We measured the arterial Po2 and AaDo2 in open-chest dogs respired with air and with 5% CO2 in air at various lung volumes using a constant hyperventilatory pattern. The AaDo2 was an inverse function of lung volume with both air and 5% CO2 and was also an inverse function of both alveolar and arterial Pco2 values except at quite high lung volumes. There were two series of closed-chest experiments. In the first series, ventilation was varied to produce alveolar Pco2 and Po2 changes. In the second series, the dogs were hyperventilated at a constant rate and Pco2 was varied by adding CO2 with alveolar Po2 levels kept relatively constant. In both series the AaDo2 was inversely related to Pco2. We conclude that, in dogs, the AaDo2 is independent upon the Pco2 and speculate that this may be related to the effect of CO2 on collateral ventilation, although the Bohr effect may account for some of the dependence.