1 Department of Gastroenterology, University of Heidelberg, 69115 Heidelberg, Germany; and 2 Graduate School of Pharmaceutical Sciences, The University of Tokyo, 113 - 0033 Tokyo, Japan
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ABSTRACT |
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Multidrug
resistance-associated protein 3 (MRP3; symbol ABCC3), has been shown to
mediate ATP-dependent transport of organic anions including
17-glucuronosyl estradiol, glucuronosyl bilirubin, monovalent, and
sulfated bile salts. MRP3 mRNA expression was reported in
rat intestine suggesting a role of MRP3 in the intestinal transport. We
examined the expression and localization of MRP3 in rat small and large
intestine by RT-PCR, immunofluorescence, and immunoblot
analysis. MRP3 was identified in all intestinal segments by
RT-PCR. MRP3 expression was low in duodenum and jejunum but markedly
increased in ileum and colon. With the use of a rat MRP3 specific
antibody, MRP3 was localized to the basolateral domains of enterocytes.
Immunofluorescence analysis and immunoblot analysis confirmed a strong
expression of rat MRP3 in ileum and colon. In contrast, MRP2 was
predominantly expressed in the proximal segments of rat small
intestine. Our findings demonstrate a high expression of rat MRP3 in
ileum and colon and provide evidence for an involvement of MRP3 in the
ATP-dependent transport of organic anions, including bile salts from
the enterocyte into blood.
enterohepatic circulation; intestinal transport; organic anions; bile acids
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INTRODUCTION |
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MEMBERS OF THE MULTIDRUG RESISTANCE-associated protein (MRP) family have been identified to mediate ATP-dependent transport of organic anions across membranes. MRP substrates include xenobiotic and endogenous lipophilic substances conjugated with glutathione, glucuronate, and sulfate (3, 22). Nine MRP isoforms (ABCC1-6 and ABCC10-12) have been identified in humans and have been shown to exhibit a tissue-specific distribution (19, 26, 27, 39). In humans, MRP1, MRP2, MRP3, and MRP5 are expressed to different extents in small and large intestine, as revealed by RNAse protection assay and Northern blot analysis (1, 26, 31). Differential expression of MRP1 and MRP2 proteins in the intestine has been studied in more detail in rodents (30, 31, 33). MRP1 expression in the small intestine was low and was shown to be limited to undifferentiated enterocytes at the base of the crypts (33). The apical MRP-isoform MRP2 was predominantly expressed in the proximal part of rat small intestine (11, 31) and has been suggested to contribute to organic anion excretion from the enterocyte into the lumen of the gut (11, 30). Thus in rat, neither MRP1 nor MRP2 can account for the well-recognized basolateral transport of endogenous and xenobiotic substances including hormones, bile salts, and drugs from the small intestine into blood.
Recently, MRP3 has been cloned from rat and human liver
(14, 23) and was subsequently localized to the basolateral
membrane of hepatocytes (25, 28), cholangiocytes
(28, 37), and MRP3-tranfected polarized cells (24,
28). MRP3 substrates include 17-glucuronosyl estradiol
(E217
G) (15, 44), glucuronosyl bilirubin
(22), monovalent bile salts (i.e., taurocholate and glycocholate) (16, 44), and sulfated bile salts (i.e.,
taurochenodeoxycholate-3-sulfate, taurolithocholate-3-sulfate)
(16). In addition to liver and kidney, high
MRP3 mRNA expression was found in rat and human intestinal tissues (14, 23, 26), suggesting a possible role of MRP3 in the intestinal secretion of organic anions (13,
14).
To further characterize the role of MRP3 in the intestine, we examined the expression and localization of rat MRP3 in duodenum, jejunum, ileum, and colon using RT-PCR, immunofluorescence microscopy, and immunoblot analysis. Our results demonstrate that MRP3 is localized to the basolateral domain of rat enterocytes and is expressed at high levels in the ileum and colon of rat intestine. MRP3 may, therefore, represent a major export pump for organic anions in the rat distal intestine and may be involved in the enterohepatic circulation, mediating the ATP-dependent secretion of bile salts, steroids, and drugs from the enterocytes into blood.
