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Marine Drugs 2013

An Overview on the Marine Neurotoxin, Saxitoxin: Genetics, Molecular Targets, Methods of Detection and Ecological Functions

DOI: 10.3390/md11040991

Kathleen D. Cusick,Gary S. Sayler

Keywords: neurotoxin, saxitoxin, ion channels, copper transporter, phytoplankton, paralytic shellfish toxin

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Abstract:

Marine neurotoxins are natural products produced by phytoplankton and select species of invertebrates and fish. These compounds interact with voltage-gated sodium, potassium and calcium channels and modulate the flux of these ions into various cell types. This review provides a summary of marine neurotoxins, including their structures, molecular targets and pharmacologies. Saxitoxin and its derivatives, collectively referred to as paralytic shellfish toxins (PSTs), are unique among neurotoxins in that they are found in both marine and freshwater environments by organisms inhabiting two kingdoms of life. Prokaryotic cyanobacteria are responsible for PST production in freshwater systems, while eukaryotic dinoflagellates are the main producers in marine waters. Bioaccumulation by filter-feeding bivalves and fish and subsequent transfer through the food web results in the potentially fatal human illnesses, paralytic shellfish poisoning and saxitoxin pufferfish poisoning. These illnesses are a result of saxitoxin’s ability to bind to the voltage-gated sodium channel, blocking the passage of nerve impulses and leading to death via respiratory paralysis. Recent advances in saxitoxin research are discussed, including the molecular biology of toxin synthesis, new protein targets, association with metal-binding motifs and methods of detection. The eco-evolutionary role(s) PSTs may serve for phytoplankton species that produce them are also discussed.

References

[1]	Wang, D.-Z. Neurotoxins from marine dinoflagallates: A brief review. Mar. Drugs 2008, 6, 349–371, doi:10.3390/md6020349.
[2]	Rein, K.S.; Borrone, J. Polyketides from dinoflagellates: Origins, pharmacology and biosynthesis. Comp. Biochem. Physiol. B 1999, 124, 117–131, doi:10.1016/S0305-0491(99)00107-8.
[3]	Anderson, P.D. Bioterrorism: Toxins as weapons. J. Pharm. Pract. 2012, 25, 121–129, doi:10.1177/0897190012442351.
[4]	Harada, T.; Oshima, Y.; Yasumoto, T. Studies on paralytic shellfish poisoning in tropical waters: 4. Structures of 2 paralytic shellfish toxins, gonyautoxin-V and gonyautoxin-VI isolated from a tropical dinoflagellate, Pyrodinium bahamense var. compressa. Agric. Biol. Chem. 1982, 46, 1861–1864, doi:10.1271/bbb1961.46.1861.
[5]	Oshima, Y.; Hasegawa, M.; Yasumoto, T.; Hallegraeff, G.; Blackburn, S. Dinoflagellate Gymnodinium catenatum as the source of paralytic shellfish toxins in Tasmanian shellfish. Toxicon 1987, 25, 1105–1111, doi:10.1016/0041-0101(87)90267-4.
[6]	Hallegraeff, G.M.; Steffensen, D.A.; Wetherbee, R. Three estuarine Australian dinoflagellates that can produce paralytic shellfish toxins. J. Plankton Res. 1988, 10, 533–541, doi:10.1093/plankt/10.3.533.
[7]	Anderson, D.M.; Kulis, D.M.; Sullivan, J.J.; Hall, S.; Lee, C. Dynamics and physiology of saxitoxin production by the dinoflagellates Alexandrium spp. Mar. Biol. 1990, 104, 511–524, doi:10.1007/BF01314358.
[8]	Carmichael, W.W.; Evans, W.R.; Yin, Q.Q.; Bell, P.; Moczydlowski, E. Evidence for paralytic shellfish poisons in the freshwater cyanobacterium Lyngbya wollei (Farlow ex Gomont) comb. nov. Appl. Environ. Microbiol. 1997, 63, 3104–3110.
[9]	Negri, A.P.; Jones, G.J. Bioaccumulation of paralytic shellfish poisoning (PSP) toxins from the cyanobacterium Anabaena circinalis by the freshwater mussel Alathyria condola. Toxicon 1995, 33, 667–678, doi:10.1016/0041-0101(94)00180-G.
