The effect of acute in vivo epidermal growth factor (EGF) exposure on intestinal absorptive surface area was examined in blind jejunal loops in New Zealand White rabbits (500-900 g). Brush-border surface area was assessed by electron microscopy at 30 and 120 min, and total surface area was assessed by the fluorophore 1-[4-(trimethylamino)phenyl)]-6-phenylhexatriene (TMA-DPH) at 10 and 60 min after EGF exposure. Mucosa obtained at 120 min was also used for morphometrics and analysis of membrane composition and fluidity. Brush-border surface area was significantly increased in EGF-treated tissue after 30 (42%) and 120 min (60%). Total absorptive surface area measured by TMA-DPH was increased more than twofold after 10 and 60 min EGF exposure. The increase in absorptive surface area was abolished by pretreatment with intravenous verapamil. Mucosal morphometrics and membrane sucrase activity, total phospholipids, cholesterol content, cholesterol-to-phospholipid ratio, phospholipid species (wt%), phospholipid fatty acid composition, and fluidity did not differ between control and EGF-treated tissue. These findings indicate that EGF can rapidly increase brush-border surface area by a mechanism that appears to be due to a redistribution of existing performed microvillus plasma membrane.