Abstract
Using laser scanning confocal microscope (LACM) and flow cytometry (FCM), we found that MK571, a specific inhibitor of multidrug resistance-associated protein (MRP), could increase the accumulation of calcein-AM in Prorocentrum lima, a DSP toxin-producing dinoflagellate. This led to the assumption that the cells of P. lima possess MRP transporters in the plasma membrane. The contents of okadaic acid (OA), a main component of DSP toxins, and MRP activities in P. lima were determined at 4, 20, 28, and 42 days after culture and at 38 day under N- and P-limited conditions. We found that both the OA level and MRP activity in P. lima increased over time; OA concentration and MRP activity conformably increased under N- and P-limited conditions compared with control counterparts. These changes in OA concentration and associated increased MRP activity suggest that MRP might be involved in OA transport in P. lima. However, MK571 and CsA, a broad-spectrum inhibitor of multidrug resistance (MDR)-associated ABC proteins, had no observable effects on OA content in P. lima. It seems that except for MDR-associated ABC proteins, other transporters like PDR might be involved in the transport of OA in P. lima.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Andersen P, Throndsen J (2003) Estimating cell numbers. In: Hallegraeff GM, Anderson DM, Cembella AD (eds) Manual on harmful marine microalgae. UNESCO Publishing, Paris, pp 99–129
Barbier M, Amzil Z, Mondeguer F, Bhaud Y, Soyer-Gobillard MO, Lassus P (1999) Okadaic acid and PP2A cellular immunolocalization in Prorocentrum lima (Dinophyceae). Phycologia 38:41–46
Chan LL, Hodgkiss IJ, Lam PKS, Wan JMF, Chou HN, Lum JHK, Lo MGY, Mak ASC, Sit WH, Lo SCL (2005) Use of two-dimensional gel electrophoresis to differentiate morphospecies of Alexandrium minutum, a paralytic shellfish poisoning toxin-producing dinoflagellate of harmful algal blooms. Proteomics 5:1580–1593
Crouzet J, Roland J, Peeters E, Trombik T, Ducos E, Nader J, Boutry M (2013) NtPDR1, a plasma membrane ABC transporter from Nicotiana tabacum, is involved in diterpene transport. Plant Mol Biol 82:181–192
Dean M, Hamon Y, Chimini G (2001) The human ATP-binding cassette (ABC) transporter superfamily. J Lipid Res 42:1007–1017
Espiña B, Louzao MC, Cagide E, Alfonso A, Vieytes MR, Yasumoto T, Botana LM (2010) The methyl ester of okadaic acid is more potent than okadaic acid in disrupting the actin cytoskeleton and metabolism of primary cultured hepatocytes. Brit J Pharmacol 159:337–344
Essodaigui M, Broxterman HJ, Garnier-Suillerot A (1998) Kinetic analysis of calcein and calcein-acetoxymethylester efflux mediated by the multidrug resistance protein and P-glycoprotein. Biochemistry 37:2243–2250
Gekeler V, Ise W, Sanders KH, Ulrich WR, Beck J (1995) The leukotriene LTD4 receptor antagonist MK571 specifically modulates MRP associated multidrug resistance. Biochem Biophys Res Commun 208:345–352
Glibert PM, Burkholder JM, Kana TM (2012) Recent insights about relationships between nutrient availability, forms, and stoichiometry, and the distribution, ecophysiology, and food web effects of pelagic and benthic Prorocentrum species. Harmful Algae 14:231–259
Guillard RL (1975) Culture of phytoplankton for feeding marine invertebrates. In: Smith W, Chanley M (eds) Culture of marine invertebrate animals. Plenum Press, NY, pp 29–60
Hollo Z, Homolya L, Davis CW, Sarkadi B (1994) Calcein accumulation as a fluorometric functional assay of the multidrug transporter. Biochim Biophys Acta 1191:384–388
Huang L, Wang J, Chen WC, Li HY, Liu JS, Jiang T, Yang WD (2014) P-glycoprotein expression in Perna viridis after exposure to Prorocentrum lima, a dinoflagellate producing DSP toxins. Fish Shellfish Immunol 39:254–262
