Document Detail

Altered interaction and expression of proteins involved in neurosecretion in scrapie-infected GT1-1 cells.
MedLine Citation:
PMID:  15528199     Owner:  NLM     Status:  MEDLINE    
Prions cause transmissible and fatal diseases that are associated with spongiform degeneration, astrogliosis, and loss of axon terminals in the brains. To determine the expression of proteins involved in neurosecretion and synaptic functions after prion infection, gonadotropin-releasing hormone neuronal cell line subclone (GT1-1) was infected with the RML scrapie strain and analyzed by Western blotting, real time PCR, and immunohistochemistry. As revealed by Western blotting of lysates exposed to different temperatures, the levels of complexed SNAP-25, syntaxin 1A, and synaptophysin were decreased in scrapie-infected GT1-1 cells (ScGT1-1), whereas the level of monomeric forms of these proteins was increased and correlated to the level of scrapie prion protein (PrPSc). However, when complex formation was prevented by prolonged heating of samples in SDS, the levels of monomeric SNAP-25, syntaxin 1A and synaptophysin in ScGT1-1 cells were decreased in comparison to GT1-1 cells. The reduced level of SNAP-25 was observed as early as 32 days postinfection. Increased mRNA levels of both splice variants SNAP-25a and -b in ScGT1-1 cells were seen. No difference in the morphology, neuritic outgrowth or distribution of SNAP-25, syntaxin 1A, or synaptophysin could be observed in ScGT1-1 cells. Treatment with quinacrine or pentosan polysulfate cleared the PrPSc from the ScGT1-1 cell cultures, and the increase in levels of monomeric SNAP-25 and synaptophysin was reversible. These results indicate that a scrapie infection can cause changes in the expression of proteins involved in neuronal secretion, which may be of pathogenetic relevance for the axon terminal changes seen in prion-infected brains.
Malin K Sandberg; Peter Löw
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.     Date:  2004-11-04
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  280     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  2005 Jan 
Date Detail:
Created Date:  2005-01-11     Completed Date:  2005-02-25     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1264-71     Citation Subset:  IM    
Department of Neuroscience, Karolinska Institutet, Retzius väg 8 B2: 5, Stockholm, S-171 77, Sweden.
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MeSH Terms
1-Methyl-3-isobutylxanthine / pharmacology
Antigens, Surface / metabolism
Cell Line
Cyclic AMP / analogs & derivatives,  pharmacology
Membrane Proteins / genetics,  metabolism
Nerve Tissue Proteins / genetics,  metabolism
Neurons / drug effects,  metabolism*,  pathology*,  secretion
Pentosan Sulfuric Polyester / pharmacology
PrPSc Proteins / metabolism
Protein Binding
Protein Isoforms / genetics,  metabolism
Quinacrine / pharmacology
RNA, Messenger / genetics,  metabolism
Scrapie / metabolism*,  pathology*
Synaptophysin / metabolism
Synaptosomal-Associated Protein 25
Syntaxin 1
Time Factors
Reg. No./Substance:
0/Antigens, Surface; 0/Membrane Proteins; 0/Nerve Tissue Proteins; 0/PrPSc Proteins; 0/Protein Isoforms; 0/RNA, Messenger; 0/Snap25 protein, mouse; 0/Stx1a protein, mouse; 0/Synaptophysin; 0/Synaptosomal-Associated Protein 25; 0/Syntaxin 1; 28822-58-4/1-Methyl-3-isobutylxanthine; 37300-21-3/Pentosan Sulfuric Polyester; 60-92-4/Cyclic AMP; 83-89-6/Quinacrine

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