Protein synthesis in different populations of rat hepatocytes separated according to density

Hepatocytes were isolated from fasted rats by a two-step CA++-free/collagenase perfusion method. The cells were subjected to centrifugation under mild conditions at 12 degrees C in a linear metrizamide gradient (1.075-1.12 gm/cm3). The cells were distributed in the gradient in a bell-shaped manner. According to their position in the gradient the cells were divided in five different populations. The heaviest population was omitted from the subsequent evaluation because it contained a high proportion of dead cells. The activity of alanine aminotransferase increased with increasing cell density indicating that the lightest cell population was enriched in perivenous cells, whereas the heaviest cell population had an excess of periportal cells. Protein synthesis was more rapid in the light (perivenous) cell population than in the heavy (periportal) cell population as measured by means of incorporation of radioactively labeled valine into protein. The distribution measured in vitro indicated approximately 80% higher rates in perivenous cells. On the other hand, the synthesis and secretion of export proteins were similar in all cell populations regardless of their density. Protein degradation measured as appearance of free valine in cell media was higher in the light (perivenous) cell population than in the other populations. Thus protein metabolism seemed to be faster in the light cell population.