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J. Biochem, 1977, Vol. 81, No. 5 1217-1225
© 1977 Japanese Biochemical Society

research-article

A New Procedure for the Isolation of Brain Gangliosides, and Determination of Their Long Chain Base Compositions

Nariko KAWAMURA and Tamotsu TAKETOMI

Department of Biochemistry, Institute of Adaptation Medicine, Shinshu University Matsumoto, Nagano 390

Total gangliosides were isolated from grey and white matter of animal brains and human brains at various ages, including the pathological brain of a patient with late infantile amaurotic family idiocy (Bielschowsky-Jansky disease) by a new procedure. Total lipids extracted from brain tissues were directly applied to a colunm of Silica Gel G-Silica Guhr G (1:1, w/w) slurried with chloroform-methanol-28% ammonia (32:8:1, by vol) without Folch's partition. After the total lipids, with the exception of gangliosides, had been eluted with the same solvent, the total gangliosides were eluted with chloroform-methanol-water (60:35:8, by vol). The total gangliosides were subjected to mild alkaline methanolysis and dialyzed against deionized water, then the ganglioside fraction thus obtained was rechromatographed in the same way. The content of gangliosides in various brains was 400–500 µg for grey matter and 150–200 µg for white matter (amount of long chain base per gram of fresh tissue). GD1a and GM1 accounted for about 30–40% of the total gangliosides in grey matter, while in white matter GM1 was the major ganglioside (>50%): it was particularly noteworthy that sialosylgalactosyl ceramide accounted for 10–20% of the total gangliosides in human white matter. The long chain bases of individual gangliosides in human brains contained both C18 and C20 sphingosine in different ratios, and the ratio of C20 to C18 sphingosine increased with age in the order GM3<GM2<GM1<GD1a<GD1b=GT1. The ratios of individual gangliosides in the patient with Bielschowsky-Jansky disease were 2 to 3 times lower than those of age-matched controls.


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