Document Detail

Localization of cholesterol, phosphocholine and galactosylceramide in rat cerebellar cortex with imaging TOF-SIMS equipped with a bismuth cluster ion source.
MedLine Citation:
PMID:  16300993     Owner:  NLM     Status:  MEDLINE    
Time-of-flight secondary-ion-mass-spectrometry (TOF-SIMS) was utilized to address the issue of co-localization of cholesterol, phosphocholine and galactosylceramide in rat cerebellar cortex. Rat cerebellum was fixed, freeze-protected by sucrose, frozen and sectioned by cryoultramicrotomy and dried at room temperature. The samples were analyzed in an imaging TOF-SIMS instrument equipped with a Bi(1-7)+-source. The cholesterol signal (m/z 369 and 385) was localized in Purkinje cells and in nuclei of granular layer cells. The phosphocholine headgroup of phosphatidylcholine and sphingomyelin was localized by imaging a specific fragment (m/z 86). This signal was localized in the molecular layer of cerebellar cortex, in Purkinje cells and in parts of the granular layer probably representing the synapse-rich glomeruli. The galactosylceramide was localized by imaging the quasi-molecular ions at m/z 835 and 851, showed a clear colocalization with cholesterol, but also a specific localization in dots (diameter <or=700 nm) in the molecular layer in the vicinity of Purkinje cells, at Purkinje cells and at cells in the granular layer. The results show a heterogeneous distribution of lipids between different cell types not previously described. In order to avoid redistribution artefacts, controls were made by a technique, based on high pressure freezing ,freeze fracturing and freeze drying of samples which were then analyzed by bombardment with a Bi3+ liquid metal ion gun. The galactosylceramide and cholesterol were found distributed as spots in the granular layer. The spots were of homogeneous size with a diameter of <700 nm. Although the galactosylceramide and cholesterol were localized to the same areas, there were clear differences in their distribution at higher resolution.
H?kan Nygren; Katrin B?rner; Birgit Hagenhoff; Per Malmberg; Jan-Eric M?nsson
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2005-11-08
Journal Detail:
Title:  Biochimica et biophysica acta     Volume:  1737     ISSN:  0006-3002     ISO Abbreviation:  Biochim. Biophys. Acta     Publication Date:  2005 Dec 
Date Detail:
Created Date:  2005-12-12     Completed Date:  2006-02-13     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0217513     Medline TA:  Biochim Biophys Acta     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  102-10     Citation Subset:  IM    
Department of Anatomy and Cell Biology, University of G?teborg, P.O. Box 420, SE-405 30 G?teborg, Sweden.
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MeSH Terms
Cerebellar Cortex / metabolism*
Cholesterol / metabolism*
Galactosylceramides / metabolism*
Phosphorylcholine / metabolism*
Purkinje Cells / metabolism
Rats, Sprague-Dawley
Spectrometry, Mass, Secondary Ion / methods*
Tissue Distribution
Reg. No./Substance:
0/Galactosylceramides; 107-73-3/Phosphorylcholine; 57-88-5/Cholesterol; 7440-69-9/Bismuth

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