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A new method to measure intestinal activity of P-glycoprotein in avian and mammalian species

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Abstract

Permeability-glycoprotein (Pgp) actively exports numerous potentially toxic compounds once they diffuse into the cell membrane of intestinal epithelial cells. We adapted the everted sleeve technique to make the first measures of intestinal Pgp function in an avian species (chicken) and in wild mammalian species, and compared them to laboratory rats. Tissues maintained both structural and functional integrity, and our method offers advantages over other in vitro techniques by using smaller intestinal sections (1 cm), and shorter incubation times (8–12 min). To determine Pgp function, we compared accumulation of [3H]-digoxin in sleeves incubated in Ringer solution with and without a transport-saturating concentration of a competitive inhibitor, cyclosporin A. We demonstrated significant variation in Pgp activity within individuals along the intestine, between populations fed different diets, and between species (laboratory rats had one-third to one-fifth the Pgp activity of wild rodents). In chicken, we also tested the effect of natural metabolites on digoxin accumulation. We found that among flavonoids, genistein (200 μM), found in soy and other legumes, but not quercetin (10, 30, 100, 330 μM) or the 3-β-glycoside isoquercetrin (100 μM), significantly increased digoxin accumulation. Among fungal metabolites, sterigmatocystin (5 μM), but not aflatoxin B1 (5 μM), significantly increased digoxin accumulation.

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Abbreviations

ABC:

ATP-binding cassette

AFB1:

Aflatoxin-B1

CsA:

Cyclosporin A

C.V.:

Coefficient of variation

DMSO:

Dimethyl sulfoxide

GEN:

Genistein

ISOQ:

Isoquercetrin

MDR:

Multi-drug resistance

PEG:

Poly-ethylene glycol

Pgp:

Permeability-glycoprotein

QUERC:

Quercetin

SGLT-1:

Sodium-dependent glucose transporter

ST:

Sterigmatocystin

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Acknowledgements

We would like to thank B. Darken for extensive help in the lab. Support for this research came from USDA (Hatch) WISO4322, NSF IBN-9723793 and IBN-0216709 to W.H.K. A.K.G was supported by an NSF pre-doctoral fellowship and a UW-Madison graduate fellowship. All research conformed to UW-Madison IACUC protocols.

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Authors and Affiliations

  1. Department of Wildlife Ecology, University of Wisconsin-Madison, 1630 Linden Dr., Madison, WI, 53706, USA

    Adam K. Green & William H. Karasov

  2. Department of Animal Sciences, University of Wisconsin-Madison, Madison, WI, 53706, USA

    David M. Barnes

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  1. Adam K. Green
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  2. David M. Barnes
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  3. William H. Karasov
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Correspondence to William H. Karasov.

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Communicated by I.D. Hume

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Green, A.K., Barnes, D.M. & Karasov, W.H. A new method to measure intestinal activity of P-glycoprotein in avian and mammalian species. J Comp Physiol B 175, 57–66 (2005). https://doi.org/10.1007/s00360-004-0462-0

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