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MATERIALS AND METHODS |
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Rat tissue samples. Adult male Wistar rats weighing 180-220 g were purchased from Charles River Wiga (Sulzfeld, Germany) and were kept according to the Guide for the Care and Use of Laboratory Animals (NIH publication 86-23, revised 1985). Rats were anesthetized with Rompun (15 mg/kg; Bayer, Leverkusen, Germany) and Ketanest (100 mg/kg; Parke-Davis, Morris Plains, NJ). Segments (5-cm long) of duodenum, jejunum, ileum, proximal colon, and distal colon were rinsed with ice-cold PBS and opened lengthwise, and mucosa was obtained by scraping as described (6). Mucosa samples were frozen in liquid nitrogen until use.
Antibodies. Rat MRP2-specific EAG15 antibody was raised in rabbit against a synthetic peptide containing the 15 COOH-terminal amino acids of the deduced rat MRP2 sequence (4). The polyclonal MRP3 antibody was obtained by immunizing rabbits with a maltose-binding fusion protein containing the 136 amino acids corresponding to 838-973 of the deduced rat MRP3 amino acid sequence (32, 37). The mouse monoclonal antibody C219 was purchased from Centocor (Malvern, PA) and reacts with rat P-glycoproteins including MDR1a, MDR1b, and MDR2. Cy2-conjugated goat anti-rabbit antibody and Cy3-conjugated goat anti-mouse antibody were purchased from Dianova (Hamburg, Germany).
RNA-isolation, RT-PCR, and sequencing.
Total RNA was isolated from mucosa samples using the RNAClean solution
(Hybaid-AGS, Heidelberg, Germany) according to the manufacturer's
manual. RT-PCR was performed with Ready-To-Go RT-PCR Beads (Amersham
Pharmacia Biotech) according to the manufacturer's manual. The RNA was
reverse transcribed with pd(T)12-18-primer using 2 µg total RNA at 42°C for 30 min. For PCR, 35 cycles of denaturation
(94°C for 60 s), annealing (52°C for 60 s), and
elongation (72°C for 90 s) were performed using the following
primer pairs selective for the respective MRP family member:
okoplus3 (5'-GGGATAAATCTCAGTGGT-3') against okorev2
(5'-ATATGCTCCACAGAGTTG-3') (NIH/GenBank accession no. X96,393) for rat
MRP2 and omrp3rat.for1 (5'-TAAGGTGGATAGCAACCAG-3') against
omrp3rat.rev1 (5'-GGCTAGGCACACGAGCT-3') (NIH/GenBank accession no.
AB010467) for rat MRP3. The -actin control PCR was
performed using a commercially available
-actin primer mixture
(Stratagene, Amsterdam, The Netherlands). Amplified fragments were
analyzed by agarose gel electrophoresis, purified with GFX PCR DNA and Gel Band Purification Kit (Amersham Pharmacia Biotech) and sequenced by
the dideoxynucleotide chain termination method of Sanger using the PE
DNA-Sequencing Kit (Perkin-Elmer Applied Biosystems, Warrington, United Kingdom).
Immunoblot analysis.
Mucosa samples were homogenized on ice with PBS in the presence of
protease inhibitors (Protease inhibitor cocktail; Sigma, Deisenhofen,
Germany) using a glass homogenator (Wheaton, Millville, NJ).
