title: Analysis of galactocerebrosidase activity in the mouse brain by a new histological staining method.
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-beta-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.
authors.affiliation: San Raffaele Telethon Institute for Gene Therapy, Milano, Italy.
authors.forename: D
authors.initials: D
authors.lastname: Dolcetta
authors.forename: L
authors.initials: L
authors.lastname: Perani
authors.forename: M I
authors.initials: MI
authors.lastname: Givogri
authors.forename: F
authors.initials: F
authors.lastname: Galbiati
authors.forename: A
authors.initials: A
authors.lastname: Orlacchio
authors.forename: S
authors.initials: S
authors.lastname: Martino
authors.forename: M G
authors.initials: MG
authors.lastname: Roncarolo
authors.forename: E
authors.initials: E
authors.lastname: Bongarzone
mesh_terms: D000818:Animals; D001921:Brain / Q000201:enzymology*; D004789:Enzyme Activation; D005698:Galactosylceramidase / Q000032:analysis* / Q000172:deficiency / Q000378:metabolism*; D006651:Histocytochemistry / Q000379:methods; D051379:Mice; D008810:Mice, Inbred C57BL; D008818:Mice, Neurologic Mutants; D009457:Neuroglia / Q000201:enzymology; D009474:Neurons / Q000201:enzymology; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D005698:Galactosylceramidase