[10]	Lagos, N.; Onodera, H.; Zagatto, P.A.; Andrinolo, D.; Azevedo, S.M.; Oshima, Y. The first evidence of paralytic shellfish toxins in the freshwater cyanobacterium Cylindrospermopsis raciborskii, isolated from Brazil. Toxicon 1999, 37, 1359–1373, doi:10.1016/S0041-0101(99)00080-X.
[11]	Pomati, F.; Sacchi, S.; Rossetti, C.; Giovannardi, S.; Onodera, H.; Oshima, Y.; Neilan, B.A. The freshwater cyanobacterium Planktothrix sp. FP1: Molecular identification and detection of paralytic shellfish poisoning toxins. J. Phycol. 2000, 36, 553–562, doi:10.1046/j.1529-8817.2000.99181.x.
[12]	Ferreira, F.M.B.; Soler, J.M.F.; Fidalgo, M.L.; Fernandez-Vila, P. PSP toxins from Aphanizomenon flos-aquae (cyanobacteria) collected in the Crestuma-Lever reservoir (Douro river, northern Portugal). Toxicon 2001, 39, 757–761, doi:10.1016/S0041-0101(00)00114-8.
[13]	Shimizu, Y. Microalgal metabolites. Chem. Rev. 1993, 93, 1685–1698, doi:10.1021/cr00021a002.
[14]	Pearson, L.; Mihali, T.; Moffitt, M.; Kellmann, R.; Neilan, B. On the chemistry, toxicology and genetics of the cyanobacterial toxins, microcystin, nodularin, saxitoxin and cylindrospermopsin. Mar. Drugs 2010, 8, 1650–1680, doi:10.3390/md8051650.
[15]	Araoz, R.; Molgo, J.; Tandeau de Marsac, N. Neurotoxic cyanobacterial toxins. Toxicon 2010, 56, 813–828, doi:10.1016/j.toxicon.2009.07.036.
[16]	Schantz, E.J.; Ghazarossian, V.E.; Schnoes, H.K.; Strong, F.M.; Springer, J.P.; Pezzanite, J.O.; Clardy, J. Structure of saxitoxin. J. Am. Chem. Soc. 1975, 97, 1238–1239, doi:10.1021/ja00838a045.
[17]	Rogers, R.S.; Rapoport, H. The pKas of saxitoxin. J. Am. Chem. Soc. 1980, 102, 7335–7339, doi:10.1021/ja00544a030.
[18]	Shimizu, Y.; Hsu, C.P.; Genenah, A. Structure of saxitoxin in solutions and stereochemistry of dihydrosaxitoxins. J. Am. Chem. Soc. 1981, 103, 605–609, doi:10.1021/ja00393a017.
[19]	Schantz, E.J.; Lynch, J.M.; Vayvada, G.; Matsumot, K.; Rapoport, H. Purification and characterization of poison produced by Gonyaulax catenella in axenic culture. Biochemistry 1966, 5, 1191–1195, doi:10.1021/bi00868a011.
[20]	Strichartz, G. Structural determinants of the affinity of saxitoxin for neuronal sodium channels—Electrophysiological studies on frog peripheral nerve. J. Gen. Physiol. 1984, 84, 281–305, doi:10.1085/jgp.84.2.281.
[21]	Genenah, A.A.; Shimizu, Y. Specific toxicity of paralytic shellfish poisons. J. Agric. Food Chem. 1981, 29, 1289–1291, doi:10.1021/jf00108a047.
[22]	Hall, S.; Strichartz, G.; Moczydlowski, E.; Ravindran, A.; Reichardt, P.B. The Saxitoxins: Sources, Chemistry, and Pharmacology. In Marine Toxins: Origin, Structure, and Molecular Pharmacology; Hall, S., Strichartz, G., Eds.; American Chemical Society Symposium Series 418; American Chemical Society: Washington, DC, USA, 1990; pp. 29–65.
[23]	Llewellyn, L.E. Predicitive toxinology: An initial foray using calculated molecular descriptors to decribe toxicity using saxitoxin as a model. Toxicon 2007, 50, 901–913, doi:10.1016/j.toxicon.2007.06.015.
