The initial reaction velocities of lactate dehydrogenase in various cell types

Abstract

The initial reaction velocities (vi) of lactate dehydrogenase in hepatocytes, cardiac muscle fibres, skeletal (gastrocnemius) muscle fibres, gastric parietal cells, ductal epithelial and acinar cells of the parotid gland, and oocytes were determined, by computer-assisted image analysis, in unfixed sections of these tissues incubated at 37 degrees C on substrate-containing agarose gel films. They were found to fit the equations vi = a1 zero A (equation 1) and vi-v = a2 zero A (equation 2) reported previously for mouse hepatocytes (Nakae & Stoward, 1993a, b), where v and zero A are, respectively, the gradients (or steady-state velocities) and the intercepts on the absorbance axis of the linear regression lines of the absorbance (A) of the final reaction product on incubation times between 1 and 3 min, and a1 and a2 are constants. Both equations 1 and 2 fitted the observed vi closely for mouse (a1 = 2.7, a2 = 2.2) and human (a1 = 3.0, a2 = 1.9) hepatocytes. However, equation 2 fitted the observed vi better than equation 1 for mouse cardiac muscle fibres (a1 = 1.5), skeletal muscle fibres (a2 = 1.2), gastric parietal cells (a2 = 1.7), acinar (a2 = 1.4) and striated ductal (a2 = 2.2) epithelial cells of the parotid gland, and oocytes (a2 = 1.6). The values of vi calculated from the two equations agreed with the observed vi to within about 11%. They ranged from 105 mumole hydrogen equivalents/cm3 cell/min units in hepatocytes to 24 units in parotid acinar cells, but for other cell types they were between 46 and 61 units. These are all considerably higher than values reported previously.