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Excretion of fluorescent substrates of mammalian multidrug resistance-associated protein (MRP) in the Schistosoma mansoni excretory system

Published online by Cambridge University Press:  19 January 2004

H. SATO
Affiliation:
Division of Biochemistry and Molecular Biology, Institute of Biomedical and Life Sciences, West Medical Building, University of Glasgow, Glasgow G12 8QQ, Scotland present address: Department of Parasitology, Hirosaki University School of Medicine, Hirosaki 036-8562, Japan. Fax: +81 172 39 5045. E-mail: [email protected]
J. R. KUSEL
Affiliation:
Division of Biochemistry and Molecular Biology, Institute of Biomedical and Life Sciences, West Medical Building, University of Glasgow, Glasgow G12 8QQ, Scotland
J. THORNHILL
Affiliation:
Division of Biochemistry and Molecular Biology, Institute of Biomedical and Life Sciences, West Medical Building, University of Glasgow, Glasgow G12 8QQ, Scotland

Abstract

The protonephridium of platyhelminths including Schistosoma mansoni plays a pivotal role in their survival by excretion of metabolic wastes as well as xenobiotics, and can be revealed in the living adult parasite by certain fluorescent compounds which are concentrated in excretory tubules and collecting ducts. To determine the presence of the multidrug resistance-associated protein (MRP) as a possible transporter in protonephridial epithelium, adult schistosomes were exposed to a fluorescent Ca2+ indicator, fluo-3 acetyloxymethyl ester, which is a potential substrate of mammalian MRP. Specific fluorescence related to fluo-3/Ca2+ chelate delineated the whole length of the protonephridial system. Simultaneously, a fluorescent substance was accumulated in the posterior part of collecting ducts and the excretory bladder. Similarly, when other fluorogenic substrates for mammalian MRP such as monoclorobimane, fluorescein diacetate, and 5(6)-carboxyfluorescein diacetate were applied to adult schistosomes, these fluorescent markers were observed in the excretory tubules through to the excretory bladder. The excretory system of mechanically-transformed schistosomula was not labelled with any of these 4 fluorescent markers. These findings suggest that the protonephridial epithelium of adult schistosomes, but not schistosomula, might express the homologue of the mammalian MRP transporting organic anionic conjugates with glutathione, glucuronate or sulphate as well as unconjugated amphiphilic organic anions.

