Ability to image cerebral metabolic rate of oxygen (CMRO(2)) is essential for studying the fundamental role of oxidative metabolism in brain function and disease. We have demonstrated recently that three-dimensional (3D) CMRO(2) images can be obtained in the rat brain during a 2-min (17)O(2) inhalation using the (17)O MR spectroscopic imaging (MRSI) approach at high field. The feasibility for establishing a completely noninvasive approach for imaging CMRO(2) has also been demonstrated. In this study, we further explored the feasibility of (17)O MRSI approach for performing repeated CMRO(2) measurements within a short period of time and evaluated the reproducibility of the repeated measurements. Subsequently, we applied the (17)O MRSI approach to measure CMRO(2) and cerebral blood flow (CBF) values at two brain temperatures in the alpha-chloralose anesthetized rat brain at 9.4 T. Finally, we tested the validity of simplified model for noninvasively determining CMRO(2) in normothermic and hypothermic rat brain. The results show (i) an excellent reproducibility among repeated measurements of 3D CMRO(2) images under the same physiologic condition; (ii) a 44% decrease of CMRO(2) across the rat brain at mild hypothermic (32 degrees C) condition as compared with normothermic (37 degrees C) condition; and (iii) a close correlation between CMRO(2) and CBF within a relatively wide physiologic range. This study demonstrates the capability of (17)O MRSI approach for noninvasively imaging CMRO(2) and its changes caused by physiologic perturbation. This approach, thus, should provide a promising neuroimaging modality for studying oxidative metabolism and bioenergetics associated with brain functions and diseases.