Direct covalent linkage of fluorescent probes to the plant protoplast surface

Furtula, Vesna; Walko, R. M.; Nothnagel, Eugene A.

doi:10.1007/bf01282282

Cited by 10 publications

(4 citation statements)

References 31 publications

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“…The tegument in helminths and nematodes is rich in carbohydrates and lipids [29] , [30] . Therefore, using two methods the organism was labeled with hydrazide groups that are known to bind oxidizable aldehyde groups which are highly represented in molecules containing carbohydrates and lipids [31] . M. corti metacestodes labeled using the periodate method showed strong staining in the tegument's distal cytoplasm ( Figure 2A–2B ) but not in other areas.…”

Section: Resultsmentioning

confidence: 99%

Differential Release and Phagocytosis of Tegument Glycoconjugates in Neurocysticercosis: Implications for Immune Evasion Strategies

2008

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Neurocysticercosis (NCC) is an infection of the central nervous system (CNS) by the metacestode of the helminth Taenia solium. The severity of the symptoms is associated with the intensity of the immune response. First, there is a long asymptomatic period where host immunity seems incapable of resolving the infection, followed by a chronic hypersensitivity reaction. Since little is known about the initial response to this infection, a murine model using the cestode Mesocestoides corti (syn. Mesocestoides vogae) was employed to analyze morphological changes in the parasite early in the infection. It was found that M. corti material is released from the tegument making close contact with the nervous tissue. These results were confirmed by infecting murine CNS with ex vivo–labeled parasites. Because more than 95% of NCC patients exhibit humoral responses against carbohydrate-based antigens, and the tegument is known to be rich in glycoconjugates (GCs), the expression of these types of molecules was analyzed in human, porcine, and murine NCC specimens. To determine the GCs present in the tegument, fluorochrome-labeled hydrazides as well as fluorochrome-labeled lectins with specificity to different carbohydrates were used. All the lectins utilized labeled the tegument. GCs bound by isolectinB4 were shed in the first days of infection and not resynthesized by the parasite, whereas GCs bound by wheat germ agglutinin and concavalinA were continuously released throughout the infectious process. GCs bound by these three lectins were taken up by host cells. Peanut lectin-binding GCs, in contrast, remained on the parasite and were not detected in host cells. The parasitic origin of the lectin-binding GCs found in host cells was confirmed using antibodies against T. solium and M. corti. We propose that both the rapid and persistent release of tegumental GCs plays a key role in the well-known immunomodulatory effects of helminths, including immune evasion and life-long inflammatory sequelae seen in many NCC patients.

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Section: Resultsmentioning

confidence: 99%

Differential Release and Phagocytosis of Tegument Glycoconjugates in Neurocysticercosis: Implications for Immune Evasion Strategies

2008

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“…The lipid component of the protoplast plasma membrane was fluorescence-labeled with Lucifer yellow-cholesterol (LY-Chol) as described by Walko and Nothnagel (1989). Plasma-membrane proteins and glycoconjugates were fluorescence-labeled with Lucifer yellow-vinyl-sulfone (LY-VS) and fluorescein thiosemicarbazide (FTSC), respectively, following the procedure described by Furtula et al (1987). The FTSC was attached at aldehydes generated by periodate cleavage and has been shown to label AGPs as well as other plasma-membrane glycoconjugates .…”

Section: Measurements Of Lateral Diffusion Of Plasma-membrane Componementioning

confidence: 99%

Effects of Yariv phenylglycosides onRosa cell suspensions: Evidence for the involvement of arabinogalactan-proteins in cell proliferation

Serpe

Nothnagel

1994

Planta

171

161

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“…Published micrographs showing predominant fluorescence at the protoplast surface are available for maize root cortex protoplasts labelled with either LY-DC,2:oPE (Furtula et ai 1990) or LY-Chol (Nothnagel 1989), a cholesterol probe with structural and labelling characteristics nearly identical to those of LY-Sito. A comparative study reporting the persistence of LY-VS labelling at the maize root protoplast surface is also available (Furtula et at. 1987).…”

Section: Discussionmentioning

confidence: 99%

“…Protoplasts were labelled either with 400 g m~^ LY-DCi2:oPE or with 100 g m"^ LY-Sito for 20-30 min at 26 °C, and then excess probe was removed by centrifugation in buffered mannitol solution at 200 g two times for 6 min each. Proteins in the plasma membrane were labelled by incubation of protoplasts with LY-VS (Lucifer Yellow VS), as described by Furtula, Walko & Nothnagel (1987).…”

Section: Fluorescence Labelling and Fpr Measurementsmentioning

confidence: 99%

Effects of low temperature on lateral diffusion in plasma membranes on maize (Zea mays L.) root cortex protoplasts: relevance to chilling sensitivity

Koster

Tengbe

Furtula

et al. 1994

Plant Cell & Environment

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The effects of temperature on the lateral diffusion of fluorescent phospholipids, sterols and proteins in the plasma membranes of maize root cortex protoplasts were monitored using fluorescence photobleaching recovery (FPR). Diffusion parameters were measured in two cultivars of maize having different chilling tolerance. Hydrodynamic theory predicts that the diffusion coefficient, D, should increase with increasing temperature. In the more chilling-tolerant cultivar, however, D for all three prohes was nearly insensitive to temperature. In the more chillingsensitive cultivar, D was also insensitive to temperature over the range from 12 to 21 °C, but D for the lipid probes tended to be higher and more variable at lower temperatures. The proportion of probe molecules free to diffuse in the membrane was less than 1 for all probes, and increased significantly with increasing temperature for the protein probe. These results, taken together, support the concept that the plasma membrane contains domains having differing diffusional characteristics. Temperature effects on membrane diffusion are moderated by the existence of these domains to limit significant changes. The observed tendency for higher diffusion coefficients at low temperatures in the chilling-sensitive cultivar may correlate to morphological changes observed with protoplasts of that cultivar at low temperatures.

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Direct covalent linkage of fluorescent probes to the plant protoplast surface

Cited by 10 publications

References 31 publications

Differential Release and Phagocytosis of Tegument Glycoconjugates in Neurocysticercosis: Implications for Immune Evasion Strategies

Differential Release and Phagocytosis of Tegument Glycoconjugates in Neurocysticercosis: Implications for Immune Evasion Strategies

Effects of Yariv phenylglycosides onRosa cell suspensions: Evidence for the involvement of arabinogalactan-proteins in cell proliferation

Effects of low temperature on lateral diffusion in plasma membranes on maize (Zea mays L.) root cortex protoplasts: relevance to chilling sensitivity

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