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- Title
Effect of Eimeria acervulina infection on cellspecific xanthine oxidase and inducible NO synthase activities and duodenal protein tyrosine nitration (3'-nitrotyrosine proteins) in chickens.
- Authors
Kahl, S.; Elsasser, T. H.; Miska, K. B.; Fetterer, R. H.
- Abstract
Intracellular generation of nitric oxide (NO) and superoxide anion (O2-) during proinflammatory stress can result in the formation of 3'-nitrotyrosine proteins (NTp) that correlate with alteration in protein function and metabolic impairment. Our objective was to determine the cell-specific relationship between xanthine oxidase (XO; an O2--generating enzyme), inducible NO synthase (iNOS; a NO-generating enzyme), and NTp antigens in the chicken duodenum during Eimeria acervulina infection. Ross broilers at 21 d of age were either orally infected with 3 × 105 oocysts (INF) or noninfected (NOI). At d 1, 3, 6, 7, and 10 after infection (PI), sets of 4 NOI and 4 INF birds were euthanized and blood and tissue samples collected for biochemical analysis and immunofluorescence quantification of antigens (pixel density/cell from quantitative digital image analysis; Q-dia) representative of XO, iNOS, and NTp as stratified between epithelial cells (EC; cytokeratin-18+) or infiltrating immune cells (IIC; myeloperoxidase+). Compared with NOI birds, there was a reduction in BW gain in INF birds (P < 0.01) starting at d 6. Plasma concentrations of nitrate + nitrite (estimate of systemic NO production) was increased in INF birds at d 6 and 7 (P < 0.01 vs. NOI) but normalized by d 10 PI. At the peak of infection (d 6), increases in duodenal homogenate iNOS activity were observed (P < 0.01 vs. NOI). Homogenate XO activities in INF birds were greater at d 6 PI in the liver (P < 0.05) and at d 3 and 6 PI in the ileum (P < 0.05) but not the duodenum. In contrast to homogenate iNOS and XO activities, Q-dia, as assessed by cell type, demonstrated increases (P < 0.05) in INF birds in iNOS (d 3, 6, and 7) and XO (d 1, 3, 6, and 7) proteins in EC. In IIC, XO was not detected whereas iNOS was highly expressed. Cell pixel densities associated with NTp in EC were greater in INF birds than in NOI birds (P < 0.01) at d 1, 3, 6, and 7 PI but returned to NOI levels at d 10 PI. The maximum increases in pixel densities in EC of INF birds for NTp were observed on d 6 PI (13-fold vs. NOI) and for XO were observed on d 3 PI (5-fold). Data indicate that Eimeria acervulina infection in chickens results in protein nitration in duodenal EC and that EC themselves are capable of generating the molecular signals that alone or in cooperation with IIC may ultimately lead to protein nitration.
- Subjects
EIMERIA acervulina; XANTHINE oxidase; NITROTYROSINE
- Publication
Journal of Animal Science, 2017, Vol 95, p25
- ISSN
0021-8812
- Publication type
Abstract
- DOI
10.2527/asasann.2017.051