FITC-labeled lectins and calcofluor white ST as probes for the investigation of the molecular architecture of cell surfaces. Studies on conjugatophycean species

Sengbusch, Peter von; Mix, Marianne; Wachholz, Irmhild; Manshard, Elsbeth

doi:10.1007/bf01287645

Cited by 31 publications

(20 citation statements)

References 33 publications

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“…Cellular expression of certain glycoconjugates depends on the developmental and physiological states as illustrated in algal systems (Surek & Sengbusch 1981, Sengbusch et al 1982. Seagrass walls are clearly rich in carbohydrates that provide general cell wall functions including structural integrity and, combined with the plasmalemma, regulation and exchange of solutes between the cytoplasm and external environment (Waldron et al 1989, Kuo et al 1990).…”

Section: Discussionmentioning

confidence: 99%

“…Distributions of glycoconjugates on cell surfaces have been demonstrated for species in a variety of algal divisions: the Cyanophyta, Pyrrhophyta, Raphidophyta, Euglenophyta, Chromophyta and Chloro-phyta (Surek & Sengbusch 1981, Vanni et al 1981, Sengbusch et al 1982, Sengbusch & Muller 1983, Imam et al 1984, Kaska et al 1988. In unicellular green algae, glycoconjugates are involved in cell-tocell recognition, adhesion, morphogenesis and wall assembly (Imam et al 1984, Klis et al 1985, Goodenough et al 1986, Schlipfenbacher et al 1986, Samson et al 1987.…”

Section: Introductionmentioning

confidence: 99%

“…Specific glycoconjugates on algal cell surfaces have been detected via light microscopy and the use of a fluorescent chromophore to label the lectin. Emissions from the cbromophores have been visualized with epifluorescence microscopy and have localized discrete domains of probe binding on algal surfaces (Surek & Sengbusch 1981, Vanni et al 1981, Sengbusch et al 1982, Sengbusch & Muller 1983.…”

Section: Introductionmentioning

confidence: 99%

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Lectins probe molecular films in biofouling:characterization of early films on non-living and living surfaces

Michael¹,

Smith²

1995

Mar. Ecol. Prog. Ser.

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Films that form quickly on surfaces immersed in the ocean influence the settlement of microbes and eukaryotic cells via differential distribu.tion of specific receptors that initiate attachment and metamorphosis. This study probed both inert, non-biological surfaces (glass slides) and living surfaces (leaves of seagrass Halophila hawaiiana) with fluorescently labelled lectins to detect and describe distributions of glycoconjugates deployed on surface films. Lectins from Canavalis ensiformis (Con A) and Limulus polyphemus (limulin) bound particular glycoconjugates in films that formed 1 and 3 d follo\ving immersion of glass slides in Pearl Harbor (Hawai'i, USA) waters. A complex spatial pattern in film development was observed on glass surfaces; approximately circular areas of reduced receptor densit~es were interspersed in an otherwise homogeneous matrix. The circular area surrounded centrally located aggregates of particles (including microbes) in 1 d films. The diameter of these zones increased nearly 10-fold when 1 and 3 d films \yere compared. A third lectin from Helix pomatia localized a third set of glycoconjugates in specific filmed areas and with the organic matter associated with bacteria on 3 d films. Each lectin also bound to surface glycoconjugates on newly emergent leaves of H. hawaiiana with qualitatively different results than those observed for glass. The distributions of receptors for Con A, H. pomatia and limulin lectins were less dense on seagrass surfaces than on filmed glass. Also, receptors for limulin and H. pomatia lectins were associated with the cell walls and peripheral cytoplasm of seagrass cells, not in surface films on leaves. Con A localized glycoconjugates showed only a faint signal on the cell walls of these young leaves. This technique documents previously undescribed spatial and chemical heterogeneities of early, similarly aged surfaces. This understanding allows the definition and localization of possible microscale cues for biofouling. These early steps are expected to be crucial to the distribution of sessile organisms in marine environments.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Lectins probe molecular films in biofouling:characterization of early films on non-living and living surfaces

Michael¹,

Smith²

1995

Mar. Ecol. Prog. Ser.

