Energy-dependent accumulation of glycine and glycyl-L-valine within the small intestinal mucosa in a chicken model of in vitro local oxygenation of the small intestinal preparation was studied. It has been shown that the most effective bilateral oxygenation significantly increase accumulation of glycyl-L-valine in the proximal segment as compared to that under oxygenation only from serosal surface both in the fed and 24-hour fasted chickens, whereas in other segments these differences was less apparent. This may be due to increased H+/ peptide cotransporter expression in the proximal segment. Thus the bilateral oxygenation probably may turn on an additional amount of already existing (but non-functional during serosal oxygenation) H+/ peptide co-transporters. Moreover, low glycine transporter expression may be the reason why supplemental oxygen (bilateral oxygenation) has no effect on glycine accumulation in the distal segment of fed chickens. A 48-hour fasting decreases glycyl-L-valine accumulation in the proximal (and medial) segments, possibly due to progressive decrease in villus height. It is concluded that: a) the accumulation rate of glycine was greater when presented as the glycyl-L-valine than when presented as the equivalent amount of free amino acid; b) the rates of accumulation of glycyl-L-valine are highest in the proximal segment, decrease in the medial segment and are the lowest in the distal segment; c) the serosal oxygenation is less effective than the mucosal and bilateral oxygenation, which markedly stimulates accumulation of nutrients in the intestinal mucosa; d) a 24-hour fasting increases glycyl-L-valine accumulation in the proximal segment only, while glycine uptake was increased in the distal segment.