Distribution of cholesterol and galactosylceramide in rat cerebellar white matter.
White matter and the inner granular layer of rat cerebellum was analysed by imaging time-of-flight secondary-ion mass spectrometry (TOF-SIMS) equipped with a Bi+ ion cluster gun. Samples were prepared by high pressure freezing, freeze-fracturing and freeze drying or by plunge freezing and cryostat sectioning. The identified and localized chemical species were: sodium, potassium, phosphocholine, cholesterol and galactosylceramide (GalC) with carbon chain lengths C18:0 (N-stearoyl-galactosylceramide) and C24:0 (N-lignoceroylgalactosylceramide) with CH24:0 (hydroxy-lignoceroylgalactosylceramide). We report new findings regarding the organization of myelin in white matter. One is cholesterol-rich, ribbon-shaped 10-20 microm areas excluding Na+ and K+. The second finding is the different distribution of GalC C18 and GalC C24 in relation to these areas, where GalC C18 was localized in cholesterol-rich areas and GalC C24 was localized in Na/K-enriched areas. The distribution of GalC was in small spots, homogeneous in size, of 0.8-1.5 microm. Sample preparation with high pressure freezing allowed separate localization of sodium and potassium in tissue samples.
Börner
Katrin
K
Department of Anatomy and Cell Biology, University of Göteborg, Box 420, SE 405 30 Göteborg, Sweden.
Nygren
Håkan
H
Hagenhoff
Birgit
B
Malmberg
Per
P
Tallarek
Elke
E
Månsson
Jan-Eric
JE
D000818:Animals; D002531:Cerebellum / Q000033:anatomy & histology / Q000737:chemistry / Q000378:metabolism; D002784:Cholesterol / Q000737:chemistry / Q000378:metabolism*; D005699:Galactosylceramides / Q000737:chemistry / Q000378:metabolism*; D008297:Male; D013058:Mass Spectrometry; D009186:Myelin Sheath / Q000737:chemistry*; D051381:Rats; D017207:Rats, Sprague-Dawley
D005699:Galactosylceramides; D002784:Cholesterol