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Nitric oxide synthesis by the chicken macrophage (HD‐11) cell line results in altered gene expression of multiple arginine transport systems
Author(s) -
Moulds Michael B,
Humphrey Brooke D
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.907.13
Subject(s) - lipopolysaccharide , nitric oxide , nitrite , nitric oxide synthase , messenger rna , chemistry , microbiology and biotechnology , arginine , cell culture , macrophage , transporter , biochemistry , biology , gene , endocrinology , amino acid , nitrate , in vitro , genetics , organic chemistry
Macrophages metabolize arginine (ARG) to nitric oxide (NO) via inducible nitric oxide synthase (iNOS). In mammals, macrophage NO synthesis is dependent upon CAT‐2B mediated ARG uptake; however, in avian macrophages the transporter(s) coordinating ARG uptake for NO synthesis are unknown. Therefore, HD‐11 cells were cultured in complete media with 0 (control) or 1μg/ml E. coli lipopolysaccharide (LPS) to determine any changes in the ARG transporter(s) involved in ARG uptake for NO production. At 24 and 48 h of culture, media nitrite concentrations were measured and ARG transporter systems and iNOS mRNA abundance were measured by quantitative real‐time PCR. LPS increased nitrite 18‐fold and 76‐fold at 24 and 48 h, respectively, compared to control (P<0.05). LPS increased iNOS mRNA 8.5‐fold compared to control at 48 h (P<0.05). CAT‐2B mRNA was not detected in control or LPS treated HD‐11 cells. LPS increased the mRNA abundance of ARG importers CAT‐2A by 1.6‐fold and b0,+ by 3.7‐fold at 48 h. LPS induced mRNA expression of ARG importers CAT‐1 and CAT‐3 at 48 h. LPS decreased the mRNA abundance of the ARG exporter y+LAT1 by 71% at 48 h. Unlike mammals, avian macrophages appear to utilize a number of ARG transport systems to increase ARG uptake for NO production. Funding: California Agricultural Research Initiative