Abstract
Rumen fermentation of plant-based forage in bovines is the major site for generation and absorption of short-chain fatty acids. Consequentially, the rumen is also the site for initial exposure to toxins released from diet. Accordingly, we have investigated the expression of bovine ABC transporters in the rumen associated with cytoprotection against xenobiotic exposure, namely MDR1 (ABCB1), MRP2 (ABCC2) and BCRP (ABCG2). Bovine rumen samples from the ventral sac were obtained post-mortem from a commercial slaughterhouse after humane killing. Rumen papilla samples were then prepared for total RNA isolation for RT-PCR, SDS-PAGE/Western blotting and immunohistochemistry. PCR products of the predicted size were observed for both MDR1 and BCRP, but not for MRP2 using bovine-specific primers. β-actin was used as a control transcript. Western blot analysis using C219 primary monoclonal antibody revealed MDR1 protein expression in bovine rumen (Mapp, of ~170–180 kD). Immunolocalisation of MDR1 using UIC2 monoclonal antibody within cryosections of bovine rumen showed extensive membrane staining in the cells of the stratum granulosum, stratum spinosum and stratum basale. MDR1 expression was absent from outer stratum corneum. Protein expression and immunolocalisation were also confirmed for BCRP, with prevalent staining in the stratum basale, becoming weaker in the stratum spinosum and stratum granulosum.
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Altschul SF et al (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402
Aschenbach JR, Penner GB, Stumpff F, Gabel G (2011) Ruminant nutrition symposium: role of fermentation acid absorption in the regulation of ruminal pH. J Anim Sci 89:1092–1107
Ballent M, Lifschitz A, Virkel G, Sallovitz J, Lanusse C (2007) Involvement of P-glycoprotein on ivermectin kinetic behaviour in sheep: itraconazole-mediated changes on gastrointestinal disposition. J Veterinary Pharmacol Ther 30:242–248
Brayden DJ, Griffin J (2008) Avermectin transepithelial transport in MDR1- and MRP-transfected canine kidney monolayers. Veterinary Res Commun 32:93–106
Bunting KD (2002) ABC transporters as phenotypic markers and functional regulators of stem cells. Stem Cells 20:11–20
Chan LMS, Lowes S, Hirst BH (2004) The ABCs of drug transport in intestine and liver: efflux proteins limiting drug absorption and bioavailability. Eur J Pharm Sci 21:25–51
Cordoncardo C et al (1989) Multidrug-resistance gene (P-Glycoprotein) is expressed by endothelial-cells at blood–brain barrier sites. Proc Natl Acad Sci USA 86:695–698
Dietrich CG, Geier A, Elferink RPJO (2003) ABC of oral bioavailability: transporters as gatekeepers in the gut. Gut 52:1788–1795
Eisenblatter T, Huwel S, Galla HJ (2003) Characterisation of the brain multidrug resistance protein (BMDP/ABCG2/BCRP) expressed at the blood–brain barrier. Brain Res 971:221–231
Gabel G, Sehested J (1997) SCFA transport in the forestomach of ruminants. Comp Biochem Physiol A Physiol 118:367–374
Goncalves P, Gregorio I, Martel F (2011) The short-chain fatty acid butyrate is a substrate of breast cancer resistance protein. Am J Physiol Cell Physiol 301:C984–C994
Graham C, Simmons NL (2005) Functional organization of the bovine rumen epithelium. Am J Physiol Regul Integr Comp Physiol 288:R173–R181
Graham C, Gatherar I, Haslam I, Glanville M, Simmons NL (2007) Expression and localization of monocarboxylate transporters and sodium/proton exchangers in bovine rumen epithelium. Am J Physiol Regul Integr Comp Physiol 292:R997–R1007
Greer DA, Ivey S (2007) Distinct N-glycan glycosylation of P-glycoprotein isolated from the human uterine sarcoma cell line MES-SA/Dx5. Biochim Biophys Acta 1770:1275–1282