Samples obtained from different rat intestinal segments (50 µg
protein) or rat liver homogenate (30 µg protein) were mixed with
sodium dodecyl sulfate sample loading buffer, incubated at 37°C for
60 min, and separated on 7.5% polyacrylamide gels in the presence of
-mercaptoethanol (29). After transfer to polyvinylidene difluoride membranes, blots were blocked for 1 h in 5% low fat dried milk dissolved in Tris-buffered saline containing 0.05% Tween 20 (TTBS). Membranes were incubated for 1 h with the primary and
secondary antibody, respectively, diluted in 5% dried milk dissolved
in TTBS. After each incubation, blots were washed three times with
TTBS. Antibody binding was detected using horseradish peroxidase-conjugated secondary antibodies (Bio-Rad,
Muenchen, Germany) and the enhanced chemiluminescence technique
(Amersham-Buchler, Braunschweig, Germany). The following dilutions of
antibodies were used: EAG15 at 1:5,000; anti-MRP3 antibody at 1:500;
horseradish peroxidase-conjugated goat anti-rabbit antibody at 1:2,500.
Immunoreactive bands on autoradiography films were scanned (Epson GT
9600; Epson, Tokyo, Japan) and quantified using Raytest image software
(Raytest, Straubenhardt, Germany).
Immunofluorescence microscopy.
For immunofluorescence microscopy, rat intestinal tissue was mounted in
Tissue-Tek (Miles, Elkhart, IN) and deep-frozen in liquid nitrogen.
Tissue sections (2-4 µm) were prepared with a cryotome (model
2800E FrigoCut; Leica, Nussloch, Germany), air-dried for 2 h, and
fixed for 10 min with 3% paraformaldehyde/PBS followed by cold
methanol (20°C) for 10 s. After rehydration with PBS, sections
were incubated with the primary antibodies for 60 min and then washed
three times for 10 min with PBS and incubated with the combined
secondary antibodies for 30 min. After being washed three times with
PBS, sections were rinsed with distilled water, air-dried, and mounted
with Moviol (Hoechst, Frankfurt, Germany). All antibodies were diluted
in PBS containing 5% fetal calf serum and 0.05% Tween at the
following dilutions: EAG15 at 1:100; anti-MRP3 antibody at 1:100; C219
at 1:10; Cy2-conjugated anti-rabbit IgG at 1:200; and Cy3-conjugated
anti-mouse IgG at 1:500. Nuclei were stained with Hoechst 33258 (Molecular Probes, Eugene, OR). Micrographs were taken using an Olympus
AX70 microscope on Kodak Elite II 400 ASA films (Kodak, Rochester, NY).
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RESULTS |
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RT-PCR analysis of MRP3 and MRP2 mRNA expression.
MRP3 and MRP2 mRNA expression in duodenum,
jejunum, ileum, and colon was analyzed by RT-PCR using rat-specific
primers against MRP3 and MRP2. Amplification of a
661-bp fragment of -actin was used as an internal control,
demonstrating the integrity of the isolated total RNA (Fig.
1). The MRP amplification products showed the expected size of 784 and 868 bp for rat MRP3 and
MRP2, respectively. The specificity of the PCR products was
confirmed by sequencing. MRP3 expression was low in the
proximal part of the small intestine showing a gradual increase towards
the ileum. The strongest MRP3 expression was detected in rat
colon (Fig. 1). MRP2 expression was detected in all segments
of the small intestine, with the strongest bands observed in jejunum
(Fig. 1). In contrast to MRP3, no MRP2-specific
products were obtained in rat colon.
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Localization of MRP3 to the basolateral domain rat enterocytes.
Double-label immunofluorescence microscopy was used to study
localization and distribution of MRP2 and MRP3 in the different parts
of rat intestine. MRP2 expression was restricted to the apical domains
in enterocytes of the small intestine (Fig.
2B) as described previously
(31). The C219 antibody was used to identify the apical
domains of enterocytes (Fig. 2A). Superimposition of both
images revealed colocalization of both signals yielding in a yellow
color (Fig. 2C). MRP2 expression increased from duodenum to
jejunum, where the strongest immunostaining was observed (Fig. 2B). In the terminal ileum, the immunofluorescent staining
was less intense. In rat colon, no specific apical signals were
obtained. In general, the fluorescence intensity in the small intestine decreased from the tip of the villus to the crypt, where no staining was observed (not shown).