Type
Research Article
Copyright
2004 Cambridge University Press

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References

REFERENCES

ABRAHAMSE, S. L. & RECHKEMMER, G. (2001). Identification of an organic anion transport system in the human colon carcinoma cell line HT29 clone 19A. Pflügers Archiv: European Journal of Physiology 441, 529537.CrossRefGoogle Scholar
AMBUDKAR, S. V., DEY, S., HRYCYNA, C. A., RAMACHANDRA, M., PASTAN, I. & GOTTESMAN, M. M. (1999). Biochemical, cellular, and pharmacological aspects of the multidrug transporter. Annual Review of Pharmacology and Toxicology 39, 361398.CrossRefGoogle Scholar
BORST, P., EVERS, R., KOOL, M. & WIJNHOLDS, J. (1999). The multidrug resistance protein family. Biochimica et Biophysica Acta 1461, 347357.CrossRefGoogle Scholar
BOSCH, I. B., WANG, Z.-X., TAO, L.-F. & SHOEMAKER, C. B. (1994). Two Schistosoma mansoni cDNAs encoding ATP-binding cassette (ABC) family proteins. Molecular and Biochemical Parasitology 65, 351356.CrossRefGoogle Scholar
BROEKS, A., GERRARD, B., ALLIKMETS, R., DEAN, M. & PLASTERK, R. H. A. (1996). Homologues of the human multidrug resistance genes MRP and MDR contribute to heavy metal resistance in the soil nematode Caenorhabditis elegans. EMBO Journal 15, 61326143.Google Scholar
CANTZ, T., NIES, A. T., BROM, M., HOFMANN, A. F. & KEPPLER, D. (2000). MRP2, a human conjugate export pump, is present and transports fluo-3 into apical vacuoles of Hep G2 cells. American Journal of Physiology: Gastrointestinal and Liver Physiology 278, G522G531.Google Scholar
COURTOIS, A., PAYEN, L., LAGADIC, D., GUILLOUZO, A. & FARDEL, O. (1999). Evidence for a multidrug resistance-associated protein 1 (MRP1)-related transport system in cultured rat liver biliary epithelial cells. Life Sciences 64, 763774.CrossRefGoogle Scholar
ENDICOTT, J. A. & LING, V. (1989). The biochemistry of P-glycoprotein-mediated multidrug resistance. Annual Review of Biochemistry 58, 137171.CrossRefGoogle Scholar
FALLON, P. G., TAO, L.-F., ISMAIL, M. M. & BENNETT, J. L. (1996). Schistosome resistance to praziquantel: fact or artifact? Parasitology Today 12, 316320.Google Scholar
FELLER, N., BROXTERMAN, H. J., WÄHRER, D. C. R. & PINEDO, H. M. (1995). ATP-dependent efflux of calcein by the multidrug resistance protein (MRP): no inhibition by intracellular glutathione depletion. FEBS Letters 368, 385388.CrossRefGoogle Scholar
GENAZZANI, A. A. & GALIONE, A. (1996). Nicotinic acid-adenine dinucleotide phosphate mobilizes Ca2+ from a thapsigargin-insensitive pool. The Biochemical Journal 315, 721725.CrossRefGoogle Scholar
HAUGLAND, R. P. (1996). Handbook of Fluorescent Probes and Research Chemicals, 6th Edn. Molecular Probes Inc., Eugene, Oregon.
HIGGINS, C. F. (1992). ABC transporters: from microorganisms to man. Annual Review of Cell Biology 8, 67113.CrossRefGoogle Scholar
HIPFNER, D. R., DEELEY, R. G. & COLE, S. P. C. (1999). Structural, mechanistic and clinical aspects of MRP1. Biochimica et Biophysica Acta 1461, 359376.CrossRefGoogle Scholar
HOLLÓ, Z., HOMOLYA, L., HEGEDÜS, T. & SARKADI, B. (1996). Transport properties of the multidrug resistance-associated protein (MRP) in human tumour cells. FEBS Letters 383, 99104.CrossRefGoogle Scholar
HOMOLYA, L., HOLLÓ, Z., GERMANN, U. A., PASTAN, I., GOTTESMAN, M. M. & SARKADI, B. (1993). Fluorescent cellular indicators are extruded by the multidrug resistance protein. Journal of Biological Chemistry 268, 2149321496.Google Scholar
HOPPER, E., BELINSKY, M. G., ZENG, H., TOSOLINI, A., TESTA, J. R. & KRUH, G. D. (2001). Analysis of the structure and expression pattern of MRP7 (ABCC10), a new member of the MRP subfamily. Cancer Letters 162, 181191.CrossRefGoogle Scholar
HUANG, Y.-J. & PRICHARD, R. K. (1999). Identification and stage-specific expression of two putative P-glycoprotein coding genes in Onchocerca volvulus. Molecular and Biochemical Parasitology 102, 273281.CrossRefGoogle Scholar
