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Abstract
This report describes a new method for the microanalysis of sphingolipids and its
application for the characterization of cerebrosides and sulfatides in multiple sclerosis
brain and rat sciatic nerves undergoing Wallerian degeneration. Tissue was extracted
with isopropanol/hexane (20 : 78), and the total lipids obtained were subjected to
benzoylation-desulfation. A portion of this was directly analyzed by silica-column
high performance liquid chromatography for the determination of nonhydroxycerebroside,
hydroxycerebroside, nonhydroxysulfatide, and hydroxysulfatide. Another portion was
fractionated by thin-layer chromatography, and the spots corresponding to the sphingolipid
derivatives were eluted. The material from each spot was analyzed by reverse phase
high performance liquid chromatography for its homolog composition. With this new
procedure the concentrations and homolog compositions of cerebrosides and sulfatides
were measured in plaque, periplaque, and normal-appearing white matter from brains
of multiple sclerosis patients and Wallerian degenerated rat sciatic nerves distal
to the nerve transection. One piece of plaque studied contained only 1.86, 2.76, 0.60,
and 0.45 nmol of nonhydroxycerebroside, hydroxycerebroside, nonhydroxysulfatide and
hydroxysulfatide/mg of protein, respectively. These concentrations are less than 1%
of those found in normal white matter. Periplaques were found to contain concentrations
of these sphingolipids between those of plaque and normal white matter. The levels
of these sphingolipids in degenerative nerves were 10–20% below normal the third day
after the nerve was severed and about 70% below normal after 10 days. The rate of
decrease lessened from ten days to 55 days. The homolog compositions of these sphingolipids
in both multiple sclerosis brain and degenerating nerves were similar to those in
the control. The implications of these findings and the advantages of this new analytical
method are discussed.
Abbreviations:
HPLC (high performance liquid chromatography), TLC (thin-layer chromatography)To read this article in full you will need to make a payment
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Article info
Publication history
Accepted:
November 18,
1981
Received in revised form:
November 16,
1981
Received:
September 28,
1981
Footnotes
☆This investigation was supported in part by Research Grants NS-13559, NS-13569, and HD-10981 from the National Institutes of Health, BNS 8022444 from the National Science Foundation, and grants from the National Multiple Sclerosis Society and the Central Office Research Funds of the Veterans Administration Hospital.
Identification
Copyright
© 1982 Published by Elsevier Inc.