Synthesis and mass spectrometric characterization of digoxigenin and biotin labeled ganglioside GM I and their uptake by and metabolism in cultured cells

Albrecht, Bernd; Pohlentz, Gottfried; Sandhoff, Konrad; Schwarzmann, Günter

doi:10.1016/s0009-3084(97)02658-3

Cited by 15 publications

(13 citation statements)

References 35 publications

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“…Dietary sulfatide, an acidic glycosphingolipid, has been shown to be absorbed in rat ileum with small amounts incorporated in 2 h, larger amounts incorporated in 6 h, and most sulfatide disappearing from the intestine by 24 h (7). When present for 24 -72 h at 37°C in culture, GM1 is incorporated into fibroblasts in the range of a few nanomoles per milligram cellular protein (1,34). Compared with glycolipid uptake studies using other cell lines, the time frame for GD3 uptake by intestinal cells in the present study was similar.…”

Section: Discussionsupporting

confidence: 53%

“…In contrast to GM1, GD3 was incorporated in the range of a few micrograms per mg cell protein. Given that the amount of GM1 given to cells was similar or higher (1-150 g/ml) than the amount of GD3 used in this absorption study (5-10 g/ml), the present study may suggest that GD3 is more bioavailable than GM1 (1,34,46,53).…”

Section: Discussionmentioning

confidence: 67%

“…Caco-2 cells were incubated under standard conditions in the absence (control, MEM-4 alone) or presence of purified ganglioside GD3 in MEM-4 for 6, 24, and 48 h at a concentration of 5 or 10 g/ml. The time frame for exposing human intestine Caco-2 cells to ganglioside GD3 was chosen to be 6 -48 h because previous work on ganglioside GM1 uptake by cultured cells predicts slow and incomplete ganglioside absorption from the apical side (1,34). The GD3 concentration chosen for absorption experiments is physiological: close to the concentration of gangliosides in mature human milk (9.07 g sialic acid per milliliter) and high enough to exert biological effects without exhibiting toxicity (8,59).…”

Section: Methodsmentioning

confidence: 99%

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Uptake and fate of ganglioside GD3 in human intestinal Caco-2 cells

Schnabl¹,

Larcelet²,

Thomson³

et al. 2009

American Journal of Physiology-Gastrointestinal and Liver Physi

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Ganglioside GD3 is a glycosphingolipid found in colostrum, developing tissues, and tumors and is known to regulate cell growth, differentiation, apoptosis, and inflammation. Feeding a GD3-enriched diet to rats increases GD3 in intestinal lipid rafts and blood. The mechanism, efficiency, and fate of ganglioside absorption by human enterocytes have not been investigated. A model to study GD3 uptake by human intestinal cells was developed to test the hypothesis that enterocyte GD3 uptake is time and concentration dependent, with uptake efficiency and fate influenced by route of delivery. Caco-2 cells were exposed to GD3 on the apical or basolateral membrane (BLM) side for 6, 24, and 48 h. GD3 uptake, retention, transfer, and metabolism was determined. GD3 uptake across the apical and BLM was time and concentration dependent and reached a plateau. GD3 uptake across the BLM was more efficient than apical delivery. Apical GD3 was metabolized with some cell retention and transfer, whereas basolateral GD3 was mostly metabolized. This study demonstrates efficient GD3 uptake by enterocytes and suggests that the route of delivery influences ganglioside uptake and fate.

