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Journal of Histochemistry and Cytochemistry, Vol. 47, 965-968, July 1999, Copyright © 1999, The Histochemical Society, Inc.

TECHNICAL NOTE

Enhanced In Situ Detection of ß-Glucuronidase Activity in Murine Tissue

Gene C. Kopena, Darwin J. Prockopa, and Donald G. Phinneya
a Center for Gene Therapy, MCP Hahnemann University, Philadelphia, Pennsylvania

Correspondence to: Donald G. Phinney, Center for Gene Therapy, 10318 New College Bldg., 245 N. 15th St., Philadelphia, PA 19102.

We outline here a protocol for high-resolution in situ localization of ß-glucuronidase in murine tissues processed in glycol methacrylate (GMA). Murine tissues were first stained with 5-bromo-4-chloro-3-indolyl-ß-D-glucuronic acid (x-gluc), followed by histological processing in GMA. Retention of the blue indigo reaction product after overnight incubations in x-gluc allowed high-resolution localization of ß-glucuronidase activity by brightfield microscopy. When illuminated under darkfield, the x-gluc signal was enhanced, permitting detection even in cells with low-level enzyme activity. This technique offers for the first time a more sensitive enzyme histochemical method of detecting ß-glucuronidase activity in animal tissues and also the opportunity to examine expression at high magni-fication. (J Histochem Cytochem 47:965–968, 1999)

Key Words: glycol methacrylate, ß-glucuronidase, mucopolysaccharidosis type VII, x-gluc, enzyme histochemistry


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