Cysteine dioxygenase: regional localisation of protein and mRNA in rat brain.
Cysteine dioxygenase (CDO) converts cysteine to cysteinesulphinic acid and is the rate-limiting step in sulphate production. Most studies have centred upon the hepatic form of the enzyme, but several studies have investigated brain CDO using activity assays and western blotting. The aim of this study was to investigate the expression of CDO in the rat brain using a combination of immunohistochemistry and in situ hybridisation. Affinity-purified anti-R and anti-H CDO antibodies were immunoprecipitated using rat brain homogenate to determine whether the antibodies could remove enzyme activity. Immunohistochemistry and in situ hybridisation were then used to determine the cellular and regional expression of both CDO protein and mRNA. Immunoprecipitation of rat brain homogenate removed up to 98% and 70% (anti-R and anti-H, respectively) of enzyme activity. Nonimmune sheep serum had no effect upon enzyme activity. CDO protein and mRNA was localised solely to the neurones of the brain, including the pyramidal cells of the hippocampus and the Purkinje cells of the cerebellum. Regional localisation varied, with high levels of expression in the hippocampus, the dentate gyrus, the outer cortices of the brain, and the substantia nigra. The relative expression of CDO activity and protein in these regions is most probably a result of the relative abundance of neurones in these regions. CDO expression in the brain may have several possibilities functions, the most likely being the prevention of free radical production by the autoxidation of cysteine and dopamine.
Parsons
R B
RB
Department of Medicine, University of Birmingham, Queen Elizabeth Hospital, Edgbaston, Birmingham, United Kingdom. r.b.parsons.20@bham.ac.uk
Waring
R H
RH
Williams
A C
AC
Ramsden
D B
DB
D000818:Animals; D000906:Antibodies; D001921:Brain / Q000201:enzymology*; D050558:Cysteine Dioxygenase; D049308:Dioxygenases; D005609:Free Radicals / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008099:Liver / Q000201:enzymology; D008297:Male; D008969:Molecular Sequence Data; D009498:Neurotoxins / Q000378:metabolism; D010105:Oxygenases / Q000032:analysis* / Q000235:genetics* / Q000276:immunology; D011233:Precipitin Tests; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017208:Rats, Wistar
D000906:Antibodies; D005609:Free Radicals; D009498:Neurotoxins; D012333:RNA, Messenger; D010105:Oxygenases; D049308:Dioxygenases; D050558:Cysteine Dioxygenase