[24]	Cestele, S.; Catterall, W.A. Molecular mechanisms of neurotoxin action on voltage-gated sodium channels. Biochimie 2000, 82, 883–892, doi:10.1016/S0300-9084(00)01174-3.
[25]	Strong, M.; Chandy, K.G.; Gutman, G.A. Molecular evolution of voltage-sensitive ion channel genes: On the origins of electrical excitability. Mol. Biol. Evol. 1993, 10, 221–242.
[26]	Charalambous, K.; Wallace, B.A. NaChBac: The long lost sodium channel ancestor. Biochemistry 2011, 50, 6742–6752, doi:10.1021/bi200942y.
[27]	Stevens, M.; Peigneur, S.; Tytgat, J. Neurotoxins and their binding areas on voltage-gated sodium channels. Front. Pharmacol. 2011, 2, 1–13.
[28]	Catterall, W.A.; Cestele, S.; Yarov-Yarovoy, V.; Yu, F.H.; Konoki, K.; Scheuer, T. Voltage-gated ion channels and gating modifier toxins. Toxicon 2007, 49, 124–141, doi:10.1016/j.toxicon.2006.09.022.
[29]	Catterall, W.A. The Voltage Sensitive Sodium Channel: A Receptor for Multiple Neurotoxins. In Toxic Dinoflagellates; Anderson, D.M., White, A.W., Baden, D.G., Eds.; Elsevier Science Publishing Co., Inc.: New York, NY, USA, 1985; pp. 329–342.
[30]	Noda, M.; Suzuki, H.; Numa, S.; Stuhmer, W. A single point mutation confers tetrodotoxin and saxitoxin insensitivity on the sodium channel II. FEBS Lett. 1989, 259, 213–216, doi:10.1016/0014-5793(89)81531-5.
[31]	Terlau, H.; Heinemann, S.H.; Stuhmer, W.; Pusch, M.; Conti, F.; Imoto, K.; Numa, S. Mapping the site of block by tetrodotoxin and saxitoxin of sodium channel II. FEBS Lett. 1991, 293, 93–96, doi:10.1016/0014-5793(91)81159-6.
[32]	Hartshorne, R.P.; Catterall, W.A. The sodium channel from rat brain—Purification and subunit composition. J. Biol. Chem. 1984, 259, 1667–1675.
[33]	Kao, C.Y.; Walker, S.E. Active groups of saxitoxin and tetrodoxin as deduced from actions of saxitoxin analogs on frog muscle and squid axon. J. Physiol. 1982, 323, 619–637.
[34]	Llewellyn, L.E. Saxitoxin, a toxic marine natural product that targets a multitude of receptors. Nat. Prod. Rep. 2006, 23, 200–222, doi:10.1039/b501296c.
[35]	Baden, D.G.; Trainer, V.L. The Mode and Action of Toxins and Seafood Poisoning. In Algal Toxins in Seafood and Drinking Water; Falconer, I., Ed.; Academic Press: San Diego, CA, USA, 1993.
[36]	Wang, J.X.; Salata, J.J.; Bennett, P.B. Saxitoxin is a gating modifier of hERG K+ channels. J. Gen. Physiol. 2003, 121, 583–598, doi:10.1085/jgp.200308812.
[37]	Murray, S.A.; Mihali, T.K.; Neilan, B.A. Extraordinary conservation, gene loss, and positive selection in the evolution of an ancient neurotoxin. Mol. Biol. Evol. 2011, 28, 1173–1182, doi:10.1093/molbev/msq295.
[38]	Su, Z.; Sheets, M.; Ishida, H.; Li, F.H.; Barry, W.H. Saxitoxin blocks L-type ICa. J. Pharmacol. Exp. Ther. 2004, 308, 324–329.
[39]	Zakon, H.H. Adaptive evolution of voltage-gated sodium channels: The first 800 million years. Proc. Natl. Acad. Sci. USA 2012, 109, 10619–10625, doi:10.1073/pnas.1201884109.
[40]	Mahar, J.; Lukacs, G.L.; Li, Y.; Hall, S.; Moczydlowski, E. Pharmacologiocal and biochemical properties of saxiphilin, a soluble saxitoxin-binding protein from the bullfrog (Rana catesbeiana). Toxicon 1991, 29, 53–71, doi:10.1016/0041-0101(91)90039-T.
