Preservation of (3-Galactosidase Transgene in Decalcified Murine Bone Specimens Embedded in Paraffin

摘要

Genetically altered mice are an important tool for use in biomedical research. Several transgenic mice have been created whereby activation of the transgene results in the production of p-galactosidase, which can be detected with the use of histochemical methods. Although good visualization of enzyme activity in soft tissue can be complicated, it is particularly difficult in bone specimens, especially those that have been decalcified.. For example, recent studies have demonstrated the importance of canonical Wnt signaling in bone. Tcf optimal promoter (3-galactosidase (TOPGAL) or Wnt-indicator transgenic mice, in which activation of Wnt signaling results in p-galactosidase production, have been valuable tools in studying the effects of this signaling pathway in other organs,. Thus, it would be helpful to be able to detect p-galactosidase enzymatic activity in bone in these mice. In these studies, we demonstrate that, in bones taken from TOPGAL mice, p-galactosidase is better visualized when bones are decalcified and then stained with X-gal as opposed to being stained with X-gal and then being decalcified. {The J Histotechnol 31:61, 2008).