Journal
JOURNAL OF NEUROIMMUNOLOGY
Volume 174, Issue 1-2, Pages 52-62Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jneuroim.2006.01.006
Keywords
dynorphin; prodynorphin; U-937 cells; lipopolysaccharide; gene expression
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Funding
- NIDA NIH HHS [K05-DA00360] Funding Source: Medline
- DS NIH HHS [DS04355] Funding Source: Medline
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Dynorphins are endogenous kappa opioid peptides widely distributed in the central nervous system. A number of recent reports have established their roles in modulating immunological functions. In this study, we investigated the expression of prodynorphin (PDYN) gene, which encodes the precursor of dynorphin peptides, in human macrophage U-937 cell line. PDYN mRNA was expressed at a detectable level, measured with a standard RT-PCR method in U-937 cells, but not in Jurkat and Raji cells, which are human T and B lymphocytes, respectively. Further analyses with RT-PCR assays by using primers covering each exon of the PDYN gene showed that U-937 cells expressed the adult brain-type PDYN mRNA. Most interestingly, activation of U-937 cells with lipopolysaccharide (LPS) led to a decrease in PDYN mRNA levels. This decrease was dependent on both the concentration of LPS and the duration of LPS treatment. In order to test the role of transcription in LPS-mediated down-regulation of PDYN gene expression in U937 cells, the proximal PDYN gene promoter with a length of either similar to 300 base pairs or similar to 900 base pairs was cloned and inserted upstream of luciferase reporter gene. Results from transient transfections and dual luciferase assays showed that PDYN gene promoter activity was decreased by LPS treatment. Taken together, our results suggested the U-937 cells expressed adult brain-type PDYN mRNA which was down-regulated by activation of the cells with LPS due to an inhibition of PDYN gene transcription. (c) 2006 Elsevier B.V All rights reserved.
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