KAO, J. P. Y., HAROOTUNIAN, A. T. & TSIEN, R. Y. (1989). Photochemically generated cytosolic calcium pulses and their detection by Fluo-3. Journal of Biological Chemistry 264, 81798184.Google Scholar
KEPPLER, D. & KÖNIG, J. (1997). Expression and localization of the conjugate export pump encoded by the MRP2 (cMRP/cMOAT) gene in liver. FASEB Journal 11, 509516.CrossRefGoogle Scholar
KEPPLER, D., CUI, Y., KÖNING, J., LEIER, I. & NIES, A. (1999). Export pumps for anionic conjugates encoded by MRP genes. Advances in Enzyme Regulation 39, 237246.CrossRefGoogle Scholar
KLEIN, I., SARKADI, B. & VÁRADI, A. (1999). An inventory of the human ABC proteins. Biochimica et Biophysica Acta 1461, 237262.CrossRefGoogle Scholar
KÖNIG, J., NIES, A. T., CUI, Y., LEIER, I. & KEPPLER, D. (1999). Conjugate export pumps of the multidrug resistance protein (MRP) family: localization, substrate specificity, and MRP2-mediated drug resistance. Biochimica et Biophysica Acta 1461, 377394.CrossRefGoogle Scholar
KUSEL, J. R. & HAGAN, P. (1999). Praziquantel: its use, cost and possible development of resistance. Parasitology Today 15, 352354.CrossRefGoogle Scholar
LAUPEZE, B., AMIOT, L., COURTOIS, A., VERNHET, L., DRENOU, B., FAUCHET, R. & FARDEL, O. (1999). Use of the anionic dye carboxy-2′,7′-dichlorofluorescein for sensitive flow cytometric detection of multidrug resistance-associated protein activity. International Journal of Oncology 15, 571576.CrossRefGoogle Scholar
LOHOFF, M., PRECHTL, S., SOMMER, F., ROELLINGHOFF, M., SCHMITT, E., GRADEHANDT, G., ROHWER, P., STRIDE, B. D., COLE, S. P. C. & DEELEY, R. G. (1998). A multidrug-resistance protein (MRP)-like transmembrane pump is highly expressed by resting murine T helper(Th)2, but not Th1 cells, and is induced to equal expression levels in Th1 and Th2 cells after antigenic stimulation in vivo. Journal of Clinical Investigation 101, 703710.CrossRefGoogle Scholar
MINTA, A., KAO, J. P. Y. & TSIEN, R. Y. (1989). Fluorescent indicators for cytosolic calcium based on rhodamine and fluorescein chromophores. Journal of Biological Chemistry 264, 81718178.Google Scholar
NIEMANN, A., BALTES, J. & ELSÄSSER, H.-P. (2001). Fluorescence properties and staining behaviour of monodansylpentane, a structural homologue of the lysosomotropic agent monodansylcadaverine. Journal of Histochemistry and Cytochemistry 49, 177185.CrossRefGoogle Scholar
NIES, A. T., CANTZ, T., BROM, M., LEIER, I. & KEPPLER, D. (1998). Expression of the apical conjugate export pump, Mrp2, in the polarized hepatoma cell line, WIF-B. Hepatology 28, 13321340.CrossRefGoogle Scholar
OUDE ELFERINK, R. P. J., BAKKER, C. T. M., ROELOFSEN, H., MIDDELKOOP, E., OTTENHOFF, R., HEIJN, M. & JANSEN, P. L. M. (1993). Accumulation of organic anion in intracellular vesicles of cultured rat hepatocytes is mediated by the canalicular multispecific organic anion transporter. Hepatology 17, 434444.CrossRefGoogle Scholar
PAPADOPOULOU, B., ROY, G., DEY, S., ROSEN, B. P. & OUELLETTE, M. (1994). Contribution of the Leishmania P-glycoprotein-related gene ltpgpA to oxyanion resistance. Journal of Biological Chemistry 269, 1198011986.Google Scholar
PEARCE, E. J. & MacDONALD, A. S. (2002). The immunobiology of schistosomiasis. Nature Reviews 2, 499511.CrossRefGoogle Scholar
REED, M. B., PANACCIO, M., STRUGNELL, R. A. & SPITHILL, T. W. (1998). Developmental expression of a Fasciola hepatica sequence homologous to ABC transporters. International Journal for Parasitology 28, 13751381.CrossRefGoogle Scholar
RIBEIRO, F., COELHO, P. M. Z., VIEIRA, L. Q., WATSON, D. G. & KUSEL, J. R. (1998). The effect of praziquantel treatment on glutathione concentration in Schistosoma mansoni. Parasitology 116, 229236.CrossRefGoogle Scholar
