A New Type of Two-Dimensional Thin-Layer Chromatography (TLC Mapping) for Analysis of Acidic Glycosphingolipid Molecular Species

Watanabe, K.; Nishiyama, M

doi:10.1006/abio.1995.1270

Cited by 9 publications

(5 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Interestingly, ethanol dehydration markedly reduced overall levels of renal Gb 3 but only minimally decreased levels of neuronal Gb 3 . This result may be explained partly by the distinct glycolipid proWles of nervous and renal tissue (Penick et al 1966;Rauvala 1976;Watanabe and Nishiyama 1995); such diVerences in lipid composition may eVect the membrane environment and alter lipid solubility and extraction eYciency (Christie 1993).…”

Section: Discussionmentioning

confidence: 95%

Immunohistologic techniques for detecting the glycolipid Gb3 in the mouse kidney and nervous system

Kolling

Obata

Gross

et al. 2008

Histochem Cell Biol

View full text Add to dashboard Cite

Shiga toxin-producing Escherichia coli causes hemolytic uremic syndrome, a constellation of disorders that includes kidney failure and central nervous system dysfunction. Shiga toxin binds the amphipathic, membrane-bound glycolipid globotriaosylceramide (Gb(3)) and uses it to enter host cells and ultimately cause cell death. Thus, cell types that express Gb(3) in target tissues should be recognized. The objective of this study was to determine whether immunohistologic detection of Gb(3) was affected by the method of tissue preparation. Tissue preparation included variations in fixation (immersion or perfusion) and processing (paraffin or frozen) steps; paraffin processing employed different dehydration solvents (acetone or ethanol). Perfusion-fixation in combination with frozen sections or acetone-dehydrated tissue for paraffin sections resulted in specific recognition of Gb(3) using immunohistochemical or immunofluorescent methods. In the mouse tissues studied, Gb(3) was associated with tubules in the kidney and neurons in the nervous system. On the other hand, Gb(3) localization to endothelial cells was determined to be an artifact generated due to immersion-fixation or tissue dehydration with ethanol. This finding was corroborated by glycolipid profiles from tissue subjected to dehydration; namely Gb(3) was subject to extraction by ethanol more than acetone during tissue dehydration. The results of this study show that tissue preparation is crucial to the persistence and preservation of the glycolipid Gb(3) in mouse tissue. These methods may serve as a basis for determining the localization of other amphipathic glycolipids in tissue.

show abstract

Section: Discussionmentioning

confidence: 95%

Immunohistologic techniques for detecting the glycolipid Gb3 in the mouse kidney and nervous system

Kolling

Obata

Gross

et al. 2008

Histochem Cell Biol

View full text Add to dashboard Cite

show abstract

“…If separation is insufficient with conventional single run TLC, multiple development of the plate can be carried out in three ways: in the original direction of development with (1) the same solvent, (2) different solvents, or (3) at right angles to the original direction of the chromatography and with a different solvent (¼two-dimensional TLC). All three alternatives increase the resolution capability of TLC by affording separations not otherwise attainable (Rösner, 1980;Nakamura et al, 1983;Sonnino et al, 1983;Yao & Rastetter, 1985;Watanabe & Nishiyama, 1995;Yohe et al, 2001). An improved separation of gangliosides can also be achieved by unidirectional automated multiple separation performing, for example, three consecutive runs, each with intermediate vacuum drying periods, in the same solvent but with lower polarity than that which is optimal for separation by conventional single TLC (Müthing, 1994;Müthing & Ziehr, 1996).…”

Section: A Basicsmentioning

confidence: 97%

Advances on the compositional analysis of glycosphingolipids combining thin-layer chromatography with mass spectrometry

Müthing

Distler

2009

Mass Spectrom. Rev.

View full text Add to dashboard Cite

Glycosphingolipids (GSLs), composed of a hydrophilic carbohydrate chain and a lipophilic ceramide anchor, play pivotal roles in countless biological processes, including infectious diseases and the development of cancer. Knowledge of the number and sequence of monosaccharides and their anomeric configuration and linkage type, which make up the principal items of the glyco code of biologically active carbohydrate chains, is essential for exploring the function of GSLs. As part of the investigation of the vertebrate glycome, GSL analysis is undergoing rapid expansion owing to the application of novel biochemical and biophysical technologies. Mass spectrometry (MS) takes part in the network of collaborations to further unravel structural and functional aspects within the fascinating world of GSLs with the ultimate aim to better define their role in human health and disease. However, a single-method analytical MS technique without supporting tools is limited yielding only partial structural information. Because of its superior resolving power, robustness, and easy handling, high-performance thin-layer chromatography (TLC) is widely used as an invaluable tool in GSL analysis. The intention of this review is to give an insight into current advances obtained by coupling supplementary techniques such as TLC and mass spectrometry. A retrospective view of the development of this concept and the recent improvements by merging (1) TLC separation of GSLs, (2) their detection with oligosaccharide-specific proteins, and (3) in situ MS analysis of protein-detected GSLs directly on the TLC plate, are provided. The procedure works on a nanogram scale and was successfully applied to the identification of cancer-associated GSLs in several types of human tumors. The combination of these two supplementary techniques opens new doors by delivering specific structural information of trace quantities of GSLs with only limited investment in sample preparation.

show abstract

“…Minor, unique gangliosides, 7-0-acetyl, 9-0-acetyl, and 7,9-di-0-acetyl GD , have been found in buttermilk (Huang, 1973;Bonafede et al, 1989;Ren et al, 1992) and recently in bovine cheese whey and human milk (Hanagata et al, 1991;Watanabe & Nishiyama, 1995) suggesting cultured dairy products may show other ganglioside variations.…”

Section: Discussionmentioning

confidence: 95%

Variation in Ganglioside Content of Bovine Dairy Products

Moore

Ettinger

Yokoyama

2000

Journal of Food Composition and Analysis

View full text Add to dashboard Cite

A New Type of Two-Dimensional Thin-Layer Chromatography (TLC Mapping) for Analysis of Acidic Glycosphingolipid Molecular Species

Cited by 9 publications

References 0 publications

Immunohistologic techniques for detecting the glycolipid Gb3 in the mouse kidney and nervous system

Immunohistologic techniques for detecting the glycolipid Gb3 in the mouse kidney and nervous system

Advances on the compositional analysis of glycosphingolipids combining thin-layer chromatography with mass spectrometry

Variation in Ganglioside Content of Bovine Dairy Products

Contact Info

Product

Resources

About