Document Detail


Identification of a novel GDNF mRNA induced by LPS in immune cell lines.
MedLine Citation:
PMID:  18295922     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Glial cell line-derived neurotrophic factor (GDNF) is a survival factor for many neuronal cell types and has wide ranging effects within the central nervous system. To investigate the expression of the GDNF gene in immune cell lines under inflammatory conditions, we pharmacologically estimated the induction of GDNF mRNA in RAW264.7 cells. RT-PCR analysis revealed that LPS-induced GDNF mRNA in RAW264.7 cells does not include exon 3, which encodes the translational start site of this gene. A novel type of GDNF mRNA cloned by 5'-RACE consisted of the previous exon 4 and its flanking 5' upstream region, akin to a single exon gene. A similar type of human GDNF mRNA was also detected in a human neuroblastoma cell line, SH-SY5Y, without any stimulation. This novel (Ex4) GDNF mRNA was also upregulated by LPS in primary cultured rat macrophages, microglia and astrocytes and was found to exist in mouse brain. Ex4 GDNF protein produced by transfected HEK293 cells was mainly detected in cell lysates, but in conditioned medium only after PMA stimulation. Ex4 GDNF protein was found to exist as an unglycosylated form in both the transfected cells and the conditioned medium while full-type GDNF protein is glycosylated. PMA-stimulated augmentation of unglycosylated Ex4 GDNF protein was demonstrably regulated at the post-translational level.
Authors:
Tatsuhide Tanaka; Kentaro Oh-hashi; Masatoshi Ito; Hironobu Shitara; Yoko Hirata; Kazutoshi Kiuchi
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2008-01-17
Journal Detail:
Title:  Neuroscience research     Volume:  61     ISSN:  0168-0102     ISO Abbreviation:  Neurosci. Res.     Publication Date:  2008 May 
Date Detail:
Created Date:  2008-04-21     Completed Date:  2008-07-22     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8500749     Medline TA:  Neurosci Res     Country:  Ireland    
Other Details:
Languages:  eng     Pagination:  11-7     Citation Subset:  IM    
Affiliation:
Department of Biomolecular Science, Faculty of Engineering, Gifu University, 1-1 Yanagido, Gifu, Japan.
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MeSH Terms
Descriptor/Qualifier:
Animals
Base Sequence
Blotting, Western
Cell Line
Cloning, Molecular
DNA, Complementary / biosynthesis,  genetics
Exons / genetics
Glial Cell Line-Derived Neurotrophic Factor / biosynthesis*,  genetics
Glucuronidase / pharmacology
Humans
Lipopolysaccharides / pharmacology*
Macrophages / drug effects,  metabolism*
Mice
Molecular Sequence Data
Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / pharmacology
RNA, Messenger / biosynthesis*,  genetics
Rats
Rats, Sprague-Dawley
Reverse Transcriptase Polymerase Chain Reaction
Transfection
Chemical
Reg. No./Substance:
0/DNA, Complementary; 0/Glial Cell Line-Derived Neurotrophic Factor; 0/Lipopolysaccharides; 0/RNA, Messenger; EC 3.2.1.-/heparanase; EC 3.2.1.31/Glucuronidase; EC 3.5.1.52/Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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