ROELOFSEN, H., BAKKER, C. T. M., SCHOEMAKER, B., HEIJN, M., JANSEN, P. L. M. & OUDE ELFERINK, R. P. J. (1995). Redistribution of canalicular organic anion transport activity in isolated and cultured rat hepatocytes. Hepatology 21, 16491657.Google Scholar
ROELOFSEN, H., SOROKA, C. J., KEPPLER, D. & BOYER, J. L. (1998). Cyclic AMP stimulates sorting of the canalicular organic anion transporter (Mrp2/cMoat) to the apical domain in hepatocyte couplets. Journal of Cell Science 111, 11371145.Google Scholar
SANGSTER, N. C., BANNAN, S. C., WEISS, A. S., NULF, S. C., KLEIN, R. D. & GEARY, T. G. (1999). Haemonchus contortus: sequence heterogeneity of internucleotide binding domains from P-glycoproteins and an association with avermectin/milbemycin resistance. Experimental Parasitology 91, 250257.CrossRefGoogle Scholar
SATO, H., KUSEL, J. R. & THORNHILL, J. (2002). Functional visualization of the excretory system of adult Schistosoma mansoni by the fluorescent marker resorufin. Parasitology 125, 527535.CrossRefGoogle Scholar
SCHAUB, T. P., KARTENBECK, J., KÖNIG, J., SPRING, H., DÖRSAM, J., STAEHLER, G., STÖRKEL, S., THON, W. F. & KEPPLER, D. (1999). Expression of the MRP2 gene-encoded conjugate export pump in human kidney proximal tubules and in renal-cell carcinoma. Journal of the American Society of Nephrology 10, 11591169.Google Scholar
SCHAUB, T. P., KARTENBECK, J., KÖNIG, J., VOGEL, O., WITZGALL, R., KRIZ, W. & KEPPLER, D. (1997). Expression of the conjugate export pump encoded by the mrp2 gene in the apical membrane of kidney proximal tubules. Journal of the American Society of Nephrology 8, 12131221.Google Scholar
SMITHERS, S. R. & TERRY, R. J. (1965). The infection of laboratory hosts with cercariae of Schistosoma mansoni and the recovery of adult worms. Parasitology 55, 695700.CrossRefGoogle Scholar
SZCZYPKA, M. S., WEMMIE, J. A., MOYE-ROWLEY, W. S. & THIELE, D. J. (1994). A yeast metal resistance protein similar to human cystic fibrosis transmembrane conductance regulator (CFTR) and multidrug resistance-associated protein. Journal of Biological Chemistry 269, 2285322857.Google Scholar
TAN, H. H. C., THORNHILL, J. A., AL-ADHAMI, B. H., AKHKHA, A. A. & KUSEL, J. R. (2003). A study of the effect of surface damage on the uptake of Texas Red-BSA by schistosomula of Schistosoma mansoni. Parasitology 126, 235240.CrossRefGoogle Scholar
TREIMAN, M., CASPERSEN, C. & CHRISTENSEN, S. B. (1998). A tool coming of age: thapsigargin as an inhibitor of sarcoendeplasmic reticulum Ca2+-ATPases. Trends in Pharmacological Sciences 19, 131135.CrossRefGoogle Scholar
ULLMAN, B. (1995). Multidrug resistance and P-glycoproteins in parasitic protozoa. Journal of Bioenergetics and Biomembranes 27, 7784.CrossRefGoogle Scholar
UPCROFT, P. (1994). Multiple drug resistance in the pathogenic protozoa. Acta Tropica 56, 195212.CrossRefGoogle Scholar
VAN DER KOLK, D. M., DE VRIES, E. G. E., KONING, J. A., VAN DEN BERG, E., MÜLLER, M. & VELLENGA, E. (1998). Activity and expression of the multidrug resistance proteins MRP1 and MRP2 in acute myeloid leukemia cells, tumor cell lines, and normal hematopoietic CD34+ peripheral blood cells. Clinical Cancer Research 4, 17271736.Google Scholar
WILSON, R. A. & WEBSTER, L. A. (1974). Protonephridia. Biological Reviews 49, 127160.CrossRefGoogle Scholar
WIPPERSTEG, V., RIBEIRO, F., LIEDTKE, S., KUSEL, J. R. & GREVELDING, C. G. (2003). The uptake of Texas Red BSA in the excretory system of schistosomes and its colocalisation with ER60 promoter induced GFP in transiently transformed adult males. International Journal for Parasitology 33, 11391143.CrossRefGoogle Scholar
XU, M., MOLENTO, M., BLACKHALL, W., RIBEIRO, P., BEECH, R. & PRICHARD, R. (1998). Ivermectin resistance in nematodes may be caused by alteration of P-glycoprotein homolog. Molecular and Biochemical Parasitology 91, 327335.CrossRefGoogle Scholar