Volume 115, Issue 5, Pages 1172-1178 (November 1998) Molecular analysis of the adaptive response of intestinal bile acid transport after ileal resection in the rat  Christopher P. Coppola*, John R. Gosche*, Marco Arrese‡, Bret Ancowitz§, Jill Madsen∥, Jon Vanderhoof¶, Benjamin L. Shneider#  Gastroenterology  Volume 115, Issue 5, Pages 1172-1178 (November 1998) DOI: 10.1016/S0016-5085(98)70088-5 Copyright © 1998 American Gastroenterological Association Terms and Conditions

Fig. 1 Schematic diagram of surgical procedures in the rat. Each bar represents the 100 cm of rat small intestine from duodenum to cecum. Black areas represent resections. Numbers and vertical single black lines delineate segments of intestine harvested for analysis 3 weeks after surgery. Vertical double black lines represent sites of transection and reanastomosis. Gastroenterology 1998 115, 1172-1178DOI: (10.1016/S0016-5085(98)70088-5) Copyright © 1998 American Gastroenterological Association Terms and Conditions

Fig. 2 Linear range of ASBT Western blotting. Graphic analysis of the relationship between ASBT phosphorimager signal intensity (y axis) and the quantity of crude ileal BBMVs loaded (x axis). The best fit equation for either a linear or logarithmic relationship is shown. A linear relationship between ASBT signal and membrane quantity can be seen if between 5 and 20 μg of ileal membranes are loaded. Signal intensity appears to plateau with ≥30 μg of BBMV. The actual blot can be seen in the inset. Gastroenterology 1998 115, 1172-1178DOI: (10.1016/S0016-5085(98)70088-5) Copyright © 1998 American Gastroenterological Association Terms and Conditions

Fig. 3 Expression of ASBT, ILBP, and bile acid transport capacity along the longitudinal axis of the gastrointestinal tract. Functional and molecular analyses of the expression of ASBT and ILBP are shown. (A) Deciles of the small intestine length have been analyzed for ASBT mRNA and protein quantitation along with sodium-dependent bile acid transport activity. The values are the mean of 3 separate determinations. Overlapping expression of these proteins and transport activity are detected in the terminal 30% of the small intestine. (B) Representative actual Northern and Western blots of ASBT and ILBP using samples from the segments indicated on the x axis in A. Gastroenterology 1998 115, 1172-1178DOI: (10.1016/S0016-5085(98)70088-5) Copyright © 1998 American Gastroenterological Association Terms and Conditions

Fig. 4 Representative Western blot of sham-operated vs. ileal resection. Three separate samples of either ileal membranes (ASBT) or homogenates (ILBP) from either sham-operated animals (S) or surgically resected animals (R) have been blotted as described in Materials and Methods for their respective proteins. No difference in the abundance of either protein is observed in sham vs. surgically resected animals. Gastroenterology 1998 115, 1172-1178DOI: (10.1016/S0016-5085(98)70088-5) Copyright © 1998 American Gastroenterological Association Terms and Conditions

Fig. 5 Representative Western blot of ASBT and ILBP expression in intestinal segments after distal ileal resection. BBMV or homogenates of segments 1–3 (S1, S2, and S3) of the sham-operated animals (see Figure 1) and segments 1 and 3 (R1 and R3) of the surgically resected animals have been blotted for ASBT and ILBP protein expression. No expression of ASBT or ILBP can be seen in segment 1 of either the sham or resected animals. kDa, kilodaltons. Gastroenterology 1998 115, 1172-1178DOI: (10.1016/S0016-5085(98)70088-5) Copyright © 1998 American Gastroenterological Association Terms and Conditions

Fig. 6 Western blot analysis of the effect of bile acid feeding on the expression of ASBT and ILBP. Four separate samples of either ileal membranes (ASBT) or homogenates (ILBP) from either sham-fed animals (C) or 0.5% taurocholate-fed animals (TC) have been blotted as described in Materials and Methods for their respective proteins. No difference in the abundance of either protein can be observed in sham- vs. taurocholate-fed animals. Gastroenterology 1998 115, 1172-1178DOI: (10.1016/S0016-5085(98)70088-5) Copyright © 1998 American Gastroenterological Association Terms and Conditions