Hung LW, Wang IX, Nikaido K, Liu PQ, Ames GF, Kim SH (1998) Crystal structure of the ATP-binding subunit of an ABC transporter. Nature 396:703–707
Kellmann R, Stuken A, Orr RJS, Svendsen HM, Jakobsen KS (2010) Biosynthesis and molecular genetics of polyketides in marine dinoflagellates. Mar Drugs 8:1011–1048
Lawrence JE, Cembella AD (1999) An immunolabeling technique for the detection of diarrhetic shellfish toxins in individual dinoflagellate cells. Phycologia 38:60–65
Litman T, Druley TE, Stein WD, Bates SE (2001) From MDR to MXR: new understanding of multidrug resistance systems, their properties and clinical significance. Cell Mol Life Sci 58:931–959
Luckenbach T, Epel D (2008) ABCB- and ABCC-type transporters confer multixenobiotic resistance and form an environment-tissue barrier in bivalve gills. Am J Physiol Regul Integr Comp Physiol 294:R1919–R1929
Luedeking A, Van Noorden CJF, Koehler A (2005) Identification and characterisation of a multidrug resistance-related protein mRNA in the blue mussel Mytilus edulis. Mar Ecol Prog Ser 286:167–175
Luo B, Xue XY, Hu WL, Wang LJ, Chen XY (2007) An ABC transporter gene of Arabidopsis thaliana, AtWBC11, is involved in cuticle development and prevention of organ fusion. Plant Cell Physiol 48:1790–1802
McLachlan JL, Marr JC, Conlon-Keily A, Adamson A (1994) Effects of nitrogen concentration and cold temperature on DSP-toxin concentrations in the dinoflagellate Prorocentrum lima (Prorocentrales, Dinophyceae). Nat Toxins 2:263–270
Nascimento SM, Purdie DA, Morris S (2005) Morphology, toxin composition and pigment content of Prorocentrum lima strains isolated from a coastal lagoon in southern UK. Toxicon 45:633–649
Neilan BA, Pearson LA, Muenchhoff J, Moffitt MC, Dittmann E (2013) Environmental conditions that influence toxin biosynthesis in cyanobacteria. Environ Microbiol 15:1239–1253
Nuruzzaman M, Zhang R, Cao HZ, Luo ZY (2014) Plant pleiotropic drug resistance transporters: transport mechanism, gene expression, and function. J Integr Plant Biol 56:729–740
Okumura Y, Koyama J, Takaku H, Satoh H (2001) Influence of organic solvents on the growth of marine microalgae. Arch Environ Contam Toxicol 41:123–128
Olson DP, Taylor BJ, Ivy SP (2001) Detection of MRP functional activity: calcein AM but not BCECF AM as a multidrug resistance-related protein (MRP1) substrate. Cytometry 46:105–113
Pearson LA, Hisbergues M, Borner T, Dittmann E, Neilan BA (2004) Inactivation of an ABC transporter gene, mcyH, results in loss of microcystin production in the cyanobacterium Microcystis aeruginosa PCC 7806. Appl Environ Microbiol 70:6370–6378
Plumley FG (1997) Marine algal toxins: biochemistry, genetics, and molecular biology. Limnol Oceanogr 42:1252–1264
Plumley FG (2001) Purification of an enzyme involved in saxitoxin synthesis. J Phycol 37:926–928
Qadir M, O’Loughlin KL, Fricke SM, Williamson NA, Greco WR, Minderman H, Baer MR (2005) Cyclosporin A is a broad-spectrum multidrug. Resistance modulator. Clin Cancer Res 11:2320–2326
Rea PA (2007) Plant ATP-binding cassette transporters. Annu Rev Plant Biol 58:347–375
Sako Y, Yoshida T, Uchida A, Arakawa O, Noguchi T, Ishida Y (2001) Purification and characterization of a sulfotransferase specific to N-21 of saxitoxin and gonyautoxin 2 + 3 from the toxic dinoflagellate Gymnodinium catenatum (Dinophyceae). J Phycol 37:1044–1051
Scherer C, Wiltshire K, Bickmeyer U (2008) Inhibition of multidrug resistance transporters in the diatom Thalassiosira rotula facilitates dye staining. Plant Physiol Biochem 46:100–103
Schulz B, Kolukisaglu HU (2006) Genomics of plant ABC transporters: the alphabet of photosynthetic life forms or just holes in membranes? FEBS Lett 580:1010–1016
Sharom FJ (2008) ABC multidrug transporters: structure, function and role in chemoresistance. Pharmacogenomics 9:105–127
