First evidence of the P-glycoprotein gene expression and multixenobiotic resistance modulation in earthworm

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Multixenobiotic resistance (MXR) is an important mechanism of cellular efflux mediated by ATP binding cassette (ABC) transporters that bind and actively remove toxic substrates from the cell. This study was the first to identify ABC transporter P-glycoprotein (P-gp/ABCB1) as a representative of the MXR phenotype in earthworm (Eisenia fetida). The identified partial cDNA sequence of ABCB1 overlapped with ABCB1 homologues of other organisms from 58.5 % to 72.5 %. We also studied the effect of five modulators (verapamil, cyclosporine A, MK571, probenecid, and orthovanadate) on the earthworm’s MXR activity by measuring the accumulation of model substrates rhodamine B and rhodamine 123 in whole body tissue of the adult earthworm. MK571, orthovanadate, and verapamil significantly inhibited MXR activity, and rhodamine 123 turned out to better reflect MXR activity in that species than rhodamine B. Our results show that E. fetida can serve well as a test organism for environmental pollutants that inhibit MXR activity.

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  • 1. Luo Y Zang Y Zhong Y Kong Z. Toxicological study of two novel pesticides on earthworm Eisenia foetida. Chemosphere 1999;39:2347-56. doi: 10.1016/S0045-6535(99)00142-3

  • 2. Markman S Guschina IA Barnsley S Buchanan KL Pascoe D Müller CT. Endocrine disrupting chemicals accumulate in earthworms exposed to sewage effluent. Chemosphere 2007;70:119-25. doi: 10.1016/j.chemosphere.2007.06.045

  • 3. Garcia M. Römbke J de Brito MT Scheffczyk A. Effects of three pesticides on the avoidance behavior of earthworms in laboratory tests performed under temperate and tropical conditions. Environ Pollut 2008;153:450-6. doi:10.1016/j. envpol.2007.08.007

  • 4. Sinha R. Earthworms: the miracle of nature (Charles Darwin’s ‘unheralded soldiers of mankind & farmer’s friends’). The Environmentalist 2009;29:339-40. doi: 10.1007/s10669-009-9242-4

  • 5. Organisation for Economic Cooperation and Development (OECD). OECD Guidelines for Testing of Chemicals.Section 2. Effects on Biotic Systems. Test No. 207: Earthworm Acute Toxicity Tests (Filter Paper Test and Artificial Soil Test). Paris: OECD; 1984.

  • 6. Goldstone JV Hamdoun A Cole BJ Howard-Ashby M Nebert DW Scally M Dean M Epel D Hahn ME Stegeman JJ. The chemical defensome: environmental sensing and response genes in the Strongylocentrotus purpuratus genome.Dev Biol 2006;300:366-84. doi: 1016/j.ydbio.2006.08.066

  • 7. Epel D Luckenbach T Stevenson CN MacManus-Spencer LA Hamdoun A Smital T. Efflux transporters: newly appreciated roles in protection against pollutants. Environ Sci Technol 2008;42:3914-20. PMID: 18589945

  • 8. Sauerborn Klobučar R Žaja R Franjević D Brozović A Smital T. Presence of ecotoxicologically relevant P-gp and MRP transcripts and proteins in cyprinid fish. Arh Hig Rada Toksikol 2010;61:175-82. doi: 10.2478/10004-1254-61-2010-2008

  • 9. Bošnjak I Uhlinger KR Heim W Smital T Franekić Čolić J Coale K Epel D Hamdoun A. Multidrug efflux transporters limit accumulation of inorganic but not organic mercury in sea urchin embryos. Environ Sci Technol 2009;43:8374-80. doi: 10.1021/es901677r

  • 10. Hackenberger BK Velki M Stepic S Hackenberger DK.First evidence for the presence of efflux pump in the earthworm Eisenia andrei. Ecotoxicol Environ Saf 2012;75:40-5. doi: 10.1016/j.ecoenv.2011.06.024

  • 11. Leslie EM Deeley RG Cole SPC. Multidrug resistance proteins: role of P-glycoprotein MRP1 MRP2 and BCRP (ABCG2) in tissue defense. Toxicol Appl Pharmacol 2005;204:216-37. PMID: 15845415

  • 12. Szakács G Váradi A Özvegy-Laczka C Sarkadi B. The role of ABC transporters in drug absorption distribution metabolism excretion and toxicity (ADME-Tox). Drug Discov Today 2008;13:379-93. doi: 10.1016/j. drudis.2007.12.010

  • 13. Lespine A Ménez C Bourguinat C Prichard RK.P-glycoproteins and other multidrug resistance transporters Bošnjak I et al. P-GP/ABCB1 AND MXR ACTIVITY IN E. FETIDA Arh Hig Rada Toksikol 2014;65:67-75 in the pharmacology of anthelmintics: Prospects for reversing transport-dependent anthelmintic resistance. Int J Parasitol Drugs Drug Resist 2012;2:58-75. doi: 10.1016/j. ijpddr.2011.10.001

