Morphometric analysis of calcium-tolerant myocytes isolated from the adult rat heart.

The ultrastructure of calcium-tolerant isolated myocytes has been compared with that of intact heart myocytes using morphometric techniques. Isolated myocytes were prepared by the method of Powell et al. [1] from four adult rat hearts, and multiple left ventricular tissue samples were obtained from a further four rat hearts after fixation by Langendorff perfusion. The subcellular-component volumes of myofibrils, mitochondria, nuclei, transverse tubules, sarcoplasmic reticulum, lipid droplets, and cytoplasmic space have been estimated using a point-counting method on thin sections of intact myocardium and isolated myocytes. No significant differences between the two sample types were found. With the use of freeze-fracture, the numerical density and distribution of sarcolemmal intramembrane particles were analyzed by the method of Jones et al. [2]. The intramembrane particle density of isolated myocyte E-faces was very similar to that of myocytes of intact myocardium. However, isolated myocytes showed a slight reduction in P-face intramembrane particle density (less than 10%), although intramembrane particle distribution remained unaltered on both fracture faces. Our findings show that the ultrastructural features of the isolated myocytes closely resemble those of their counterparts in the intact myocardium.