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Subcellular Fractionation of Rat Sciatic Nerve and Specific Localization of Ganglioside LM 1 in Rat Nerve Myelin
Author(s) -
Chou K. H.,
Nolan C. E.,
Jungalwala F. B.
Publication year - 1985
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1985.tb07186.x
Subject(s) - myelin , ganglioside , sciatic nerve , peripheral nervous system , chemistry , microsome , biochemistry , central nervous system , cell fractionation , membrane , differential centrifugation , nervous system , biology , anatomy , enzyme , endocrinology , neuroscience
Subcellular fractionation of rat sciatic nerve was developed to determine the specific localization of gangliosides in the nerve membrane fractions. Myelin, microsomal, and a plasma membrane‐like fraction were isolated and purified by sucrose density gradient centrifugation. These subfractions were characterized by electron microscopy, marker enzyme assays, and their protein and lipid profile. In rat sciatic nerve myelin, 90 mol% of the total gangliosides were monosialogangliosides. LM 1 (sialosyl‐lactoneotetraosylceramide) (61 mol%) and GM 3 (21%) were the major gangliosides of the rat nerve myelin. Two other neolacto series of gangliosides, viz., sialosyl‐lactoneonorhexaosylceramide and sialosyl‐lactoneooctasylceramide, were also localized mostly in the myelin fraction. GM 1 was only a minor (< 2%) ganglioside in myelin. The ganglioside patterns of the microsomal and plasma membrane‐like fractions were similar with minor quantitative differences and were entirely different from that of myelin. Monosialogangliosides were –70–75 mol% of the total in these fractions. The major gangliosides of the microsomal and plasma membrane‐like fractions were GM 3 (–40%) and GM 1 (–20%). LM 1 in these fractions was minimal (less than –5%). Significant amounts of GM 3 with N ‐glycolylneuraminic acid (–10%) and GM 1b (4–14%) were also identified in the microsomal and plasma membrane‐like fractions but not in myelin. These and the higher lactoneo series of gangliosides have not been previously reported to be present in the rat nervous system. Almost exclusive localization of LM 1 in myelin in rat peripheral nervous system is consistent with our previous observation that deposition of LM 1 in the nerve with age was very similar to that of myelin marker lipids cerebrosides and sulfatides.