[41]	Morabito, M.A.; Moczydlowski, E. Molecular cloning of bullfrog saxiphilin—A unique relative of the transferrin family that binds saxitoxin. Proc. Natl. Acad. Sci. USA 1994, 91, 2478–2482, doi:10.1073/pnas.91.7.2478.
[42]	Gaffney, J.P.; Valentine, A.M. Beyond bilobal: Transferrin homologs having unusual domain architectures. Biochim. Biophys. Acta Gen. Subj. 2012, 1820, 212–217, doi:10.1016/j.bbagen.2011.09.014.
[43]	Morabito, M.A.; Llewellyn, L.E.; Moczydlowski, E.G. Expression of saxiphilin in insect cells and localization of the saxitoxin-binding site to the C-terminal domain homologous to the C-lobe of transferrins. Biochemistry 1995, 34, 13027–13033, doi:10.1021/bi00040a013.
[44]	Llewellyn, L.E.; Moczydlowski, E.G. Characterization of saxitoxin binding to saxiphilin, a relative of the transferrin family that displays pH-dependent ligand-binding. Biochemistry 1994, 33, 12312–12322, doi:10.1021/bi00206a039.
[45]	Cusick, K.D.; Minkin, S.C.; Dodani, S.C.; Chang, C.J.; Wilhelm, S.W.; Sayler, G.S. Inhibition of copper uptake in yeast reveals the copper transporter Ctr1p as a potential molecular target of saxitoxin. Environ. Sci. Technol. 2012, 46, 2959–2966, doi:10.1021/es204027m.
[46]	Hill, K.; Hassett, R.; Kosman, D.; Merchant, S. Regulated copper uptake in Chlamydomonas reinhardtii in response to copper availabilty. Plant. Physiol. 1996, 112, 697–704.
[47]	Dancis, A.; Haile, D.; Yuan, D.S.; Klausner, R.D. The Saccharomyces cerevisiae copper transport protein (Ctr1p). Biochemical characterization, regulation by copper, and physiologic role in copper uptake. J. Biol. Chem. 1994, 269, 25660–25667.
[48]	Dancis, A.; Yuan, D.S.; Haile, D.; Askwith, C.; Eide, D.; Moehle, C.; Kaplan, J.; Klausner, R.D. Molecular characterization of a copper transport protein in S.cerevisiae: An unexpected role for copper in iron transport. Cell 1994, 76, 393–402, doi:10.1016/0092-8674(94)90345-X.
[49]	Page, M.D.; Kropat, J.; Hamel, P.P.; Merchant, S.S. Two Chlamydomonas CTR copper transporters with a novel Cys-Met motif are localized to the plasma membrane and function in copper assimilation. Plant Cell 2009, 21, 928–943, doi:10.1105/tpc.108.064907.
[50]	Aller, S.G.; Unger, V.M. Projection structure of the human copper transporter CTR1 at 6-？ resolution reveals a compact trimer with a novel channel-like architecture. Proc. Natl. Acad. Sci. USA 2006, 103, 3627–3632, doi:10.1073/pnas.0509929103.
[51]	Henderson, R.; Ritchie, J.M.; Strichartz, G.R. Evidence that tetrodotoxin and saxitoxin act at a metal cation binding site in sodium channels of nerve membrane. Proc. Natl. Acad. Sci. USA 1974, 71, 3936–3940, doi:10.1073/pnas.71.10.3936.
[52]	De Feo, C.J.; Aller, S.G.; Unger, V.M. A structural perspective on copper uptake in eukaryotes. Biometals 2007, 20, 705–716, doi:10.1007/s10534-006-9054-7.
[53]	Shimizu, Y. Microalgal metabolites: A new perspective. Annu. Rev. Microbiol. 1996, 50, 431–465, doi:10.1146/annurev.micro.50.1.431.
[54]	Kellmann, R.; Mihali, T.K.; Jeon, Y.J.; Pickford, R.; Pomati, F.; Neilan, B.A. Biosynthetic intermediate analysis and functional homology reveal a saxitoxin gene cluster in cyanobacteria. App. Environ. Microbiol. 2008, 74, 4044–4053, doi:10.1128/AEM.00353-08.
