Regulation of apoptosis during neuronal differentiation by ceramide and b-series complex gangliosides.

Abstract

Lipid analysis of gestational day E14.5 mouse brain revealed elevation of ceramide to a tissue concentration that induced apoptosis when added to the medium of neuroprogenitor cells grown in cell culture. Elevation of ceramide was coincident with the first appearance of b-series complex gangliosides (BCGs). Expression of BCGs by stable transfection of murine neuroblastoma (F-11) cells with sialyltransferase-II (ST2) resulted in a 70% reduction of ceramide-induced apoptosis. This was most likely due to an 80% reduced expression of prostate apoptosis response-4 (PAR-4). PAR-4 expression and apoptosis were restored by preincubation of ST2-transfected cells with N-butyl deoxinojirimycin (NB-DNJ) or PD98059, two inhibitors of ganglioside biosynthesis or p42/44 mitogen-activated protein (MAPK) kinase, respectively. In sections of day E14.5 mouse brain, the intermediate zone showed intensive staining for complex gangliosides, but only low staining for apoptosis (TUNEL) and PAR-4. Apoptosis and PAR-4 expression, however, were elevated in the ventricular zone which only weakly stained for complex gangliosides. Whole cell patch clamping revealed a 2-fold increased calcium influx in ST2-transfected cells, the blocking of which with nifedipine restored apoptosis to the level of untransfected cells. In serum-free culture, supplementation of the medium with IGF-1 was required to maintain MAPK phosphorylation and the anti-apoptotic effect of BCG expression. BCG-enhanced calcium influx and the presence of insulin-like growth factor-1 may thus activate a cell survival mechanism that selectively protects developing neurons against ceramide-induced apoptosis by up-regulation of MAPK and reduction of PAR-4 expression.

• Authors: E Bieberich, S MacKinnon, J Silva, R K Yu
• Journal: The Journal of biological chemistry (J Biol Chem) Vol. 276 Issue 48 Pg. 44396-404 (Nov 30 2001) ISSN: 0021-9258 [Print] United States
• PMID: 11574545 (Publication Type: Journal Article, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Apoptosis Regulatory Proteins
• Carrier Proteins
• Ceramides
• Enzyme Inhibitors
• Flavonoids
• Gangliosides
• Intracellular Signaling Peptides and Proteins
• Luminescent Proteins
• Recombinant Fusion Proteins
• prostate apoptosis response-4 protein
• Green Fluorescent Proteins
• 1-Deoxynojirimycin
• Mitogen-Activated Protein Kinase 1
• 2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one

Topics

• 1-Deoxynojirimycin (pharmacology)
• Animals
• Apoptosis
• Apoptosis Regulatory Proteins
• Brain (embryology, pathology)
• Carrier Proteins (biosynthesis)
• Cell Differentiation
• Cells, Cultured
• Ceramides (pharmacology)
• Electrophoresis, Polyacrylamide Gel
• Enzyme Inhibitors (pharmacology)
• Flavonoids (pharmacology)
• Gangliosides (metabolism)
• Green Fluorescent Proteins
• In Situ Nick-End Labeling
• Intracellular Signaling Peptides and Proteins
• Luminescent Proteins (metabolism)
• Membrane Potentials
• Mice
• Mitogen-Activated Protein Kinase 1 (metabolism)
• Neuroblastoma (metabolism)
• Neurons (cytology)
• Patch-Clamp Techniques
• Phosphorylation
• Recombinant Fusion Proteins (metabolism)
• Signal Transduction
• Time Factors
• Tissue Distribution
• Transfection
• Tumor Cells, Cultured
• Up-Regulation