Document Detail

Induction of axonal differentiation by silencing plasma membrane-associated sialidase Neu3 in neuroblastoma cells.
MedLine Citation:
PMID:  17176265     Owner:  NLM     Status:  MEDLINE    
A reduction of 70% of the plasma membrane-associated sialidase Neu3 activity, due to a corresponding reduction of the enzyme expression by transducing cells with a short hairpin RNA encoding a sequence target (complementary messenger of mouse Neu3), caused neurite elongation in Neuro2a murine neuroblastoma cells. The differentiation process was accompanied in parallel by an increase of the acetylcholinesterase activity, a moderate increase of the c-Src expression and by the presence of the axonal marker tau protein on the neurites. The sphingolipid pattern and turnover in transduced and control cells were characterized by thin layer chromatography, mass spectrometry and metabolic radiolabeling after feeding cells with tritiated sphingosine. Control cells contained about 2 nmol of gangliosides/mg cell protein. GM2 was the main compound, followed by GD1a, GM3 and GM1. In Neu3 silenced cells, the total ganglioside content remained quite similar, but GM2 increased by 54%, GM3 remain constant, and GM1 and GD1a decreased by 66% and 50%, respectively. Within the organic phase sphingolipids, ceramide decreased by 50%, whereas the sphingomyelin content did not change in Neu3 silenced cells.
Rea Valaperta; Manuela Valsecchi; Federica Rocchetta; Massimo Aureli; Simona Prioni; Alessandro Prinetti; Vanna Chigorno; Sandro Sonnino
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2006-12-14
Journal Detail:
Title:  Journal of neurochemistry     Volume:  100     ISSN:  0022-3042     ISO Abbreviation:  J. Neurochem.     Publication Date:  2007 Feb 
Date Detail:
Created Date:  2007-01-31     Completed Date:  2007-03-20     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  2985190R     Medline TA:  J Neurochem     Country:  England    
Other Details:
Languages:  eng     Pagination:  708-19     Citation Subset:  IM    
Department of Medical Chemistry, Biochemistry and Biotechnology, and Center of Excellence on Neurodegenerative Diseases, University of Milan, Segrate, Italy.
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MeSH Terms
Acetylcholinesterase / metabolism
Cell Differentiation / genetics*
Cell Line, Tumor
Ceramides / metabolism
Down-Regulation / genetics
Gangliosides / metabolism
Gene Silencing / physiology
Growth Cones / metabolism*,  ultrastructure
Membrane Proteins / genetics*
Nervous System / cytology,  growth & development*,  metabolism*
Neuraminidase / genetics*
RNA / genetics
Sphingolipids / metabolism
Transduction, Genetic
src-Family Kinases / metabolism
tau Proteins / metabolism
Reg. No./Substance:
0/Ceramides; 0/Gangliosides; 0/Membrane Proteins; 0/Sphingolipids; 0/tau Proteins; 63231-63-0/RNA; EC Kinases; EC; EC protein, mouse; EC

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