Ito K et al (2008) P-Glycoprotein (Abcb1) is involved in absorptive drug transport in skin. J Control Release 131:198–204
Lindner S, Halwachs S, Wassermann L, Honscha W (2013) Expression and subcellular localization of efflux transporter ABCG2/BCRP in important tissue barriers of lactating dairy cows, sheep and goats. J Vet Pharmacol Ther 36(6):562–570
Maliepaard M et al (2001) Subcellular localization and distribution of the breast cancer resistance protein transporter in normal human tissues. Cancer Res 61:3458–3464
Mechetner EB, Roninson IB (1992) Efficient inhibition of P-glycoprotein-mediated multidrug resistance with a monoclonal-antibody. Proc Natl Acad Sci USA 89:5824–5828
Murakami T, Takano M (2008) Intestinal efflux transporters and drug absorption. Expert Opin Drug Met 4:923–939
Prichard RK, Steel JW, Lacey E, Hennessy DR (1985) Pharmacokinetics of ivermectin in sheep following intravenous, intra-abomasal or intraruminal administration. J Vet Pharmacol Ther 8:88–94
Prouillac C, Videmann B, Mazallon M, Lecoeur S (2009) Induction of cells differentiation and ABC transporters expression by a myco-estrogen, zearalenone, in human choriocarcinoma cell line (BeWo). Toxicology 263:100–107
Rabbani I, Siegling-Vlitakis C, Noci B, Martens H (2011) Evidence for NHE3-mediated Na transport in sheep and bovine forestomach. Am J Physiol Regul Integr Comp Physiol 301:R313–R319
Rea PA, Li ZS, Lu YP, Drozdowicz YM, Martinoia E (1998) From vacuolar Gs-X pumps to multispecific ABC Transporters. Annu Rev Plant Physiol Plant Mol Biol 49:727–760
Ritter JK (2007) Intestinal UGTs as potential modifiers of pharmacokinetics and biological responses to drugs and xenobiotics. Expert Opin Drug Metab 3:93–107
Schinkel AH, Kemp S, Dolle M, Rudenko G, Wagenaar E (1993) N-glycosylation and deletion mutants of the human MDR1 P-glycoprotein. J Biol Chem 268:7474–7481
Schinkel AH et al (1994) Disruption of the mouse mdr1a P-glycoprotein gene leads to a deficiency in the blood–brain barrier and to increased sensitivity to drugs. Cell 77:491–502
Shugarts S, Benet LZ (2009) The role of transporters in the pharmacokinetics of orally administered drugs. Pharm Res 26:2039–2054
Skazik C et al (2011) P-glycoprotein (ABCB1) expression in human skin is mainly restricted to dermal components. Exp Dermatol 20:450–452
Thelen K, Dressman JB (2009) Cytochrome P450-mediated metabolism in the human gut wall. J Pharm Pharmacol 61:541–558
Wassermann L, Halwachs S, Lindner S, Honscha KU, Honscha W (2013) Determination of functional ABCG2 activity and assessment of drug-ABCG2 interactions in dairy animals using a novel MDCKII in vitro model. J Pharm Sci 102:772–784
Zhang YC, Benet LZ (2001) The gut as a barrier to drug absorption—combined role of cytochrome P450 3A and P-glycoprotein. Clin Pharmacokinet 40:159–168
Zhang P, Tian X, Chandra P, Brouwer KL (2005) Role of glycosylation in trafficking of Mrp2 in sandwich-cultured rat hepatocytes. Mol Pharmacol 67:1334–1341
Zhou Y, Gottesman MM, Pastan I (1999) The extracellular loop between TM5 and TM6 of P-glycoprotein is required for reactivity with monoclonal antibody UIC2. Arch Biochem Biophys 367:74–80
Acknowledgments
Iain Haslam was supported by a studentship from the BBSRC (UK). This work was part funded by Pfizer Global Research Sandwich UK.
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Communicated by H.V. Carey.
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Haslam, I.S., Simmons, N.L. Expression of the ABC transport proteins MDR1 (ABCB1) and BCRP (ABCG2) in bovine rumen. J Comp Physiol B 184, 673–681 (2014). https://doi.org/10.1007/s00360-014-0804-5
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DOI: https://doi.org/10.1007/s00360-014-0804-5