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Due to lack of low-cost and convenient measurement procedures, uncontrolled changes in spontaneous physical activity (SPA) level often are insufficiently considered as a confounding factor in rodent studies. Nonetheless, alterations in SPA can significantly impact on a wide range of physiological measurements. Therefore, we developed an accurate, low-cost video tracking procedure to allow routine assessment of SPA in the home cage of experimental animals (i.e., mice) and in the absence of any distress that might cause alterations in SPA. SPA parameters acquired (movement distance, movement time, and movement speed) with the novel tracking system were identical to those simultaneously obtained with a high-end and well-validated movement-tracking device (mean error = 0.15 ± 0.07%, r = 0.99, P &lt; 0.001). To further validate the setup, we also demonstrated caffeine-induced stimulation of SPA (195% more activity compared with vehicle, P &lt; 0.01), we adequately reproduced typical SPA fluctuations inherent to day/night cycles (146 and 702% more active during nocturnal compared with diurnal cycle for Balb/c and C57BL/6J mice, respectively, P &lt; 0.001), and we confirmed previously documented SPA differences between animal strains (24% less activity in C57BL/6J mice compared with Balb/c mice, P &lt; 0.05). Taken together, we provide data to prove that this novel low-cost methodology can be conveniently used in any mouse experiment where uncontrolled changes in SPA due to experimental interventions might confound data interpretation. By analogy, the system can be used to document a beneficial impact of therapeutic interventions on SPA in any disease mouse model.    NEW &amp; NOTEWORTHY We developed a low-cost procedure to routinely measure SPA in mice. The procedure maintains normal SPA because the animals continue to stay in their home cage in the absence of any external manipulation by the investigators and under habitual dark/light ambient conditions. This novel methodology can be conveniently used in any mouse experiment to quantify experimentally induced alterations in SPA or to assess natural variations in SPA that might confound data interpretation.

A noninterfering system to measure in-cage spontaneous physical activity in mice