An isolated vascularly perfused rat intestine system was utilized to examine various aspects of zinc absorption in an attempt to more clearly examine the mechanisms involved. The lumen was perfused with a modified tissue culture medium containing 65Zn. The vascular system was perfused from the superior mesenteric artery to the portal vein, with Krebs-Ringer bicarbonate buffer containing 5% rat serum. The criterion for absorption was the amount of radioactivity transferred to the vascular perfusate. When the intestines were obtained from rats that had consumed a zinc-deficient diet the amount of zinc absorbed increased markedly. Conversely, elevation of zinc status decreased the amount of 65Zn that could be transferred to the vascular perfusate. These data strongly suggest that the isolated, perfused rat intestine retains the ability to exercise homeostatic control over 65Zn absorption. Transfer of infused 65Zn to the vascular perfusate was significantly decreased by aspirin, phytate, and prostaglandin E2. Uptake of 65Zn from the lumen into the intestinal cells was significantly increased by histidine and significantly decreased by phytate and prostaglandin E2. Thus, the isolated, vascularly perfused rat intestine appears to be capable of differentiating between the cellular uptake and cell to plasma transfer phases of zinc absorption.