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Section: Discussionsupporting

confidence: 53%

Section: Discussionmentioning

confidence: 67%

Section: Methodsmentioning

confidence: 99%

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Uptake and fate of ganglioside GD3 in human intestinal Caco-2 cells

Schnabl¹,

Larcelet²,

Thomson³

et al. 2009

American Journal of Physiology-Gastrointestinal and Liver Physi

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“…In principle, the binding of gangliosides to plasma membranes occurs in three different GM1 regulates APP proteolysis Q Zha et al operationally determined manner: loosely associated micelles removable by serum proteins, a proteinbound, serum-resistant but trypsin-labile ganglioside fraction, and trypsin-stable associated gangliosides largely comprising inserted monomers. 31,32 The insertion into plasma membranes of gangliosides from their micellar state is a slow process. The maximal incorporation into cells of GM1 was reached after 72 h. The exogenous GM1 that has been incorporated into the outer surface of the plasma membrane becomes internalized.…”

Section: Discussionmentioning

confidence: 99%

GM1 ganglioside regulates the proteolysis of amyloid precursor protein

et al. 2004

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Plaques containing amyloid b-peptides (Ab) are a major feature in Alzheimer's disease (AD), and GM1 ganglioside is an important component of cellular plasma membranes and especially enriched in lipid raft. GM1-bound Ab (GM1/Ab), found in brains exhibiting early pathological changes of AD including diffuse plaques, has been suggested to be involved in the initiation of amyloid fibril formation in vivo by acting as a seed. However, the role of GM1 in amyloid bprotein precursor (APP) processing is not yet defined. In this study, we report that exogenous GM1 ganglioside promotes Ab biogenesis and decreases sAPPa secretion in SH-SY5Y and COS7 cells stably transfected with human APP695 cDNA without affecting full-length APP and the sAPPb levels. We also observe that GM1 increases extracellular levels of Ab in primary cultures of mixed rat cortical neurons transiently transfected with human APP695 cDNA. These findings suggest a regulatory role for GM1 in APP processing pathways. Molecular Psychiatry (2004) 9, 946-952.

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“…For this purpose we synthesized a biotin-tagged GM1 derivative (biotin-GM1; Albrecht et al 1997) to study its intracellular transport along the endocytic pathway by electron microscopy using goldconjugated anti-biotin antibodies. Another important prerequisite for visualization is the resistance of the biotin label to degradation by cells.…”

mentioning

confidence: 99%

Intracellular Distribution of a Biotin-labeled Ganglioside, GM1, by Immunoelectron Microscopy After Endocytosis in Fibroblasts

Möbius

Herzog

Sandhoff

et al. 1999

J Histochem Cytochem.

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A radioactive and biotin-labeled analogue of GM1 (biotin-GM1) was synthesized which enabled us to analyze its intracellular distribution in the compartments of the endocytic route by electron microscopic immunocytochemistry using thin sections of human skin fibroblasts labeled with gold-conjugated antibiotin antibodies. Metabolic studies with the biotin-GM1 showed its partial degradation to the corresponding GM2 and GM3 derivatives. Further degradation was inhibited by the biotin residue. The distribution of biotin-GM1 after uptake by cells was studied by postembedding labeling techniques. On the plasma membrane the biotin-GM1 was detectable in the form of patches (0.1 micrometer in diameter), in caveola-like structures and, to a much lesser extent, in coated pits or vesicles. During endocytic uptake, the biotin-GM1 became detectable in organelles identified as late endosomes and lysosomes. The intracellular distribution of the biotin-GM1 was compared to the localization of the EGF receptor in EGF-stimulated fibroblasts. Both the biotin-GM1 and the EGF receptor were transported to intraendosomal and intralysosomal membranes, indicating that both membrane constituents follow the same pathway of endocytosis. Our observations show that biotin-GM1 can be successfully incorporated into the plasma membrane and be used as a tool for morphological detection of its pathway to lysosomes.

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Synthesis and mass spectrometric characterization of digoxigenin and biotin labeled ganglioside GM I and their uptake by and metabolism in cultured cells

Cited by 15 publications

References 35 publications

Uptake and fate of ganglioside GD3 in human intestinal Caco-2 cells

Uptake and fate of ganglioside GD3 in human intestinal Caco-2 cells

GM1 ganglioside regulates the proteolysis of amyloid precursor protein

Intracellular Distribution of a Biotin-labeled Ganglioside, GM1, by Immunoelectron Microscopy After Endocytosis in Fibroblasts

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