Ventilation in larger animals and humans is gravity dependent and mainly distributed to the dependent lung. Little is known of the effect of gravity on ventilation distribution in small animals such as rodents. The aim of this study was to investigate gravity-dependent ventilation distribution and regional filling characteristics in rats. Ventilation distribution and regional lung filling were measured in six rats using electrical impedance tomography (EIT). Measurements were performed in four body positions (supine, prone, left and right lateral), and all animals were ventilated with increasing tidal volumes from 3 to 8 mL kg(-1). The effect of gravity on regional ventilation distribution was assessed with profiles of relative impedance change and calculation of the geometric centre. Regional filling was measured by calculating the slope of the plot of regional versus global relative impedance change on a breath-by-breath basis. Ventilation was significantly distributed to the non-dependent lung regardless of body position and tidal volume used. The geometric centre was located in the dependent lung in all but prone position. The regional filling characteristics followed an anatomical pattern with the posterior and the right lung generally filling faster. Gravity had little impact on regional filling. Ventilation distribution in rats is gravity dependent, whereas regional filling characteristics are dependent on anatomy.