  • 14. Kurelec B Krča S Pivčević B Ugarković D Bachmann M Imsiecke G Müller WEG. Expression of P-glycoprotein gene in marine sponges. Identification and characterization of the 125 kDa drug-binding glycoprotein. Carcinogenesis 1992;13:69-76. doi: 10.1093/carcin/13.1.69

  • 15. Minier C Akcha F Galgani F. P-glycoprotein expression in Crassostrea gigas and Mytilus edulis in polluted seawater.Comp Biochem Physiol B Biochem Mol Biol 1993;106:1029-36. PMID: 7905372

  • 16. Smital T Sauerborn R Pivčević B Krča S Kurelec B.Interspecies differences in P-glycoprotein mediated activity of multixenobiotic resistance mechanism in several marine and freshwater invertebrates. Comp Biochem Physiol C Pharmacol Toxicol Endocrinol 2000;126:175-86. PMID: 11050689

  • 17. Bard SM Woodin BR Stegeman JJ. Expression of P-glycoprotein and cytochrome p450 1A in intertidal fish (Anoplarchus purpurescens) exposed to environmental contaminants. Aquat Toxicol 2002;60:17-32. doi: 10.1016/ S0166-445X(01)00272-7

  • 18. Smital T Sauerborn R Hackenberger BK. Inducibility of the P-glycoprotein transport activity in the marine mussel Mytilus galloprovincialis and the freshwater mussel Dreissena polymorpha. Aquat Toxicol 2003;65:443-65. doi: 10.1016/S0166-445X(03)00175-9

  • 19. Hamdoun AM Cherr GN Roepke TA Epel D. Activation of multidrug efflux transporter activity at fertilization in sea urchin embryos (Strongylocentrotus purpuratus). Dev Biol 2004;276:452-62. doi: 10.1016/j.ydbio.2004.09.013

  • 20. Sinha RK Chauhan K Valani D Chandran V Soni BK Patel V. Earthworms: Charles Darwin’s “unheralded soldiers of mankind”: Protective & productive for man & environment.J Environ Prot 2010;1:251-60. doi: 10.4236/jep.2010.13030

  • 21. Sambrook J Fritsch EF Maniatis T. Molecular Cloning: a Laboratory Manual. Cold Spring Harbor (NY): Cold Spring Harbor Laboratory; 1989.

  • 22. Vogelstein B Gillespie D. Preparative and analytical purification of DNA from agarose. Proc Natl Acad Sci USA 1979;76:615-9. PMCID: PMC382999

  • 23. Knoche K Kephart D. Cloning blunt-end Pfu DNA Polymerase-generated PCR fragments into pGEM®-T Vector Systems. Promega Notes 1999;71:10-3.

  • 24. Tamura K Peterson D Peterson N Stecher G Nei M Kumar S. MEGA5: molecular evolutionary genetics analysis using maximum likelihood evolutionary distance and maximum parsimony methods. Mol Biol Evol 2011;28:2731-9. doi: 10.1093/molbev/msr121

  • 25. Hillis DM Bull JJ. An empirical test of bootstrapping as a method for assessing confidence in phylogenetic analysis.Syst Biol 1993;42:182-92. doi: 10.1093/sysbio/42.2.182

  • 26. Le SQ Gascuel O. An improved general amino-acid replacement matrix. Mol Biol Evol 2008;25:1307-20. doi: 10.1093/molbev/msn067

  • 27. Abascal F Zardoya R Posada D. ProtTest: Selection of bestfit models of protein evolution. Bioinformatics 2005;21:2104-5. doi: 10.1093/bioinformatics/bti263

  • 28. Guindon S Dufayard JF Lefort V Anisimova M Hordijk W Gascuel O. New algorithms and methods to estimate maximum-likelihood phylogenies: assessing the performance of PhyML 3.0. Syst Biol 2010;59:307-21. doi: 10.1093/ sysbio/syq010

  • 29. Hordijk W Gascuel O. Improving the efficiency of SPR moves in phylogenetic tree search methods based on maximum likelihood. Bioinformatics 2005;21:4338-47. doi: 10.1093/bioinformatics/bti713

  • 30. Anisimova M Gascuel O. Approximate likelihood-ratio test for branches: a fast accurate and powerful alternative. Syst Biol 2006;55:539-52. doi: 10.1080/10635150600755453

  • 31. Dean M Rzhetsky A Allikmets R. The human ATP-binding cassette (ABC) transporter superfamily. Genome Res 2001;11:1156-66. doi: 10.1101/gr.184901

  • 32. Fischer E Beuschlein F Bidlingmaier M Reincke M.Commentary on the endocrine society practice guidelines: Consequences of adjustment of antihypertensive medication in screening of primary aldosteronism. Rev Endocr Metab Disord 2011;12:43-8. doi: 10.1007/s11154-011-9163-7

  • 33. Gekeler V Ise W Sanders KH Ulrich WR Beck J. The leukotriene LTD4 receptor antagonist Mk571 specifically modulates MRP associated multidrug resistance. Biochem Biophys Res Commun 1995;208:345-52. PMID: 7887949

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