Chromatographl C Separation of Human Brain Gangliosides
Journal of Neurochemistry 10(9): 613-623
ISSN/ISBN: 0022-3042 DOI: 10.1111/j.1471-4159.1963.tb08933.x
Gangliosides have been isolated from fetal, newborn and adult normal human brains. In the last instance gangliosides were also obtained from isolated grey and white matter. The ganglioside pattern has been studied with quantitative analytical methods and thin-layer chromatography. The resorcinol assay for sialic acids has been modified and the most common methods for the analysis of ganglioside-sialic acid have been compared. A new chromatographic method for separation of gangliosides is described. A paper roll column is used as a supporting medium and propanol-water mixtures as solvents. This type of column has been found very suitable for the separation of the gangliosides into a monosialo- and a disialoganglioside fraction and for the subsequent fractionation of the latter. By enzymic hydrolysis with sialidase it has been shown that 90 per cent of the gangliosides are derived from the same "parent ganglioside" acyl-sphingosine-N-tetrose-N-acetylneuraminic acid to which, inmost gangliosides, a further one or two moles of N-acetylneuraminic acid are bound; these are most likely bound to C3 and C6 of the galactose molecule in the terminal position. This assumption is supported by acid and enzymic hydrolysis of the di-and trisialogangliosides. In a case of infantile amaurotic idiocy (Tay-Sachs) acyl-sphingosine-N-triose-N-acetylneuraminic acid constituted more than 90 per cent of total gangliosides. Only 3-6 per cent of this ganglioside was found in normal human brains. In a case of juvenile amaurotic idiocy (Spielmeyer-Vogt) the ganglioside pattern was normal.