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Differential expression of MRP2 and MRP3 in rat intestine.
Immunoblots on mucosa samples from duodenum, jejunum, ileum, and colon
were probed with polyclonal antibodies directed against rat MRP2 and
rat MRP3. Incubation with the EAG15 antibody against rat MRP2 resulted
in the specific 190-kDa band in duodenum, jejunum, and, much weaker, in
ileum of rat small intestine (Fig. 5).
Compared with jejunum (100%), where MRP2 expression was the highest,
MRP2 expression decreased to 76 ± 24 and 56 ± 16% in
duodenum and ileum, respectively. The lowest MRP2 expression was found
in colon (6 ± 1%). MRP3 expression in the different parts of rat
intestine was analyzed using a rat MRP3 specific polyclonal antibody
(32, 37). Immunoblot analysis of mucosa samples revealed a
band at 190 kDa in all segments of rat intestine. As reported
previously (32, 37), an additional band at ~120 kDa was
observed in all mucosa preparations (not shown). The intensity of this
band corresponded to the intensity of the 190-kDa band; however, it is
unclear how this band is related to the full-size transporter. In
contrast to the MRP2 distribution, the highest MRP3 expression was
found in rat ileum (100 ± 9%) and colon (91 ± 18%) as
predicted by the immunofluorescence analysis (Fig. 4), whereas MRP3
expression in duodenum (26 ± 13%) and jejunum (28 ± 9%)
was considerably low.
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DISCUSSION |
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Transport of organic anions across the enterocyte basolateral membrane into the blood represents an important step in intestinal absorption and has been studied in vivo using the intestinal loop model (10, 21, 36). Furthermore, the recycling of biliary constituents such as bile salts, steroids, vitamins, and drugs, known as enterohepatic circulation (reviewed in Ref. 5), involves reabsorption and transport across the enterocyte basolateral membrane and depends on distinct transport systems at the apical and basolateral membrane of enterocytes (38). However, the molecular identity of the transport proteins involved is largely unknown.
Transport studies using multidrug resistant cancer cell lines and transfected cell lines have identified members of the MRP-family as ATP-dependent transporters for lipophilic substances conjugated with glutathione, glucuronate, and sulfate (3, 22). Recent findings have demonstrated extensive expression of MRP3 mRNA in tissues of rat and human intestine (13, 23, 25, 26). Functional analysis indicated that glucuronate conjugates are preferred MRP3 substrates and that, in contrast to MRP1 and MRP2, glutathione conjugates are poor substrates for MRP3 (15). Moreover, rat MRP3 exhibits an unique ability to transport monovalent (taurocholate and glycocholate) and sulfated bile salts (taurolithocholate-3-sulfate and taurochenodeoxycholate-3-sulfate) (16, 44). To further characterize the role of MRP3 in intestinal transport, we studied the expression and localization of MRP3 in different segments of rat intestine using RT-PCR analysis, immunofluorescence analysis, and immunoblot analysis.
In the present study, we have localized rat MRP3 to the basolateral membrane domain of enterocytes (Figs. 2, E and F, and 3). The basolateral localization enables this conjugate export pump to contribute to the secretion of organic anions from enterocytes into blood. Moreover, our results were consistent with a high expression of MRP3 in ileum and colon, suggesting an important role for MRP3-mediated uptake in the distal part of rat intestine. MRP3 expression in duodenum and jejunum was low (Fig. 4). In contrast, MRP2 was mainly expressed in the proximal part of rat small intestine with a maximum expression in jejunum. The latter results are in good agreement with previous findings on MRP2 expression in rat intestine (11, 30, 31), indicating the reliability of the mucosa isolation technique used (6). Differential expression of MRP3 with respect to the villus crypt axis resembles MRP2 distribution in the proximal rat intestine, for which increasing protein expression has been demonstrated from the crypt to the villus (31, 40). Similarly, phase II enzyme expression increases from crypt to villus (8, 34), underlining the coordinate expression and cooperative function of conjugation enzymes with conjugate export pumps.