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“…Lectins conjugated to fluorescent dyes have been used to detect carbohydrate residues in mucilage and cell walls of microalgae (von Sengbusch et al, 1982;von Sengbusch & Müller, 1983;Callow, 1985).…”

Section: Introductionmentioning

confidence: 99%

Spore Adhesion and Cell Wall Formation in Gelidium Floridanum (Rhodophyta, Gelidiales)

2006

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The attachment of spores to a substratum is essential for their germination and, therefore, to the completion of the life cycle of the red algae. In most red algae, spores are liberated without a cell wall, within a sheath of mucilage which is responsible for their primary attachment. Utilizing fluorescent-labeled lectins, we identified carbohydrate residues and their locations in the mucilage and cell walls of spores of Gelidium floridanum. Cell wall formation and mucilage composition were studied with calcofluor, toluidine blue -O (AT-O), alcian blue (AB) and periodic acid-Schiff (PAS). In the mucilage we identified α-D mannose, α-D glucose, β-D-galactose, N-acetyl-glucosamine and N-acetyl-galactosamine. The first two sugar residues were not found in the cell wall of the germ tube but they were present on the rhizoid's cell wall indicating their importance to substrate adhesion. A cell wall is produced soon after the spore's attachment, beginning with a polar deposition of cellulose and its gradual spread around the spore as indicated by calcofluor. The cell wall matrix was positive to AB and metachromatic to AT-O, indicating acidic polysaccharides, while cellulose microfibrills were positive to PAS. A polar disorganization of the cell wall triggers the process of germination. As spores are the natural form of propagation of Gelidium, the understanding of the mechanisms of spore attachment may contribute to the cultivation of this valuable seaweed.

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“…All parameters can be assessed on the species level or even on level of sub-populations (Reckermann and Veldhuis, 1997). were mainly used to study class and cell structure specific binding (Sengbusch et al, 1982;Sengbusch and Müller, 1983;Costas and Rodas, 1994;Costas et al, 1995). Alternative stains to detect specific cellular compounds are Calcofluor White-ST, which stains typically cellulose and chitin fibres, the lipid stains Neutral Red (Crippen, 1974) and Nile Red (Cooksey et al, 1987;Macho et al, 1996).…”

Section: Growth Ratementioning

confidence: 99%

Application of flow cytometry in marine phytoplankton research: current applications and future perspectives

Veldhuis¹,

Kraay²

2000

Sci. Mar.

169

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SUMMARY: A brief overview is given of current applications of flow cytometry (FCM) in marine phytoplankton research. This paper presents a selection of highlights and various technical and analytical problems we encountered during the past 10 years. In particular, the conversion of the relative values obtained in terms of size and fluorescence applying FCM to quantitative estimates of cell size, pigment concentration, genome size etc., is addressed. The introduction of DNA -cellcycle analysis made easily assessable by flow cytometry has been of great importance, allowing in situ measurement of species specific growth rates. Key questions in ecology such as factors determining the wax and wane of phytoplankton bloom can now be better answered in terms of species specific growth and mortality. Finally, flow cytometry provides detailed information of the physiological status of the individual algal cells. New staining methods enable us to distinguish between viable and non-viable cells and so will help us to elucidate the importance of "automortality" in aquatic ecosystems.

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FITC-labeled lectins and calcofluor white ST as probes for the investigation of the molecular architecture of cell surfaces. Studies on conjugatophycean species

Cited by 31 publications

References 33 publications

Lectins probe molecular films in biofouling:characterization of early films on non-living and living surfaces

Lectins probe molecular films in biofouling:characterization of early films on non-living and living surfaces

Spore Adhesion and Cell Wall Formation in Gelidium Floridanum (Rhodophyta, Gelidiales)

Application of flow cytometry in marine phytoplankton research: current applications and future perspectives

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