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An Improved Method for Beta-Galactosidase Activity Detection on Muscle Tissue. A Light and Electron Microscopic Study

An Improved Method for Beta-Galactosidase Activity Detection on Muscle Tissue. A Light and Electron Microscopic Study

Luciana Gioglio, De Angelis M Gabriella Cusella, Renata Boratto, Paola Poggi

Ann Anat. 2002 Mar;184(2):153-7.

PMID: 11936195

Abstract:

In the present study we describe a method for the histochemical demonstration of bacterial beta-D-galactosidase activity on skeletal muscle tissue processed for light and transmission electron microscopy. Hence allowing this enzyme to be accurately detected, bacterial beta-galactosidase expression was studied in transgenic mouse where the enzyme, with the nuclear localization signal (nlacZ), is under the transcriptional control of the striated muscle-specific promoter MLC3F. The chromogenic substrate, 5-bromo-3-indolyl-beta-D-galactopyranoside (Bluo-Gal), was used both to recognize labelled myofibers, and beta-gal positive organelles inside single myofibers. Moreover, because the preservation of enzyme is highly dependent on tissue fixation, we developed a suitable fixation solution allowing good preservation of both tissue and enzymatic activity. This was achieved by briefly fixing tissue (3 hours) in glutaraldehyde (2.5%) and paraformaldehyde (1%) in combination. This method should be taken into consideration when studying the gene therapy of muscle diseases because it is sensitive, inexpensive and not time consuming.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP97753827-A	5-Bromo-3-indolyl β-D-galactopyranoside		97753-82-7	Price

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