Postnatal growth of cardiomyocytes in the left ventricle of the rat.

Abstract

We studied the development of myocytes and interstitium using perfusion-fixed left ventricles obtained from normal female Wistar rats at 5 days (n = 5), 25 days (n = 5), and 125 days (n = 5) of age. Using design-based stereological methods and light microscopy, we estimated the following parameters: volume of left ventricle made up by myocytes, myocyte nuclei, and interstitium; total numbers of myocyte and non-myocyte nuclei; mean volumes of myocyte nuclei; the total volume, surface area, and length of fibers; and the mean star volumes of fibers. Some derived parameters were also calculated, namely, the mean myocardium volume per nucleus and the mean fiber cross-sectional area. We found that postnatal myocyte growth after day 5 in the young rat is largely hypertrophic, while interstitial growth is hyperplastic. The increase in left ventricular mass was 10-fold over the ages studied, whereas total length, surface area, and volume of fibers increased approximately 3-, 8-, and 11-fold over the period. Relative rates of growth implied that fiber growth was dominated by an increase in length compared to other dimensions. The total number of myocyte nuclei ( approximately 30 x 10(6)) did not change between 5 and 25 days of age, but then almost doubled in 125-day-old rats. The number of non-myocyte nuclei increased 9-fold over the period studied in an exponential manner. The mean myocyte nucleus volume tripled between the ages of 5 and 25 days and then remained the same. The volume-weighted mean nucleus volume was highly variable and showed no significant trend with age. Our results provide support for the claim made by some researchers that myocyte proliferation had ceased by day 5 after birth, but do not provide evidence for binucleation of myocytes between 5 and 25 days after birth. Reported numbers of myocyte nuclei express a net growth and do not rule out both myocyte death and creation throughout the early postnatal period. We clearly detect an increase in the number of myocyte nuclei from 25 to 125 days but are not able to state whether this increase reflects myocyte proliferation or myocyte binucleation. To determine this would require special double-staining techniques to delineate intercalated discs and myocyte membranes. Our results agree very well with the limited data available from other studies that have used unbiased stereology. It is important to use unbiased designs that consider anisotropy as well as heterogeneity of the heart in sampling schemes, and to determine absolute quantities to avoid problems associated with interpreting changes in density (the "reference trap").