Improved in situ beta-galactosidase staining for histological analysis of transgenic mice.
The present study describes a novel method for the histochemical demonstration of beta-galactosidase activity on tissue sections. We have replaced 5-bromo-4-chloro-3-indolyl-beta-D-galactoside (X-Gal) with 5-bromo-indolyl-beta-o-galactopyranoside (Bluo-Gal) as a chromogenic substrate for the bacterial beta-galactosidase (lacZ). After beta-galactosidic cleavage, Bluo-Gal precipitates in form of fine birefringent crystals, whereas X-gal gives rise to an amorphous precipitate. Upon microscopic examination under polarized light, the crystals emit a strong signal consisting of yellow reflected light. This property of Bluo-Gal results in greatly enhanced sensitivity of the staining method for beta-galactosidase and allows for optimal morphological resolution. To exemplify the applications of this technique, the expression is demonstrated in transgenic mice of beta-galactosidase driven by a fragment of the human tissue-type plasminogen activator promoter.
Aguzzi
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A
Institute of Neuropathology, Zurich, Switzerland.
Theuring
F
F
D000818:Animals; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D006651:Histocytochemistry; D017403:In Situ Hybridization; D007763:Lac Operon; D051379:Mice; D008822:Mice, Transgenic; D008867:Microtomy; D013194:Staining and Labeling / Q000379:methods*; D010959:Tissue Plasminogen Activator / Q000235:genetics; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism*
D001616:beta-Galactosidase; D010959:Tissue Plasminogen Activator