Simultaneous in situ hybridization and TUNEL to identify cells undergoing apoptosis |
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Authors: | Bernadette R Gochuico Mary C Williams Alan Fine |
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Institution: | (1) The Pulmonary Center and Department of Biochemistry and Anatomy, Boston University School of Medicine, 80 E. Concord Street, R-3, Boston, MA 02118, USA |
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Abstract: | Apoptotic cells in tissue sections can be localized by in situ labelling of partly degraded DNA. In a heterogeneous population of cells, however, the specific identity of cell types undergoing
apoptosis often cannot be reliably achieved at the light microscope level because of the marked alterations in cellular morphology
that characterize apoptosis. In order to clearly specify cell types undergoing apoptosis, in situ end labelling has been coupled to immunohistochemistry. This method is limited by the availability of antibodies that bind
to cell-specific protein markers in tissue sections. In contrast, we describe a method that combines in situ end labelling
with in situ hybridization, a technique that specifies cell types based on mRNA expression. Taking advantage of the specific expression
of surfactant protein C mRNA in type II alveolar epithelial cells, we demonstrate that this technique has the ability to localize
alveolar type II cells undergoing apoptosis in vivo after the intratracheal instillation of an antibody that activates the cell surface Fas protein. The wide availability of
cell-specific gene markers suggests that this method can be adapted to define cell types that undergo apoptosis during various
physiological and pathological states in vivo.
This revised version was published online in November 2006 with corrections to the Cover Date. |
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