This chapter focuses on the rapid preparation of canine cardiac sarcolemmal vesicles by sucrose flotation. Sarcolemmal vesicles isolated from canine heart have been utilized increasingly to study the biochemical and biophysical properties of the cardiac plasma membrane. Several key activities have recently been identified in canine cardiac sarcolemmal vesicles that appear to be critically involved in regulating contractility in intact cardiac muscle. The utility of sarcolemmal vesicles for investigating biochemical mechanisms regulating ionic flux and contractility in the heart has been firmly established, and will probably continue for some time. An isolation method for preparation of sarcolemmal vesicles from canine heart, which gave substantially purified plasma membrane vesicles in reasonable yield, has been described earlier. All operations are performed at 4 °. The left ventricles and interventricular septa obtained from two dog hearts are first well-trimmed using scissors and hemostats. Endocardium, epicardium, fat, and major coronary arteries are stripped away, and the tissue is then minced into small pieces using a meat grinder.
ASJC Scopus subject areas
- Molecular Biology