| Identification of a novel GDNF mRNA induced by LPS in immune cell lines. | |
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MedLine Citation:
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PMID: 18295922 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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. |
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Authors:
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Tatsuhide Tanaka; Kentaro Oh-hashi; Masatoshi Ito; Hironobu Shitara; Yoko Hirata; Kazutoshi Kiuchi |
Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2008-01-17 |
Journal Detail:
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Title: Neuroscience research Volume: 61 ISSN: 0168-0102 ISO Abbreviation: Neurosci. Res. Publication Date: 2008 May |
Date Detail:
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Created Date: 2008-04-21 Completed Date: 2008-07-22 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 8500749 Medline TA: Neurosci Res Country: Ireland |
Other Details:
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Languages: eng Pagination: 11-7 Citation Subset: IM |
Affiliation:
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Department of Biomolecular Science, Faculty of Engineering, Gifu University, 1-1 Yanagido, Gifu, Japan. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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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:
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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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