title: Analysis of fiber-type differences in reporter gene expression of β-gal transgenic muscle.
abstract: β-galactosidase (β-gal) is among the most frequently used markers for studying a wide variety of biological mechanisms, e.g., gene expression, cell migration, stem cell conversion to different cell types, and gene silencing. Many of these studies require the histochemical detection of relative β-gal levels in tissue cross-sections mounted onto glass slides and visualized by microscopy. This is particularly useful for the analysis of promoter activity in skeletal muscle tissue since the β-gal levels can vary dramatically between different anatomical muscles and myofiber types. The differences in promoter activity can be due to a myofiber's developmental history, innervation, response to normal or experimental physiological signals, and its disease state. It is thus important to identify the individual fiber types within muscle cross-sections and to correlate these with transgene expression signals. Here, we provide a detailed description of how to process and analyze muscle tissues to determine the fiber-type composition and β-gal transgene expression within cryosections.
authors: Department of Biochemistry, University of Washington, Seattle, WA, USA.
authors: Phillip W L
authors: PW
authors: Tai
authors: Catherine L
authors: CL
authors: Smith
authors: John C
authors: JC
authors: Angello
authors: Stephen D
authors: SD
authors: Hauschka
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D005629:Frozen Sections / Q000379:methods; D017930:Genes, Reporter; D051379:Mice; D008822:Mice, Transgenic; D018485:Muscle Fibers, Skeletal / Q000166:cytology / Q000378:metabolism*; D013194:Staining and Labeling / Q000379:methods; D001616:beta-Galactosidase / Q000032:analysis* / Q000235:genetics
chemical_list: D000911:Antibodies, Monoclonal; D001616:beta-Galactosidase