[55]	Kellmann, R.; Neilan, B.A. Biochemical characterization of paralytic shellfish toxin biosynthesis in vitro. J. Phycol. 2007, 43, 497–508, doi:10.1111/j.1529-8817.2007.00351.x.
[56]	Mihali, T.K.; Kellmann, R.; Neilan, B.A. Characterisation of the paralytic shellfish toxin biosynthesis gene clusters in Anabaena circinalis AWQC131C and Aphanizomenon sp. NH-5. BMC Biochem. 2009, 10, 8, doi:10.1186/1471-2091-10-8.
[57]	Mihali, T.K.; Carmichael, W.W.; Neilan, B.A. A putative gene cluster from a Lyngbya wollei bloom that encodes paralytic shellfish toxin biosynthesis. PLoS One 2011, 6, e14657.
[58]	Lin, S.J. Genomic understanding of dinoflagellates. Res. Microbiol. 2011, 162, 551–569, doi:10.1016/j.resmic.2011.04.006.
[59]	Moustafa, A.; Evans, A.N.; Kulis, D.M.; Hackett, J.D.; Erdner, D.L.; Anderson, D.M.; Bhattacharya, D. Transcriptome profiling of a toxic dinoflagellate reveals a gene-rich protist and a potential impact on gene expression due to bacterial presence. PLoS One 2010, 5, e9688, doi:10.1371/journal.pone.0009688.
[60]	Hou, Y.; Lin, S. Distinct gene number-genome size relationships for eukaryotes and non-eukaryotes: Gene content estimation for dinoflagellate genomes. PLoSOne 2009, 4, e6978.
[61]	Erdner, D.L.; Anderson, D.M. Global transcriptional profiling of the toxic dinoflagellate Alexandrium fundyense using massively parallel signature sequencing. BMC Genomics 2006, 7, 88, doi:10.1186/1471-2164-7-88.
[62]	Stuken, A.; Orr, R.J.S.; Kellmann, R.; Murray, S.A.; Neilan, B.A.; Jakobsen, K.S. Discovery of nuclear-encoded genes for the neurotoxin saxitoxin in dinoflagellates. PLoS One 2011, 6, e20096.
[63]	Hackett, J.D.; Wisecarver, J.H.; Brosnahan, M.L.; Kulis, D.M.; Anderson, D.M.; Bhattacharya, D.; Plumley, F.G.; Erdner, D.L. Evolution of saxitoxin synthesis in cyanobacteria and dinoflagellates. Mol. Biol. Evol. 2013, 30, 70–78, doi:10.1093/molbev/mss142.
[64]	Bachvaroff, T.R.; Place, A.R. From stop to start: Tandem gene arrangement, copy number and trans-splicing sites in the dinoflagellate Amphidinium carterae. PLoS One 2008, 3, e2929, doi:10.1371/journal.pone.0002929.
[65]	Le, Q.H.; Markovic, P.; Hastings, J.W.; Jovine, R.V.M.; Morse, D. Structure and organization of the peridinin chlorophyll a binding protein gene in Gonyaulax polyedra. Mol. Gen. Genet. 1997, 255, 595–604, doi:10.1007/s004380050533.
[66]	Moustafa, A.; Loram, J.E.; Hackett, J.D.; Anderson, D.M.; Plumley, F.G.; Bhattacharya, D. Origin of saxitoxin biosynthetic genes in cyanobacteria. PLoS One 2009, 4, e5758.
[67]	Salcedo, T.; Upadhyay, R.J.; Nagasaki, K.; Bhattacharya, D. Dozens of toxin-related genes are expressed in a nontoxic strain of the dinoflagellate Heterocapsa circularisquama. Mol. Biol. Evol. 2012, 29, 1503–1506, doi:10.1093/molbev/mss007.
[68]	Yoshida, T.; Sako, Y.; Uchida, A.; Kakutani, T.; Arakawa, O.; Noguchi, T.; Ishida, Y. Purification and characterization of sulfotransferase specific to O-22 of 11-hydroxy saxitoxin from the toxic dinoflagellate Gymnodinium catenatum (Dinophyceae). Fish. Sci. 2002, 68, 634–642, doi:10.1046/j.1444-2906.2002.00471.x.
