In an attempt to establish the functional organization of the hepatic parenchymal unit, we used histo- and microchemical procedures to assess metabolic liver cell heterogeneity at the level of the primary lobule. Because of the close interrelation of glucogenesis and ketone body formation, and in view of the distinct regional differences of the in vivo activity of glucose-6-phosphatase (G6Pase), these techniques were used on livers from male rats to investigate the distribution of the ketogenic enzyme, 3-hydroxybutyrate dehydrogenase (3-HBDH), during the post-resorptive phase. A close reciprocity was found between the general increase in the activity of 3-HBDH and the decrease of the in vivo activity of G6Pase along the sinusoidal axis, and also with regard to enzyme gradients along sinusoids of different origin. The activity of the ketogenic enzyme was higher throughout septal than portal sinusoids, whereas the opposite applied to the glucogenic enzyme. Histo- and microchemical data support the concept of a lobular parenchymal unit composed of "primary lobules," and show also that hepatocyte function varies with cell location along the sinusoidal axis and with the origin of the sinusoids.