Considering the transport characteristics, localization, and expression
pattern of rat MRP3, the uptake of the following compounds from
intestine into blood may be, in part, attributed to ATP-dependent transport across the enterocyte basolateral membrane via MRP3. First,
bile salts, known to undergo extensive enterohepatic circulation (reviewed in Ref. 17), are mainly reabsorbed in the ileum
and have been identified as MRP3 substrates (16, 44).
Second, ethinylestradiol and other steroids are efficiently
glucuronidated in the intestine (7, 9). Steroid
glucuronidation activity increases along the length of the intestine
with the highest levels found in terminal ileum and colon
(35). Transport of E217G from mucosa to
serosa has been described in rat intestine (36) and
ATP-dependent transport of E217
G was markedly enhanced
in MRP3-transfected membrane vesicles (15).
Third, 1-naphthol and other phenolic compounds are glucuronidated by
the intestinal mucosa (2, 8, 12). Vectorial transport of
1-naphthol glucuronide from mucosal to serosal side occurs mainly in
the ileum and colon of rat intestine (20). In transport studies, 1-naphthol glucuronide has been shown to inhibit MRP3-mediated transport of E217G, indicating that this compound is
recognized by MRP3 (15). Fourth, flavonoids may be
glucuronidated to different extent in the intestine (7).
Transport of flavonoids from mucosa to serosa has been demonstrated in
vitro using intestinal Caco-2 cells (41, 42). Intestinal
loop experiments demonstrated carrier-mediated transport of riboflavin
in rat small and large intestine (43). Furthermore,
flavonoids may be potent inducers of MRP-ATPase activity (18), and flavonoid transport can be inhibited by the
MRP-inhibitor MK-571 (41, 42). In summary, this study is
the first demonstration that MRP3 protein is expressed in the rat
intestine. Unlike MRP2, which is predominantly expressed at the apical
domain of enterocytes from the proximal small intestine, MRP3 is highly
expressed at the basolateral membrane of enterocytes from ileum and
colon, where it may be involved in the ATP-dependent transport of
organic anions including bile salts, steroids, vitamins, and drugs from enterocytes into blood.
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ACKNOWLEDGEMENTS |
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The authors thank Dr. Dietrich Keppler for providing the EAG15 antibody and Dr. Jörg König for expert help with RT-PCR analysis.
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FOOTNOTES |
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This study was supported by the Forschungsförderungs-Programm (Projekt-Nr 199/1999) der Medizinischen Fakultät Heidelberg and by grants from the Deutsche Forschungsgemeinschaft through Sonderforschungsbereich 601.
Address for reprint requests and other correspondence: D. Rost, Dept. of Gastroenterology, Universitätsklinik Heidelberg. Bergheimer Strasse 58, D-69115 Heidelberg, Germany (E-mail: Daniel_Rost{at}med.uni-heidelberg.de).
The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
10.1152/ajpgi.00318.2001
Received 20 July 2001; accepted in final form 4 October 2001.
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REFERENCES |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
1.
Belinsky, MG,
Bain LJ,
Balsara BB,
Testa JR,
and
Kruh GD.
Characterization of MOAT-C and MOAT-D, new members of the MRP/cMOAT subfamily of transporter proteins.
J Natl Cancer Inst
90:
1735-1741,
1998
2.
Bock, KW,
and
Winne D.
Glucuronidation of 1-naphthol in the rat intestinal loop.
Biochem Pharmacol
24:
859-862,
1975[ISI][Medline].
3.
Borst, P,
Evers R,
Kool M,
and
Wijnholds J.
The multidrug resistance protein family.
Biochim Biophys Acta
1461:
347-357,
1999[ISI][Medline].
4.
