A quantitative study of peroxidase activity in unfixed tissue sections of the guinea-pig thyroid gland.
A technique for the cytochemical demonstration of peroxidase activity in unfixed guinea-pig thyroid tissue is described in this paper. The substrate 3,3'-diaminobenzidine tetrahydrochloride (DAB) is oxidized by the peroxidase to form an insoluble reaction product. Optimal results were obtained after 20 min incubation at 37 degrees C in reaction medium containing 1.4 mM DAB (in 0.1 M Tris-HCl) and 0.15 mM hydrogen peroxide at pH 8.0. Peroxidase activity was seen in the thyroid follicle cells as a diffuse brown reaction product (which was more dense and granular in erythrocytes). The enzyme activity was quantified using a scanning-integrating microdensitometer, and the effects of two specific peroxidase inhibitors were evaluated. Both 3-amino-1,2,4-triazole and methimazole inhibited peroxidase activity in the follicle cells (enzyme activity was still seen in the erythrocytes), maximal inhibition occurring at 10 mM. Stimulation of peroxidase in the thyroid was observed in vivo (1 I.U. TSH administered every 8 h for two days), with the maximal stimulation occurring after 1 day.
D015100:3,3'-Diaminobenzidine / Q000378:metabolism; D000640:Amitrole / Q000494:pharmacology; D000818:Animals; D005260:Female; D006168:Guinea Pigs; D006861:Hydrogen Peroxide / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007453:Iodide Peroxidase / Q000032:analysis* / Q000037:antagonists & inhibitors / Q000378:metabolism; D008713:Methimazole / Q000494:pharmacology; D010544:Peroxidases / Q000032:analysis*; D013961:Thyroid Gland / Q000187:drug effects / Q000201:enzymology*; D013972:Thyrotropin / Q000494:pharmacology
D015100:3,3'-Diaminobenzidine; D008713:Methimazole; D013972:Thyrotropin; D006861:Hydrogen Peroxide; D010544:Peroxidases; D007453:Iodide Peroxidase; D000640:Amitrole