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

D. Dolcetta, L. Perani, M. I. Givogri, F. Galbiati, A. Orlacchio, S. Martino, M. G. Roncarolo, Ernesto Bongarzone

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
Pages (from-to)462-464
Number of pages3
JournalJournal of Neuroscience Research
Volume77
Issue number3
DOIs
Publication statusPublished - Aug 1 2004

Keywords

  • Galactocerebrosidase
  • Krabbe
  • Lysosomes
  • Sphingolipids
  • Twitcher

ASJC Scopus subject areas

  • Neuroscience(all)

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