Identification of complement 5a-like receptor (C5L2) from astrocytes: characterization of anti-inflammatory properties.
Brain inflammation is regulated by endogenous substances, including neurotransmitters such as noradrenaline (NA), which can increase anti-inflammatory genes. To identify NA-regulated, anti-inflammatory genes, we used TOGA (total gene expression analysis) to screen rat astrocyte-derived RNA. NA-inducible cDNA clone DST11 encodes an isoform of the complement C5a receptor (C5aR), with 39% identity at the amino acid level to the rat C5aR, and 56% identity to a recently described human C5aR variant termed C5L2 (complement 5a-like receptor). Quantitative PCR confirmed that in astrocytes, DST11 mRNA expression is increased by NA, whereas in vivo depletion of cortical NA reduced DST11 levels. Western blot analysis demonstrated basal and NA-induced expression of DST11 as a 45 kDa protein in primary astrocytes cultures. Immunocytochemical staining of adult rat brain revealed DST11-immunoreactivity throughout brain, co-localized to neurons and astrocytes. In astrocytes, induction of nitric oxide synthase type 2 was increased by treatment with antisense oligonucleotides to DST11. Reducing DST11 expression also increased nuclear factor kappaB reporter gene, and decreased cAMP response element reporter gene activation. These results demonstrate that DST11 is a C5aR isoform expressed by glia and neurons, which is regulated by NA, and exerts anti-inflammatory functions. Changes in DST11 levels in diseased brain could therefore contribute to the progression of inflammatory damage.
Gavrilyuk
Vitaliy
V
Department of Anesthesiology, University of Illinois, Chicago, Illinois 60612, USA. picomol7@uic.edu
Kalinin
Sergey
S
Hilbush
Brian S
BS
Middlecamp
Andrew
A
McGuire
Susan
S
Pelligrino
Dale
D
Weinberg
Guy
G
Feinstein
Douglas L
DL
D000595:Amino Acid Sequence; D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000187:drug effects / Q000378:metabolism*; D001596:Benzylamines / Q000633:toxicity; D015153:Blotting, Western / Q000379:methods; D002478:Cells, Cultured; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism; D004347:Drug Interactions; D005786:Gene Expression Regulation / Q000187:drug effects; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D005910:Glioma; D005969:Glutamate Dehydrogenase / Q000235:genetics / Q000378:metabolism; D007150:Immunohistochemistry / Q000379:methods; D007371:Interferon-gamma / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology; D008156:Luciferases / Q000378:metabolism; D051379:Mice; D008954:Models, Biological; D016328:NF-kappa B / Q000378:metabolism; D014179:Neurotransmitter Uptake Inhibitors / Q000633:toxicity; D019001:Nitric Oxide Synthase / Q000235:genetics / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II; D009573:Nitrites / Q000378:metabolism; D009638:Norepinephrine / Q000494:pharmacology*; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D010751:Phosphopyruvate Hydratase / Q000378:metabolism; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D044087:Receptor, Anaphylatoxin C5a / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D015533:Transcriptional Activation; D014162:Transfection / Q000379:methods
D001596:Benzylamines; C496868:C5ar2 protein, rat; D017362:Cyclic AMP Response Element-Binding Protein; D005904:Glial Fibrillary Acidic Protein; D008070:Lipopolysaccharides; D016328:NF-kappa B; D014179:Neurotransmitter Uptake Inhibitors; D009573:Nitrites; D016376:Oligonucleotides, Antisense; D020033:Protein Isoforms; D012333:RNA, Messenger; D044087:Receptor, Anaphylatoxin C5a; D007371:Interferon-gamma; D008156:Luciferases; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse; C496318:Nos2 protein, rat; D005969:Glutamate Dehydrogenase; D010751:Phosphopyruvate Hydratase; C012102:DSP 4; D009638:Norepinephrine