In situ detection of spontaneous superoxide anion and singlet oxygen production by mitochondria in rat liver and small intestine Kerver, E.D.; Vogels, I.M.C.; Bosch, K.S.; Vreeling-Sindelárová, H.; van den Munckhof, R.J.M.; Frederiks, W.M. Published in:The histochemical journal DOI:10.1023/A:1026453926517 Link to publication Citation for published version (APA):Kerver, E. D., Vogels, I. M. C., Bosch, K. S., Vreeling-Sindelárová, H., van den Munckhof, R. J. M., & Frederiks, W. M. (1997). In situ detection of spontaneous superoxide anion and singlet oxygen production by mitochondria in rat liver and small intestine. The histochemical journal, 29, 229-237. DOI: 10.1023/A:1026453926517 General rightsIt is not permitted to download or to forward/distribute the text or part of it without the consent of the author(s) and/or copyright holder(s), other than for strictly personal, individual use, unless the work is under an open content license (like Creative Commons). Disclaimer/Complaints regulationsIf you believe that digital publication of certain material infringes any of your rights or (privacy) interests, please let the Library know, stating your reasons. In case of a legitimate complaint, the Library will make the material inaccessible and/or remove it from the website. Please Ask the Library: http://uba.uva.nl/en/contact, or a letter to: Library of the University of Amsterdam, Secretariat, Singel 425, 1012 WP Amsterdam, The Netherlands. You will be contacted as soon as possible. Download date: 11 May 2018In situ detection of spontaneous superoxide anion and singlet oxygen production by mitochondria in rat liver and small intestine SummaryIn the present study, the endogenous formation of reactive oxygen species was localized in rat liver and small intestine. The 3,39-diaminobenzidine (DAB)-Mn 2 technique in which cobalt ions were included in the incubation medium was applied to unfixed cryostat sections of intact tissues. Addition of manganese ions to the DAB-Co 2 -containing medium greatly increased the amounts of final reaction product formed compared with incubations with only DAB and cobalt ions. In liver, a blue final reaction product was deposited, particularly in hepatocytes surrounding portal tracts. In the small intestine, the DAB-cobalt complex was mainly found at the basal side of enterocytes. Goblet cells remained unstained. Electron microscopical images revealed that an electron-dense reaction product was exclusively present at both inner and outer membranes and at the intermembrane space in mitochondria of liver parenchymal cells and duodenal enterocytes. It was shown that the spontaneous formation of final reaction product was enzymatic and dependent on the presence of oxygen in the medium. Sulphide decreased the reaction, which may indicate that cytochrome c oxidase was partially involved. Benzoquinone and histidine, which are scavengers of superoxide anions and singlet oxygen respectively, reduced the amount of final reaction product considerably. Furthermore, the formation of final reaction pro...