Autosomal dominant polycystic kidney disease (ADPKD) is a progressive kidney disease where the size of the kidneys is correlated to the stage of kidney failure. Total kidney volume (TKV) is used as a prognostic marker to determine disease stage, progression, and possible effect of treatment. It has been shown that water restriction is associated with reduced kidney volume in healthy subjects. The aim of this study was to evaluate the relationship between TKV and hydration status in ADPKD patients. 40 ADPKD patients with chronic kidney disease stage 1 - 3 were randomized to either 3 hours of water restriction (n = 21) or 1 hour of water loading (n = 19; intake: 20 mL/kg). The patients had a mean age of 38 years (19 - 73) and mean plasma creatinine level of 91 (54 - 178) µmol/L. Magnetic resonance imaging of the kidneys was performed before and after intervention, and TKV was measured using the ellipsoid formula. Water restriction resulted in an insignificant 0.67% increase in TKV (median: 1.48, interquartile range (IQR): 6.1, range: -1. - 4.5). Water loading resulted in an insignificant 2.67% increase in TKV (median: 3.18, IQR: 11.4, range: -3.6 - 7.8). Interestingly, a 7.09% increase in right-kidney volume was found after water loading (median: 5.58, IQR: 9.4 range: 1.9 - 11.3, p < 0.05), whereas the left-kidney volume showed an insignificant decrease of 0.18% after water loading (median: -1.65, IQR: 18.0, range: -12.5 - 5.5). We found in ADPKD patients that neither short-term water restriction nor acute water loading had significant effects, suggesting that the use of TKV for disease staging is independent of hydration level in these patients.