Analysis of apoptosis in isolated primary cardiac myocytes.
Acute myocardial infarction represents the leading cause of morbidity and mortality in the western societies. Importantly, both apoptosis and necrosis of cardiomyocytes have been implicated in the pathomechanism of myocardial infarction. The simplest way to analyze apoptosis in cardiac cells is the application of isolated neonatal primary cardiac myocytes, in which ischemia/reperfusion can be mimicked in vitro by exposing them to hypoxia and serum starvation, followed by restored oxygen and serum conditions, referred to as hypoxia/reoxygenation. In this chapter, we describe protocols routinely applied in our lab for investigating cardiomyocyte apoptosis. In summary, a better understanding of the apoptotic pathways and their regulation in the heart will potentially yield novel therapeutic targets for cardiac infarction.
Mandl
Adel
A
Boston Biomedical Research Institute, Watertown, MA, USA.
Toth
Ambrus
A
Erhardt
Peter
P
D000818:Animals; D017209:Apoptosis; D015153:Blotting, Western / Q000379:methods; D053148:Caspase 3 / Q000032:analysis / Q000378:metabolism; D002458:Cell Fractionation / Q000379:methods; D015687:Cell Hypoxia; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D045304:Cytochromes c / Q000032:analysis / Q000378:metabolism; D003584:Cytological Techniques; D007150:Immunohistochemistry / Q000379:methods; D020287:In Situ Nick-End Labeling / Q000379:methods; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology*; D010100:Oxygen / Q000378:metabolism; D051381:Rats
D045304:Cytochromes c; D053148:Caspase 3; D010100:Oxygen