Key words: functional magnetic resonance imaging; relative cerebral blood volume; bicuculline; pharmacological stimulation; mouse; iron-oxide nanoparticles; transcutaneous blood gas monitoring Genetic engineering and transgenic technology have led to advances in the generation of animal models that develop aspects of various brain pathologies, and have emphasized the need to develop accurate methods when phenotyping these animals. Such models would benefit from the development of noninvasive methods to study disease progression while allowing for a correlation with behavioral studies. The purpose of the present study was to assess the feasibility of functional magnetic resonance imaging (fMRI) in mice by monitoring changes in local relative cerebral blood volume (CBV rel ) during pharmacological stimulation under controlled physiological conditions, and hence to assess the potential of pharmacological fMRI as a noninvasive method to characterize transgenic and knock-out mice models. fMRI has proven its utility for the noninvasive mapping of brain function with high temporal and spatial resolution. Depending on the method applied, image contrast in fMRI is determined either by the oxygenation state of hemoglobin (BOLD) (1) or by changes in cerebral hemodynamics, i.e., cerebral blood flow (CBF) (2,3) and CBV (4). Using superparamagnetic iron-oxide nanoparticles as a blood-pool contrast agent, a local CBV rel increase is reflected by an increase in the amount of magnetite nanoparticles in the activated tissue and, correspondingly, by a decrease in the signal intensities in T 2 -weighted images. High-resolution CBV rel maps can be derived from the signal attenuation once the tracer has reached a steady-state blood concentration. Applying this CBV rel -fMRI method, activation in the rat sensory cortex caused by electrical stimulation of the forepaw was demonstrated (5,6). Hemodynamic changes associated with a pharmacological stimulation paradigm have been demonstrated in rats, and could be used to study drugs at the specific receptor level. Systemic infusion of the GABA A antagonist bicuculline led to an alteration in neuronal activity in cortical and subcortical brain structures. Dose-dependent CBV rel increases have been measured in various brain structures (e.g., the cortex, thalamus, and caudate putamen) in the rat (7).It is well known that physiological parameters, such as body temperature and blood gases, will affect CBV values. Mandeville et al. (6) reported CBV rel increases on the order of 50% in rats during hypercarbia by increasing arterial CO 2 tension (PaCO 2 ) from 25-55 mmHg. Although reports on the quantitative relationship between changes in CBV rel and PaCO 2 differ considerably, it is obvious that control and continuous monitoring of blood CO 2 and blood oxygenation are critical for detecting small alterations of cerebrovascular parameters in the mouse. Although larger animals can sustain frequent blood samplings, noninvasive monitoring of blood gases has been found to be essential when studying m...