Büchler, M,
König J,
Brom M,
Kartenbeck J,
Spring H,
Horie T,
and
Keppler D.
cDNA cloning of the hepatocyte canalicular isoform of the multidrug resistance protein, cMrp, reveals a novel conjugate export pump deficient in hyperbilirubinemic mutant rats.
J Biol Chem
271:
15091-15098,
1996
5.
Carey, MC,
and
Duane WC.
Enterohepatic circulation.
In: The Liver: Biology and Pathology, edited by Arias IM,
Boyer JL,
Fausto N,
Jakoby WB,
Schachter DA,
and Shafritz DA.. New York: Raven, 1994, p. 719-768.
6.
Catania, VA,
Luquita MG,
Sanchez Pozzi EJ,
and
Mottino AD.
Enhancement of intestinal UDP-glucuronosyltranferase activity in partially hepatectomized rats.
Biochim Biophys Acta
1380:
345-353,
1998[ISI][Medline].
7.
Cheng, Z,
Radominska-Pandya A,
and
Tephly TR.
Studies on the substrate specificity of human intestinal UDP-glucuronosyltransferases 1A8 and 1A10.
Drug Metab Dispos
27:
1165-1170,
1999
8.
Chowdhury, JR,
Novikoff PM,
Chowdhury NR,
and
Novikoff AB.
Distribution of UDPglucuronosyltransferase in rat tissue.
Proc Natl Acad Sci USA
82:
2990-2994,
1985[Abstract].
9.
Czernik, PJ,
Little JM,
Barone GW,
Raufman JP,
and
Radominska-Pandya A.
Glucuronidation of estrogens and retinoic acid and expression of UDP-glucuronosyltransferase 2B7 in human intestinal mucosa.
Drug Metab Dispos
28:
1210-1216,
2000
10.
De Vries, MH,
Hofman GA,
Koster AS,
and
Noordhoek J.
Systemic intestinal metabolism of 1-naphthol. A study in the isolated vascularly perfused rat small intestine.
Drug Metab Dispos
17:
573-578,
1989[Abstract].
11.
Gotoh, Y,
Suzuki H,
Kinoshita S,
Hirohashi T,
Kato Y,
and
Sugiyama Y.
Involvement of an organic anion transporter (canalicular multispecific organic anion transporter/multidrug resistance-associated protein 2) in gastrointestinal secretion of glutathione conjugates in rats.
J Pharmacol Exp Ther
292:
433-439,
2000
12.
Hartiala, K.
Metabolism of hormones, drugs and other substances by the gut.
Physiol Rev
53:
496-534,
1973
13.
Hirohashi, T,
Suzuki H,
Chu XY,
Tamai I,
Tsuji A,
and
Sugiyama Y.
Function and expression of multidrug resistance-associated protein family in human colon adenocarcinoma cells (Caco-2).
J Pharmacol Exp Ther
292:
265-270,
2000
14.
Hirohashi, T,
Suzuki H,
Ito K,
Ogawa K,
Kume K,
Shimizu T,
and
Sugiyama Y.
Hepatic expression of multidrug resistance-associated protein-like proteins maintained in eisai hyperbilirubinemic rats.
Mol Pharmacol
53:
1068-1075,
1998
15.
Hirohashi, T,
Suzuki H,
and
Sugiyama Y.
Characterization of the transport properties of cloned rat multidrug resistance-associated protein 3 (MRP3).
J Biol Chem
274:
15181-15185,
1999
16.
Hirohashi, T,
Suzuki H,
Takikawa H,
and
Sugiyama Y.
ATP-dependent transport of bile salts by rat multidrug resistance-associated protein 3 (Mrp3).
J Biol Chem
275:
2905-2910,
2000
17.
Hofmann, AF.
Intestinal absorption of bile acids and biliary constituents: the intestinal component of the enterohepatic circulation and the integrated system.
In: Physiology of the Gastrointestinal Tract, edited by Johnson LR.. New York: Raven, 1994, p. 1845-1865.
18.
