering plant,Egeria densa. J Biochem 85: 375–378
Parke JM, Miller JC, Anderton BH (1986) Higher plant myosin heavy-chain identified using a monoclonal antibody. Eur J Cell Biol 41: 9–13
Picton JM, Steer MW (1981) Determination of secretory vesicles production rates by dictyosomes in pollen tubes ofTradescantia using cytochalasin. J Cell Biol 49: 261–272
Qiao L, Grolig F, Jablonsky PP, Williamson RE (1989) Myosin heavy chains: detection by immunoblotting in higher plants and localization by immunofluorescence in the algaChara. Cell Biol Int Rep 13: 107–117
Shimmen T, Tazawa M (1982) Reconstitution of cytoplasmic streaming in characean algae. Protoplasma 113: 127–131
Sievers A, Kramer-Fischer M, Braun M, Buchen B (1991a) The polar organization of the growingChara rhizoid and the transport of statoliths are actin-dependent. Bot Acta 104: 103–109
—, Buchen B, Volkmann D, Hejnowicz Z (1991b) Role of the cytoskeleton in gravity perception. In: Lloyd CW (ed) The cytoskeletal basis of plant growth and form. Academic Press, London, pp 169–182
- Braun M, Hejnowicz Z (1994) Gravity and the cytoskeleton. In: Proceedings of the Fifth European Symposium on Life Sciences Research in Space, Arcachon, France, 26 Sept–1 Oct 1993, ESA SP-366, Nordwijk, pp 15–17
Tang X, Hepler PK, Scordilis SP (1989) Immunochemical and immunocytochemical identification of a myosin heavy chain polypeptide inNicotiana pollen tubes. J Cell Sci 92: 569–574
Tewinkel M, Kruse S, Quader H, Volkmann D, Sievers A (1989) Visualization of actin filament pattern in plant cells without prefixation. A comparison of differently modified phallotoxins. Protoplasma 149: 178–182
Tirlapur UK, Cai G, Faleri C, Moscatelli A, Scali M, DelCasino C, Tiezzi A, Cresti M (1995) Confocal imaging and immunogold electron microscopy of changes in distribution of myosin during pollen hydration, germination and pollen tube growth inNicotiana tabacum L. Eur J Cell Biol 67: 209–217
Vahey M, Titus M, Trautwein R, Scordilis S (1982) Tomato actin and myosin: contractile proteins from a higher plant. Cell Motil 2: 131–148
Volkmann D, Czaja AWP (1981) Reversible inhibition of secretion in root cap cells of cress after treatment with cytochalasin B. Exp Cell Res 135: 229–236
—, Buchen B, Hejnowicz Z, Tewinkel M, Sievers A (1991) Oriented movement of statoliths studied in a reduced gravitational field during parabolic flights of rockets. Planta 185: 153–161
Wasteneys GO, Williamson RE (1991) Endoplasmic microtubules and nucleus-associated actin rings inNitella internodal cells. Protoplasma 162: 86–98
Wunsch C, Volkmann D (1993) Immunocytological detection of myosin in the root tip cells ofLepidium sativum. Eur J Cell Biol 60 [Suppl 37]: 64
Yokota E, Shimmen T (1994) Isolation and characterization of plant myosin from pollen tubes of lily. Protoplasma 177: 153–162
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Braun, M. Immunolocalization of myosin in rhizoids ofChara globularis Thuill. Protoplasma 191, 1–8 (1996). https://doi.org/10.1007/BF01280820
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01280820