Analysis of fiber-type differences in reporter gene expression of β-gal transgenic muscle

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 798; p. 445
• Main Authors: Tai, Phillip W L, Smith, Catherine L, Angello, John C, Hauschka, Stephen D
• Format: Journal Article
• Language: English
• Published: United States 2012
• Subjects: Animals; Antibodies, Monoclonal - biosynthesis; beta-Galactosidase - analysis; beta-Galactosidase - genetics; Frozen Sections - methods; Genes, Reporter; Mice; Mice, Transgenic; Muscle Fibers, Skeletal - cytology; Muscle Fibers, Skeletal - metabolism; Staining and Labeling - methods
• ISSN: 1940-6029
• DOI: 10.1007/978-1-61779-343-1_26

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.