Hooijberg, JH,
Broxterman HJ,
Scheffer GL,
Vrasdonk C,
Heijn M,
de Jong MC,
Scheper RJ,
Lankelma J,
and
Pinedo HM.
Potent interaction of flavopiridol with MRP1.
Br J Cancer
81:
269-276,
1999[ISI][Medline].
19.
Hopper, E,
Belinsky MG,
Zeng H,
Tosolini A,
Testa JR,
and
Kruh GD.
Analysis of the structure and expression pattern of MRP7 (ABCC10), a new member of the MRP subfamily.
Cancer Lett
162:
181-191,
2001[ISI][Medline].
20.
Inoue, H,
Yokota H,
Taniyama H,
Kuwahara A,
Ogawa H,
Kato S,
and
Yuasa A.
1-Naphthol -D-glucuronide formed intraluminally in rat small intestine mucosa and absorbed into the colon.
Life Sci
65:
1579-1588,
1999[ISI][Medline].
21.
Josting, D,
Winne D,
and
Bock KW.
Glucuronidation of paracetamol, morphine and 1-naphthol in the rat intestinal loop.
Biochem Pharmacol
25:
613-616,
1976[ISI][Medline].
22.
Keppler, D,
Kamisako T,
Leier I,
Cui Y,
Nies AT,
Tsujii H,
and
König J.
Localization, substrate specificity, and drug resistance conferred by conjugate export pumps of the MRP family.
Adv Enzyme Regul
40:
339-349,
2000[ISI][Medline].
23.
Kiuchi, Y,
Suzuki H,
Hirohashi T,
Tyson CA,
and
Sugiyama Y.
cDNA cloning and inducible expression of human multidrug resistance associated protein 3 (MRP3).
FEBS Lett
433:
149-152,
1998[ISI][Medline].
24.
König, J,
Nies AT,
Cui Y,
Leier I,
and
Keppler D.
Conjugate export pumps of the multidrug resistance protein (MRP) family: localization, substrate specificity, and MRP2-mediated drug resistance.
Biochim Biophys Acta
1461:
377-394,
1999[ISI][Medline].
25.
König, J,
Rost D,
Cui Y,
and
Keppler D.
Characterization of the human multidrug resistance protein isoform MRP3 localized to the basolateral hepatocyte membrane.
Hepatology
29:
1156-1163,
1999[ISI][Medline].
26.
Kool, M,
de Haas M,
Scheffer GL,
Scheper RJ,
van Eijk MJ,
Juijn JA,
Baas F,
and
Borst P.
Analysis of expression of cMOAT (MRP2), MRP3, MRP4, and MRP5, homologues of the multidrug resistance-associated protein gene (MRP1), in human cancer cell lines.
Cancer Res
57:
3537-3547,
1997[Abstract].
27.
Kool, M,
van der Linden M,
de Haas M,
Baas F,
and
Borst P.
Expression of human MRP6, a homologue of the multidrug resistance protein gene MRP1, in tissues and cancer cells.
Cancer Res
59:
175-182,
1999
28.
Kool, M,
van der Linden M,
de Haas M,
Scheffer GL,
de Vree JM,
Smith AJ,
Jansen G,
Peters GJ,
Ponne N,
Scheper RJ,
Elferink RP,
Baas F,
and
Borst P.
MRP3, an organic anion transporter able to transport anti-cancer drugs.
Proc Natl Acad Sci USA
96:
6914-6919,
1999
29.
Laemmli, UK.
Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
Nature
227:
680-685,
1970[ISI][Medline].
30.
Mottino, AD,
Hoffman T,
Jennes L,
Cao J,
and
Vore M.
Expression of multidrug resistance-associated protein 2 in small intestine from pregnant and postpartum rats.
Am J Physiol Gastrointest Liver Physiol
280:
G1261-G1273,
2001
31.
Mottino, AD,
Hoffman T,
Jennes L,
and
Vore M.
Expression and localization of multidrug resistant protein mrp2 in rat small intestine.