Sirikantaramas S, Yamazaki M, Saito K (2008a) Mechanisms of resistance to self-produced toxic secondary metabolites in plants. Phytochem Rev 7:467–477
Sirikantaramas S, Yamazaki M, Saito K (2008b) Mutations in topoisomerase I as a self-resistance mechanism coevolved with the production of the anticancer alkaloid camptothecin in plants. Proc Natl Acad Sci U S A 105:6782–6786
Sirikantaramas S, Yamazaki M, Saito K (2009) A survival strategy: the coevolution of the camptothecin biosynthetic pathway and self-resistance mechanism. Phytochemistry 70:1894–1898
Snyder RV, Guerrero MA, Sinigalliano CD, Winshell J, Perez R, Lopez JV, Rein KS (2005) Localization of polyketide synthase encoding genes to the toxic dinoflagellate Karenia brevis. Phytochemistry 66:1767–1780
Stukkens Y, Bultreys A, Grec S, Trombik T, Vanham D, Boutry M (2005) NpPDR1, a pleiotropic drug resistance-type ATP-binding cassette transporter from Nicotiana plumbaginifolia, plays a major role in plant pathogen defense. Plant Physiol 139:341–352
Sugg LM, VanDolah FM (1999) No evidence for an allelopathic role of okadaic acid among ciguatera-associated dinoflagellates. J Phycol 35:93–103
Tillett D, Dittmann E, Erhard M, von Döhren H, Borner T, Neilan BA (2000) Structural organization of microcystin biosynthesis in Microcystis aeruginosa PCC7806: an integrated peptide-polyketide synthetase system. Chem Biol 7:753–764
Vanucci S, Guerrini F, Milandri A, Pistocchi R (2010) Effects of different levels of N- and P-deficiency on cell yield, okadaic acid, DTX-1, protein and carbohydrate dynamics in the benthic dinoflagellate Prorocentrum lima. Harmful Algae 9:590–599
Varkitzi I, Pagou K, Graneli E, Hatzianestis I, Pyrgaki C, Pavlidou A, Montesanto B, Economou-Amilli A (2010) Unbalanced N:P ratios and nutrient stress controlling growth and toxin production of the harmful dinoflagellate Prorocentrum lima (Ehrenberg) Dodge. Harmful Algae 9:304–311
Vilches TS, Norte M, Daranas AH, Fernandez JJ (2012) Biosynthetic studies on water-soluble derivative 5c (DTX5c). Mar Drugs 10:2234–2245
Wang DZ, Dong HP, Li C, Xie ZX, Lin L, Hong HS (2011) Identification and characterization of cell wall proteins of a toxic dinoflagellate Alexandrium catenella using 2-D DIGE and MALDI TOF-TOF mass spectrometry. Evid Based Complement Alternat Med 2011:984080
Wang DZ, Li C, Zhang Y, Wang YY, He ZP, Lin L, Hong HS (2012) Quantitative proteomic analysis of differentially expressed proteins in the toxicity-lost mutant of Alexandrium catenella (Dinophyceae) in the exponential phase. J Proteomics 75:5564–5577
Watanabe T, Kokubu N, Charnick SB, Naito M, Tsuruo T, Cohen D (1997) Interaction of cyclosporin derivatives with the ATPase activity of human P-glycoprotein. Brit J Pharmacol 122:241–248
Xia CQ, Liu N, Miwa GT, Gan LS (2007) Interactions of cyclosporin A with breast cancer resistance protein. Drug Metab Dispos 35:576–582
Yang I, Beszteri S, Tillmann U, Cembella A, John U (2011) Growth- and nutrient-dependent gene expression in the toxigenic marine dinoflagellate Alexandrium minutum. Harmful Algae 12:55–69
Zhou J, Fritz L (1994) Okadaic acid antibody localizes to chloroplasts in the DSP-toxin-producing dinoflagellates Prorocentrum lima and Prorocentrum maculosum. Phycologia 33:455–461
Acknowledgments
This work was supported by the National Natural Science Foundation of China (41230961, 41176088) and the National Basic Research Program of China (973 Program) (2010CB428702).
Author information
Authors and Affiliations
Corresponding authors
Rights and permissions
About this article
Cite this article
Hou, Dy., Liang, Jj., Zou, C. et al. MRP functional activity and character in the dinoflagellate Prorocentrum lima . J Appl Phycol 28, 1667–1676 (2016). https://doi.org/10.1007/s10811-015-0679-1
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10811-015-0679-1