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Analysis of galactocerebrosidase activity in the mouse brain by a new histological staining method

✍ Scribed by D. Dolcetta; L. Perani; M.I. Givogri; F. Galbiati; A. Orlacchio; S. Martino; M.G. Roncarolo; E. Bongarzone

Publisher: John Wiley and Sons
Year: 2004
Tongue: English
Weight: 152 KB
Volume: 77
Category: Article
ISSN: 0360-4012
DOI: 10.1002/jnr.20169

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✦ Synopsis

Abstract

Gene therapy of galactocerebrosidase (GALC) deficient mice (Twitcher mutants) requires a fast and sensitive assay to detect transduced cells in vitro and in vivo. We have developed a new rapid histochemical method that specifically detects GALC activity in situ in neural cells using 5‐Br‐3Cl‐β‐galactopiranoside (X‐Gal) in the presence of taurodeoxycholic and oleic acids to enhance suspension of the substrate at low pH. Using this method, we observed robust X‐Gal staining in diverse neuronal populations and interfascicular oligodendrocytes in sections from normal mouse brain. In contrast, sections of Twitcher brain did not show a specific staining pattern in neurons or glial cells. The availability of this new sensitive and rapid in situ detection assay is fundamental for the follow‐up of Twitcher mice under gene or cellular therapies to correct central GALC deficiency. © 2004 Wiley‐Liss, Inc.

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