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Journal of Histochemistry and Cytochemistry, Vol. 51, 865-871, July 2003, Copyright © 2003, The Histochemical Society, Inc.

ARTICLE

Quantitative Histochemical Assay for Superoxide Dismutase in Rat Brain

Alessandro Viggianoa, Davide Viggianoa, Andrea Viggianoa, and Bruno De Lucaa
a Department of Experimental Medicine, Second University, Naples, Italy

Correspondence to: Bruno De Luca, Dept. of Experimental Medicine, Seconda Università degli Studi di Napoli Via Costantinopoli, 16, 80138 Naples, Italy. E-mail: bruno.deluca@unina2.it

Superoxide anions are highly reactive radicals overproduced in many pathological situations such as inflammation and ischemia. One of the major factors in the protection from superoxide anions is the enzyme superoxide dismutase (SOD), which catalyzes the dismutation of superoxide to hydrogen peroxide. This study presents a quantitative histochemical method to estimate SOD activity in rat brain tissue sections. This method is based on the cerium capture method and 3,3'-diaminobenzidine amplification of transition cerium compounds. Substrate for SOD was provided by reduction of oxygen during the autoxidation of riboflavin in the presence of UV light. This histochemical method reveals the overall activity of the three different forms of SOD described in mammalian tissues: cytosolic copper–zinc SOD, mitochondrial manganese SOD, and the high molecular weight extracellular SOD. Eventually, this method can be used to quantify SOD activity in tissue sections by image analysis. (J Histochem Cytochem 51:865–871, 2003)

Key Words: free radicals, cerium, DAB, quantitative histochemistry, image analysis


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