[69]	Sako, Y.; Yoshida, T.; Uchida, A.; Arakawa, O.; Noguchi, T.; Ishida, Y. 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. 2001, 37, 1044–1051, doi:10.1046/j.1529-8817.2001.00119.x.
[70]	Landsberg, J.H.; Hall, S.; Johannessen, J.N.; White, K.D.; Conrad, S.M.; Abbott, J.P.; Flewelling, L.J.; Richardson, R.W.; Dickey, R.W.; Jester, E.L.E.; et al. Saxitoxin puffer fish poisoning in the United States, with the first report of Pyrodinium bahamense as the putative toxin source. Environ. Health Perspect. 2006, 114, 1502–1507, doi:10.1289/ehp.8998.
[71]	Etheridge, S.M. Paralytic shellfish poisoning: Seafood safety and human health perspectives. Toxicon 2010, 56, 108–122.
[72]	Deeds, J.R.; Landsberg, J.H.; Etheridge, S.M.; Pitcher, G.C.; Longan, S.W. Non-traditional vectors for paralytic shellfish poisoning. Mar. Drugs 2008, 6, 308–348, doi:10.3390/md6020308.
[73]	Van Dolah, F.M. Marine algal toxins: Origins, health effects, and their increased occurrence. Environ. Health Perspect. 2000, 108, 133–141, doi:10.1289/ehp.00108s1133.
[74]	Landsberg, J.H. The effects of harmful algal blooms on aquatic organisms. Rev. Fish. Sci. 2002, 10, 113–390.
[75]	Abbott, J.P.; Flewelling, L.J.; Landsberg, J.H. Saxitoxin monitoring in three species of Florida puffer fish. Harmful Algae 2009, 8, 343–348, doi:10.1016/j.hal.2008.07.005.
[76]	Tamplin, M.L. A Bacterial Source of Tetrodotoxins and Saxitoxins. In Marine Toxins: Origin, Structure, and Molecular Pharmacology; Hall, S.L., Strichartz, G., Eds.; American Chemical Society: Washington, DC, USA, 1990; pp. 78–86.
[77]	Schantz, E.J.; McFarren, E.F.; Schafer, M.L.; Lewis, K.H. Purified shellfsih poison for bioassay standardization. J. Assoc. Off. Anal. Chem. 1958, 41, 160–168.
[78]	Aune, T.; Ramstad, H.; Heidenreich, B.; Landsverk, T.; Waaler, T.; Egaas, E.; Julshamn, K. Zinc accumulation in oysters giving mouse deaths in paralytic shellfish poisoning bioassay. J. Shellfish Res. 1998, 17, 1243–1246.
[79]	Turner, A.D.; Dhanji-Rapkova, M.; Algoet, M.; Suarez-Isla, B.A.; Cordova, M.; Caceres, C.; Murphy, C.J.; Casey, M.; Lees, D.N. Investigations into matrix components affecting the performance of the official bioassay reference method for quantitation of paralytic shellfish poisoning toxins in oysters. Toxicon 2012, 59, 215–230.
[80]	Aune, T.; Aasen, J.A.B.; Miles, C.O.; Larsen, S. Effect of mouse strain and gender on LD50 of yessotoxin. Toxicon 2008, 52, 535–540.
[81]	Food and Agricultural Organization. Marine Biotoxins FAO Food and Nutrition Paper 80; Food and Agricultural Organization of the United Nations: Rome, Italy, 2004.
[82]	Humpage, A.R.; Magalhaes, V.F.; Froscio, S.M. Comparison of analytical tools and biological assays for detection of paralytic shellfish poisoning toxins. Anal. Bioanal. Chem. 2010, 397, 1655–1671, doi:10.1007/s00216-010-3459-4.
[83]	Louzao, M.C.; Vieytes, M.R.; Baptista de Sousa, J.M.V.; Leira, F.; Botana, L.M. A fluorometric method based on changes in membrane potential for screening paralytic shellfish toxins in mussels. Anal. Biochem. 2001, 289, 246–250, doi:10.1006/abio.2000.4942.
[84]	Cheng, J.P.; Pi, S.S.; Ye, S.F.; Gao, H.M.; Yao, L.; Jiang, Z.Y.; Song, Y.L.; Xi, L. A new simple screening method for the detection of paralytic shellfish poisoning toxins. Chin. J. Ocean. Limnol. 2012, 30, 786–790, doi:10.1007/s00343-012-1097-8.
