Ganglioside-Induced Neuritogenesis: Verification That Gangliosides Are the Active Agents, and Comparison of Molecular Species |
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Authors: | Mary C Byrne Robert W Ledeen Fred J Roisen † Glee Yorke † Joseph R Sclafani |
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Institution: | Department of Neurology and Biochemistry, Albert Einstein College of Medicine, Bronx, New York;Department of Anatomy, University of Medicine and Dentistry of New Jersey-Rutgers Medical School, Piscataway, New Jersey, U.S.A. |
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Abstract: | Abstract: Gangliosides were previously reported to induce neuritogenesis in primary neuronal cultures and in some neurally derived cell lines. Because isolated gangliosides usually contain variable quantities of peptides, we investigated the possibility the neurite-stimulating activity could be caused by these contaminants. Ganglioside preparations from bovine brain and other sources were subjected to a three-step purification procedure that eliminated at least 95% of the contaminating peptides. These purfied preparations retained their capacity to induce extensive neurite growth in neuro-2A murine neuroblastoma. Proteolytic digestion and a number of additional procedures were used to reduce residual contamination further without loss of activity. Several crude ganglioside samples had negative effects on neurite development until freed of theri inhibitory factors, which were derived from the tissue and/or introduced during laboratory operations. This was particularly evident for bovine white matter gangliosides whose activity increased in proportion to peptide removal. When carefully purified, virtually all of 11 different gangliosides tested were highly active, with the possible exception of GM4, which demonstrated only moderate activity in a limited number of tests. All of the neutral glycolipids tested, as well as sulfatides and free sialic acid, were inactive. |
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Keywords: | Ganglioside Neuritogenesis N-2A cells Ganglioside purification Peptide contaminants |
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