Complex glycosphingolipids of the pheochromocytoma cell line PC12: Enhanced fucosylglycolipid synthesis following nerve growth factor treatment

Schwarting, Gerald A.; Gajewski, Anna; Barbero, Linda; Tischler, Arthur S.; Costopoulos, Donna

doi:10.1016/0306-4522(86)90287-3

Cited by 20 publications

(11 citation statements)

References 25 publications

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“…Schwarting et al [7] reported that fucoglycolipid synthesis was increased by about 50% following NGF treatment of PC12 cells in spinner cell cultures. Interest ingly, fucosylated neutral glycolipids were increased whereas non fucosylated neutral glycolipids were not, suggesting that the reg ulation of fucosylation of neutral glycolipids was distinct from the bulk of glycolipid glycosylation in PC12 cells.…”

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confidence: 99%

Fucosylation of Glycolipids in PC12 Cells Is Dependent on the Sequence of Nerve Growth Factor Treatment and Adenylate Cyclase Activation

1990

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Synthesis of complex glycosphingolipids was analyzed in cultures of PC 12 cells treated with nerve growth factor (NGF) and forskolin, either sequentially or simultaneously. For the sequential treatment, forskolin was added following 15 days of NGF treatment and cultures were continued for 3 additional days. For simultaneous treatment, cells maintained in medium with no additives received both agents and were then incubated for 3 additional days. Sequential NGF/forskolin treatment induced hypertrophy in PC12 cells and produced cells which had more neuron-like characteristics than cells treated with NGF alone. Simultaneous treatment initially accelerated the outgrowth of neurites and then reduced neurite elongation. Sequential treatment of PC12 cells resulted in a greater than 50% increase in the incorporation of fucose into neutral glycolipids and gangliosides compared to treatment with NGF alone. Galactose incorporation was unchanged, indicating that there was no net increase in glycolipid synthesis. The lack of an increase in total glycolipid accumulation was confirmed using monoclonal antibodies reacting with specific fucosylated and nonfucosylated PC12 cell glycolipids. However, TLC immunostaining analysis revealed that sequential NGF/forskolin treatment selectively altered the expression of some neutral glycolipid species. In contrast to the effects of the sequential treatment, simultaneous NGF and forskolin treatment was accompanied by a decrease in the incorporation of fucose and galactose into neutral glycolipids and gangliosides. These findings suggest that the timing of exposure to neuronotrophic factors and agents which activate adenylate cyclase might in part account for developmental regulated patterns of glycoconjugate expression and provide further evidence that fucosylation of glycolipids may be associated with maturation of sympathetic neurons.

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confidence: 99%

Fucosylation of Glycolipids in PC12 Cells Is Dependent on the Sequence of Nerve Growth Factor Treatment and Adenylate Cyclase Activation

1990

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“…Some of these species may be the other metabolites shown in Figure 1A not previously detected. These peaks may also be other GSL species; dPC12 cells are known to express even more complex fucose-containing GSLs [58–60]. New fluorescent GSL standards will need to be synthesized [32] to identify these unknown peaks.…”

Section: Resultsmentioning

confidence: 99%

Manipulating ionic strength to improve single cell electrophoretic separations

Keithley

Metzinger

Rosado

et al. 2013

Talanta

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A capillary electrophoresis system with ultrasensitive two-color laser-induced fluorescence detection was used to probe the effect of ionic strength on single cell separations of glycosphingolipids. Differentiated PC12 cells were incubated with two ganglioside substrates tagged with different fluorophores within the BODIPY family such that two distinct metabolic patterns could be simultaneously monitored. Aspiration of single differentiated PC12 cells suspended in a phosphate-buffered saline solution showed excessive peak dispersion, poor resolution, and peak efficiencies below 100,000 theoretical plates. Aspiration of single differentiated PC12 cells suspended in deionized water corrected peak dispersion. Average peak efficiencies ranged between 400,000 and 600,000 theoretical plates. Improved performance was due to the dilution of the high salt concentrations inside of single neuronal-like cells to produce field amplified sample stacking. Single cell separations showed the highest resolution when aspiration of single differentiated PC12 cells suspended in deionized water were separated using a running buffer of high ionic strength. The improvement in resolution allowed for the identification of analytes not previously detected in single cell metabolism studies.

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“…The role of fucose in the GSLs of vertebrates has yet to be fully understood. It has been shown that nerve growth factor treatment of the pheochromocytoma cell line PC 12 enhanced fucosylglycolipid synthesis [38] and polyunsaturated, fucosylated GSLs have been found to be essential [39] for spermatogenesis and male mouse fertility. A fucosylated glycosphingolipid has been isolated from the millipede, Parafontaria laminate [40], and the structure was characterized as Manβ1-4(Fucα-3)Glcβ1-ceramide.…”

Section: Discussionmentioning

confidence: 99%

Structural studies on the neutral glycosphingolipids of Manduca sexta

Abeytunga

Oland

Somogyi

et al. 2008

Bioorganic Chemistry

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Glycosphingolipids (GSLs) have been implicated as playing major roles in cellular interactions and control of cell proliferation in muticellular organisms. Moreover GSLs and other sphingolipids such as sphingomyelins, ceramides and sphingosines serve a variety of roles in signal transduction. Hence, identification of structures of GSLs in different biota will shed light in understanding their physiological role. During this study, the major glycosphingolipid component present in the extracts of stage-12 and stage-17/18 metamorphosing adults of Manduca sexta was identified as mactosyl ceramide. We report the isolation of several ceramide disaccharides, a ceramide trisaccharide and a ceramide tetrasaccharide. The GSL structures were confirmed by high-resolution mass spectrometry and tandem mass spectrometry. The identity of the monosaccharides was proved using exoglycosidases. The predominant sphingosine chain-length varied from C-14 (tetradecasphing-4-enine) to C-16 (hexadecasphing-4-enine) in these GSLs. Sphingosines of both chain lengths were accompanied by their doubly unsaturated counterparts tetradecasphinga-4,6-diene and hexadecasphinga-4,6-diene. It is also interesting to note the presence of tetradecasphinganine and hexadecasphinganine in minute amounts in the form of a GSL in the extracts of M. sexta. The varying degrees of unsaturation in the sphingosine moiety of GSLs in M. sexta may be biologically significant in insect metamorphosis. The ceramide trisaccharides and ceramide tetrasaccharide belong to the arthro-series, The observation of fucose in the M. sexta GSLs is the first report of the presence of fucose in an arthroseries GSL.

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Complex glycosphingolipids of the pheochromocytoma cell line PC12: Enhanced fucosylglycolipid synthesis following nerve growth factor treatment

Cited by 20 publications

References 25 publications

Fucosylation of Glycolipids in PC12 Cells Is Dependent on the Sequence of Nerve Growth Factor Treatment and Adenylate Cyclase Activation

Fucosylation of Glycolipids in PC12 Cells Is Dependent on the Sequence of Nerve Growth Factor Treatment and Adenylate Cyclase Activation

Manipulating ionic strength to improve single cell electrophoretic separations

Structural studies on the neutral glycosphingolipids of Manduca sexta

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