[85]	Sullivan, J.J. High-Performance Liquid Chromatographic Method Applied to Paralytic Shellfish Poisoning Research. In Marine Toxins: Origin, Structure, and Molecular Pharmacology; Hall, S., Strichartz, G., Eds.; American Chemical Society: Washington, DC, USA, 1990; pp. 66–77.
[86]	Turner, A.D.; Hatfield, R.C. Refinement of AOAC official method (SM) 2005.06 liquid chromatography-fluorescence detection method to improve performance characteristics for the determination of paralytic shellfish toxins in king and queen scallops. J. AOAC Int. 2012, 95, 129–142, doi:10.5740/jaoacint.11-184.
[87]	Dell’Aversano, C.; Eaglesham, G.K.; Quilliam, M.A. Analysis of cyanobacterial toxins by hydrophilic interaction liquid chromatography-mass spectrometry. J. Chromatogr. A 2004, 1028, 155–164, doi:10.1016/j.chroma.2003.11.083.
[88]	Dell’Aversano, C.; Hess, P.; Quilliam, M.A. Hydrophilic interaction liquid chromatography-mass spectrometry for the analysis of paralytic shellfish poisoning (PSP) toxins. J. Chromatogr. A 2005, 1081, 190–201, doi:10.1016/j.chroma.2005.05.056.
[89]	Halme, M.; Rapinoja, M.L.; Karjalainen, M.; Vanninen, P. Verification and quantification of saxitoxin from algal samples using fast and validated hydrophilic interaction liquid chromatography-tandem mass spectrometry method. J. Chromatogr. B 2012, 880, 50–57, doi:10.1016/j.jchromb.2011.11.015.
[90]	Davio, S.R.; Fontelo, P.A. A competitive displacement assay to detect saxitoxin and tetrodotoxin. Anal. Biochem. 1984, 141, 199–204, doi:10.1016/0003-2697(84)90446-9.
[91]	Vieytes, M.R.; Cabado, A.G.; Alfonso, A.; Louzao, M.C.; Botana, A.M.; Botana, L.M. Solid-phase radioreceptor assay for paralytic shellfish toxins. Anal. Biochem. 1993, 211, 87–93, doi:10.1006/abio.1993.1237.
[92]	Doucette, G.J.; Logan, M.M.; Ramsdell, J.S.; van Dolah, F.M. Development and preliminary validation of a microtiter plate-based receptor binding assay for paralytic shellfish poisoning toxins. Toxicon 1997, 35, 625–636, doi:10.1016/S0041-0101(96)00189-4.
[93]	Usup, G.; Leaw, C.-P.; Cheah, M.-Y.; Ahmad, A.; Ng, B.-K. Analysis of paralytic shellfish poisoning congeners by a sodium channel receptor binding assay. Toxicon 2004, 44, 37–43, doi:10.1016/j.toxicon.2004.03.026.
[94]	Van Dolan, F.M.; Fire, S.E.; Leighfield, T.A.; Mikulski, C.M.; Doucette, G.J. Determination of paralytic shellfish toxins in shellfish by Receptor Binding Assay: Collaborative study. J. AOAC Int. 2012, 95, 795–812, doi:10.5740/jaoacint.CS2011_27.
[95]	Kogure, K.; Tamplin, M.L.; Simidu, U.; Colwell, R.R. A tissue culture assay for tetrodotoxin, saxitoxin, and related toxins. Toxicon 1988, 26, 191–197, doi:10.1016/0041-0101(88)90171-7.
[96]	Jellett, J.F.; Marks, L.J.; Stewart, J.E.; Dorey, M.I.; Watson-Wright, W.; Lawrence, J.F. Paralytic shellfish poison (saxitoxin family) bioassays: Automated endpoint determination and stadardization of the in vitro tissue culture bioassay, and comparison with the standard mouse bioassay. Toxicon 1992, 30, 1143–1156, doi:10.1016/0041-0101(92)90430-D.
[97]	Gallacher, S.; Birkbeck, T.H. A tissue culture assay for direct detection of sodium channel blocking toxins in bacterial culture supernatents. FEMS Microbiol. Lett. 1992, 92, 101–108, doi:10.1016/0378-1097(92)90549-4.
