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C. Cation and J. R. , This work was also supported by grants from BayGene (to P. J. O.) and the Australian Research Council (to R, J. A

E. ,

, The abbreviations used are: IDO, vol.2, p.3

T. and T. , , vol.2, p.3

D. Esi,

K. and ;. Ka,

, 3-dioxygenase-like; bNa, vol.2

, Wild type and Ido / male mice were killed by decapitation. Immunohistochemistry-After deparaffinization and rehydration, endogenous peroxidases were inhibited with 0.3% (v/v) aqueous H 2 O 2 (Sigma-Aldrich) for 30 min at ambient temperature, vol.10

M. Euromedex, PA)). After a wash in PBS, sections were incubated for 30 min with peroxidase, house rabbit polyclonal anti-IDO (1:1000) (11) was incubated overnight at 4 °C. Sections were washed for 5 min in PBS and incubated for 1 h with biotin-SP-conjugated AffiniPure goat anti-rabbit IgG (H L) antibody (1:500 in PBS/ BSA 0.1%

J. , ), and kynurenine 3-hydroxylase (EC 1.14.13.9) in mouse epididymal total RNA extracts. One g of total RNA was reverse transcribed using, ImmunoResearch Laboratories) and developed with the Vector NovaRED substrate kit for peroxidase (Vector Laboratories, vol.2, pp.kynureni- nase

, CA) 4000 QTrap mass spectrometer equipped with a turbo ion spray source. LC separation was carried out on an Atlantis T3 3-m (2.1 150-mm) reversed phase column (Waters Corp., Milford, MA) at ambient temperature using a mobile phase consisting of 0.1% formic acid in water (Solvent A) and acetonitrile (Solvent B), respectively. The gradient employed was as follows: 0-2 min, 0% B; 2-10 min, linear FIGURE 1. Scheme of mammalian tryptophan catabolism. Briefly, in mammalian cells, tryptophan is used mostly for protein synthesis. In a second quantitatively important pathway (driven by IDO in most cell types and by TDO more specifically in liver cells), it is the starting point of the kynurenine pathway. The kynurenine pathway gives birth to several metabolites, providing the appropriate enzymes that metabolize the various kynurenine intermediates are expressed. The main route of the kynurenine pathway leads to the formation of N-formyl kynurenine, L-kynurenine, 3-hydroxykynurenine, 3-hydroxyanthranilic acid, quinolinic acid, nicotinic acid, and in fine nicotinamine adenine dinucleotides. Additional lateral branches of the kynurenine pathway lead to the formation of other terminal kynurenines, such as KA, xanthurenic acid, and anthranilic acid. Kynurenines indicated in boldface type (i.e. L-kynurenine and KA) correspond to the most abundant kynurenines found in caput epididymal tissue. Outside the kynurenine pathway, tryptophan is also the precursor of serotonin and melatonin. A very small proportion of tryptophan is also transformed into indol derivatives, such as indoxyl acetic acid. Conversion of Trp to N-formyl kynurenine is achieved via IDO and/or TDO, Suppression of spermatogenesis was achieved via busulfan (1,4-butanediol dimethanesulfonate) treatment (Sigma-Aldrich). Busulfan (35 mg/kg) diluted in dimethyl sulfoxide (50%) was inoculated intraperitoneally in animals (two injections at 15 and 22 days postnatal (DPN)

, Kyn/Trp ratio in caput epididymis and serum from 6-month-old WT and Ido1 / animals.-Fold differences are indicated on the right side of the table (caput versus serum) and at the bottom of the table (Ido1 / versus WT). B and C, histograms show the levels of various inflammatory cytokines (INF-, TNF-(TNFa), interleukin-1 (IL1b), IL-3 (Il3), interleukin-6 (Il6), regulated on activation normal T cell expressed and secreted (RANTES), interleukin-12p40p70 (Il12p40p70), interleukin 4 (Il4), monocyte chemoattracting protein 1 (MCP1), and macrophage/ monocyte colony-stimulating factor (MCSF)) in caput epididymal extracts (B) and sera (C) from 6-month

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