Central corneal thickness (CCT) can be a promising source of glaucoma monitoring and diagnosis. This study evaluated changes in CCT according to experimental adjustment of intraocular pressure (IOP) in canine eyes. To adjust and measure IOP, each eye was cannulated with two 26-gauge needles under inhalant anesthesia. One needle was connected to a pressure transducer, and the other was connected to an adjustable bag of physiologic saline. IOP was stepwise increased from 10 mmHg to 70 mmHg in 10 mmHg increments (Group T). IOP was maintained at 15 mmHg (Group C15), 30 mmHg (Group C30), 45 mmHg (Group C45), 60 mmHg (Group C60) and 75 mmHg (Group C75) during the experiment. CCT was measured with an ultrasonic pachymeter every 10 min after cannulation. There was a significant difference in the effect of time on CCT (P<0.001) and difference in CCT (dCCT; P<0.001) between groups. The CCT of group C15 remained constant during the experiment. However, group T showed an initial decrease and then an increase after passing the lowest point. Group C30 showed decreasing values for 30 min, after which the values remained constant. The values in Group C45 showed no changes for 40 min and then increased. The values in group C60 showed no change for 20 min and then increased. Group C75 showed a steady increase. In conclusion, the CCT showed two core changes according to increased IOP. This study provides essential basic data to enable further investigation into the association of IOP and CCT in dogs.