The aim of this study was to test whether immobilization and intravenous volume load as stressors influence the intraocular pressure in rabbits. Rabbits were immobilized for 1 h in a horizontally placed plastic tube with an internal diameter of 13.2 cm and a length of 33.2 cm. After immobilization, rabbits received rapid intravenous drip infusion of 5% glucose solution, 20 ml/kg of body weight, in 5 min. The intraocular pressure immediately after immobilization (11.2 ± 3.0 mm Hg; mean and standard deviation) was significantly higher compared to control rabbits without immobilization (9.2 ± 1.0 mm Hg, Student’s t test, p = 0.0462). This difference became significantly larger when volume load was exerted on both groups of rabbits (22.7 ± 5.6 versus 16.4 ± 2.2 mm Hg, p = 0.0067). Serum levels of cortisol, adrenaline, and noradrenaline were significantly elevated after immobilization (p = 0.0002, p = 0.0271, p = 0.0296, respectively). Venous pressure of the ear tended to increase in rabbits immediately after immobilization (15.8 ± 3.1 mm Hg) compared with control rabbits (8.5 ± 2.3 mm Hg), and the difference became significant when volume load was exerted on both groups of rabbits (20.8 ± 7.4 versus 9.2 ± 4.8 mm Hg, p = 0.0211). In conclusion, we clearly demonstrated that physical stress due to immobilization, especially in combination with volume load, increased intraocular pressure in rabbits.