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. 1999 Feb;44(2):218–225. doi: 10.1136/gut.44.2.218

Macromolecular transport across the rabbit proximal and distal colon

J Hardin 1, M Kimm 1, M Wirasinghe 1, D Gall 1
PMCID: PMC1727367  PMID: 9895381

Abstract

Background—Although many studies have investigated macromolecular uptake in the stomach and small intestine, little is known about macromolecular uptake in the colon. 
Aims—To investigate the mechanisms involved in the transport of large antigenically intact macromolecules across the proximal and distal colonic epithelium in the rabbit. 
Methods—The mucosal to serosal movement of bovine serum albumin (BSA) was examined in modified Ussing chambers under short circuited conditions. The mucosal surface was exposed to varying concentrations of BSA, and after a 50 minute equilibration period, the mucosal to serosal flux of immunologically intact BSA was determined by ELISA. Total BSA flux was determined by the transport of radiolabelled 125I-BSA. 
Results—Intact BSA transport in proximal and distal colonic tissue showed saturable kinetics. Intact BSA transport in the proximal and distal segment was 7% and 2% of the total 125I-BSA flux respectively. Immunologically intact BSA transport in the distal segment was significantly less than that in the proximal segment. Intact BSA transport in the proximal colon was significantly reduced following treatment with sodium fluoride, colchicine, and tetrodotoxin. Cholinergic blockade had no effect on the uptake of intact BSA. 
Conclusion—The findings indicate that the transport of intact macromolecules across the proximal and distal large intestine is a saturable process. Further, intact BSA transport in the proximal colon is an energy dependent process that utilises microtubules and is regulated by the enteric nervous system. 



Keywords: colon; transport; protein

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Figure 1 .

Figure 1

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Representative micrographs of stripped colonic tissue obtained 80 minutes after mounting in the Ussing chamber. (A) proximal colon; (B) distal colon. Original magnification × 400. 


Figure 2 .

Figure 2

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Dose response curve for intact BSA transport in the proximal colon. Data represent the mean (SEM) of at least three tissues at each concentration. 


Figure 3 .

Figure 3

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Dose response curve for intact BSA transport in the distal colon. Data represent the mean (SEM) of at least three tissues at each concentration. 


Figure 4 .

Figure 4

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Metabolic inhibition and neural regulation of intact BSA transport in proximal colonic tissue. NaF (n=6), colchicine (n=10), TTX (n=9), ATR + HEX (n=4), controls (n=the number in each experimental group). All measurements were performed in paired tissue. *p<0.05. 


Figure 5 .

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Metabolic, microtubule, and neural inhibition of total (intact and degraded) BSA. n=4 for all groups. All measurements were performed in paired tissue. 


Figure 6 .

Figure 6

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Immunoblot of BSA collected (A) fresh (immediately after preparation), and (B) from the serosal chamber after 80 minutes. Immunologically intact BSA is visualised as a single 66.2 kDa band in the sample obtained from (B). 


Figure 7 .

Figure 7

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Autoradiograph of mucosal solution obtained at 80 minutes containing 125I-BSA (A) and freshly prepared 125I-BSA (B). BSA was not degraded prior to transport across the tissue and remained conjugated to 125I. 


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