J Pharmacol Exp Ther
293:
717-723,
2000
32.
Ogawa, K,
Suzuki H,
Hirohashi T,
Ishikawa T,
Meier PJ,
Hirose K,
Akizawa T,
Yoshioka M,
and
Sugiyama Y.
Characterization of inducible nature of MRP3 in rat liver.
Am J Physiol Gastrointest Liver Physiol
278:
G438-G446,
2000
33.
Peng, KC,
Cluzeaud F,
Bens M,
Van Huyen JP,
Wioland MA,
Lacave R,
and
Vandewalle A.
Tissue and cell distribution of the multidrug resistance-associated protein (MRP) in mouse intestine and kidney.
J Histochem Cytochem
47:
757-768,
1999
34.
Pinkus, LM,
Ketley JN,
and
Jakoby WB.
The glutathione S-transferases as a possible detoxification system of rat intestinal epithelium.
Biochem Pharmacol
26:
2359-2363,
1977[ISI][Medline].
35.
Radominska-Pandya, A,
Little JM,
Pandya JT,
Tephly TR,
King CD,
Barone GW,
and
Raufman JP.
UDP-glucuronosyltransferases in human intestinal mucosa.
Biochim Biophys Acta
1394:
199-208,
1998[ISI][Medline].
36.
Schwenk, M,
Schiemenz C,
del Pino VL,
and
Remmer H.
First pass biotransformation of ethinylestradiol in rat small intestine in situ.
Naunyn Schmiedebergs Arch Pharmacol
321:
223-225,
1982[ISI][Medline].
37.
Soroka, CJ,
Lee JM,
Azzaroli F,
and
Boyer JL.
Cellular localization and up-regulation of multidrug resistance-associated protein 3 in hepatocytes and cholangiocytes during obstructive cholestasis in rat liver.
Hepatology
33:
783-791,
2001[ISI][Medline].
38.
Suzuki, H,
and
Sugiyama Y.
Role of metabolic enzymes and efflux transporters in the absorption of drugs from the small intestine.
Eur J Pharm Sci
12:
3-12,
2000[ISI][Medline].
39.
Tammur, J,
Prades C,
Arnould I,
Rzhetsky A,
Hutchinson A,
Adachi M,
Schuetz JD,
Swoboda KJ,
Ptacek LJ,
Rosier M,
Dean M,
and
Allikmets R.
Two new genes from the human ATP-binding cassette transporter superfamily, ABCC11 and ABCC12, tandemly duplicated on chromosome 16q12.
Gene
273:
89-96,
2001[ISI][Medline].
40.
Van Aubel, RA,
Hartog A,
Bindels RJ,
Van Os CH,
and
Russel FG.
Expression and immunolocalization of multidrug resistance protein 2 in rabbit small intestine.
Eur J Pharmacol
400:
195-198,
2000[ISI][Medline].
41.
Walgren, RA,
Karnaky KJ, Jr,
Lindenmayer GE,
and
Walle T.
Efflux of dietary flavonoid quercetin 4'--glucoside across human intestinal Caco-2 cell monolayers by apical multidrug resistance-associated protein-2.
J Pharmacol Exp Ther
294:
830-836,
2000
42.
Walle, UK,
French KL,
Walgren RA,
and
Walle T.
Transport of genistein-7-glucoside by human intestinal CACO-2 cells: potential role for MRP2.
Res Commun Mol Pathol Pharmacol
103:
45-56,
1999[ISI][Medline].
43.
Yuasa, H,
Hirobe M,
Tomei S,
and
Watanabe J.
Carrier-mediated transport of riboflavin in the rat colon.
Biopharm Drug Dispos
21:
77-82,
2000[ISI][Medline].
44.
Zeng, H,
Liu G,
Rea PA,
and
Kruh GD.
Transport of amphipathic anions by human multidrug resistance protein 3.
Cancer Res
60:
4779-4784,
2000