CO2 kinetics were studied under conditions in which CO2 partial pressure and either the bicarbonate or hydrogen ion concentrations were unequal in perfusate and bath. A glass pH microelectrode and microcalorimeter were used to measure pH and total CO2 in perfused and collected fluids. Luminal appearance or disappearance of CO2 was determined from the change in concentrations of CO2 and acidic moieties of the buffers between perfused and collected fluids. The pH was not measurably affected by the tubule processes of metabolism, hydrogen ion secretion, and buffer transport because of relatively high flow rates. Since CO2 appearance and disappearance were directly related to CO2 partial pressure differences across the tubule (r = 0.94), this technique provides a valid estimate of transepithelial CO2 flux in response to the driving force of a CO2 partial pressure difference. From flux per unit of driving force and from estimates of resistances to CO2 diffusion in both internal and external unstirred layers, we obtained a transepithelial CO2 permeability of approximately 10(-4) cm3 . s-1 . cm tubule length-1. This corresponds to a diffusion coefficient through the tubule epithelium about half that through an equivalent thickness of water. We conclude that rabbit proximal convoluted tubules are so highly permeable to CO2 that even if all the filtered bicarbonate were reabsorbed by the generation of CO2 in the lumen, the tubular fluid CO2 partial pressure would exceed that of the peritubular blood by less than 4 mmHg.