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DOI: 10.1369/jhc.4A6505.2005
Volume 53 (5): 557-570, 2005
Copyright ©The Histochemical Society, Inc.
Tissue-specific Tropomyosin Isoform Composition
Oncology Research Unit (GS,BV,NSB,SE,GMO,PWG) and Discipline of Paediatrics and Child Health (GS,GMO,PWG), University of Sydney, Sydney, Australia; John Douglas Centre for Structural Pathology (MRQ) and Institute for Neuromuscular Research (NY), The Children's Hospital at Westmead, Sydney, Australia; and Children's Cancer Institute Australia for Medical Research, Randwick, Australia (NMV,MK)
Correspondence to: Peter Gunning, Oncology Research Unit, The Children's Hospital at Westmead, Locked Bag 4001, Westmead, NSW, Sydney, Australia. E-mail: peterg3{at}chw.edu.au
Four distinct genes encode tropomyosin (Tm) proteins, integral components of the actin microfilament system. In non-muscle cells, over 40 Tm isoforms are derived using alternative splicing. Distinct populations of actin filaments characterized by the composition of these Tm isoforms are found differentially sorted within cells (Gunning et al. 1998b). We hypothesized that these distinct intracellular compartments defined by the association of Tm isoforms may allow for independent regulation of microfilament function. Consequently, to understand the molecular mechanisms that give rise to these different microfilaments and their regulation, a cohort of fully characterized isoform-specific Tm antibodies was required. The characterization protocol initially involved testing the specificity of the antibodies on bacterially produced Tm proteins. We then confirmed that these Tm antibodies can be used to probe the expression and subcellular localization of different Tm isoforms by Western blot analysis, immunofluorescence staining of cells in culture, and immunohistochemistry of paraffin wax–embedded mouse tissues. These Tm antibodies, therefore, have the capacity to monitor specific actin filament populations in a range of experimental systems. (J Histochem Cytochem 53:557–570, 2005)
Key Words: non-muscle tropomyosin • actin • immunohistochemistry • cytoskeleton
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