[98]	Manger, M.L.; Lega, L.S.; Lee, S.Y.; Hungerford, J.M.; Wekell, M.M. Tetrazolium-based bioassay for neurotoxins active on voltage-sensitive sodium channels: Semiautomated assay for saxitoxins, brevetoxins, and ciguatoxins. Anal. Biochem. 1993, 214, 190–194, doi:10.1006/abio.1993.1476.
[99]	Jellett, J.F.; Doucette, L.I.; Belland, E.R. The MIST (TM) shipable cell bioassay kits for PSP: An alternative to the mouse bioassay. J. Shellfish Res. 1998, 17, 1653–1655.
[100]	Nicholson, R.A.; Li, G.H.; Buenaventura, E.; Graham, D. A rapid and sensitive assay for paralytic shellfish poison (PSP) toxins using mouse brain synaptoneurosomes. Toxicon 2002, 40, 831–838, doi:10.1016/S0041-0101(02)00083-1.
[101]	Louzao, M.C.; Vieytes, M.R.; Cabado, A.G.; de Sousa, J.; Botana, L.M. A fluorimetric microplate assay for detection and quantitation of toxins causing paralytic shellfish poisoning. Chem. Res. Toxicol. 2003, 16, 433–438, doi:10.1021/tx025574r.
[102]	Manger, R.; Woodle, D.; Berger, A.; Hungerford, J. Flow cytometric detection of saxitoxins using fluorescent voltage-sensitive dyes. Anal. Biochem. 2007, 366, 149–155, doi:10.1016/j.ab.2007.04.010.
[103]	Usleber, E.; Schneider, E.; Terplan, G. Direct enzyme immunoassay in microtitration plate and test strip format for the detection of saxitoxin in shellfish. Lett. Appl. Microbiol. 1991, 13, 275–277, doi:10.1111/j.1472-765X.1991.tb00627.x.
[104]	Chu, F.S.; Huang, X.; Hall, S. Production and characterization of antibodies against neosaxitoxin. J. AOAC Int. 1992, 75, 341–345.
[105]	Burk, C.; Usleber, E.; Dietrich, R.; Martlbauer, E. Production and characterization of antibodies against neosaxitoxin utilizing a novel immunogen synthesis procedure. Food Agric. Immunol. 1995, 7, 315–322, doi:10.1080/09540109509354891.
[106]	Micheli, L.; di Stefano, S.; Moscone, D.; Palleschi, G.; Marini, S.; Coletta, M.; Draisci, R.; Quadri, F.D. Production of antibodies and development of highly sensitive formats of enzyme immunoassay for saxitoxin analysis. Anal. Bioanal. Chem. 2002, 373, 678–684, doi:10.1007/s00216-002-1399-3.
[107]	Huang, X.; Hsu, K.H.; Chu, F.S. Direct competitive enzyme-linked immunosorbent assay for saxitoxin and neosaxitoxin. J. Agric. Food Chem. 1996, 44, 1029–1035, doi:10.1021/jf950717j.
[108]	Kawatsu, K.; Hamano, Y.; Sugiyama, A.; Hashizume, K.; Noguchi, T. Development and application of an enzyme immunoassay based on a monoclonal antibody against gonyautoxin components of paralytic shellfish poisoning toxins. J. Food Prot. 2002, 65, 1304–1308.
[109]	Dubois, M.; Demoulin, L.; Charlier, C.; Singh, G.; Godefroy, S.B.; Campbell, K.; Elliott, C.T.; Delahaut, P. Development of ELISAs for detecting domoic acid, okadaic acid, and saxitoxin and their applicability for the detection of marine toxins in samples collected in Belgium. Food Addit. Contam. Part A 2010, 27, 859–868, doi:10.1080/19440041003662881.
[110]	Garneau, M.E.; Schnetzer, A.; Countway, P.D.; Jones, A.C.; Seubert, E.L.; Caron, D.A. Examination of the seasonal dynamics of the toxic dinoflagellate Alexandrium catenella at Redondo Beach, California, by quantitative PCR. Appl. Environ. Microbiol. 2011, 77, 7669–7680.
[111]	Murray, S.A.; Wiese, M.; Stuken, A.; Brett, S.; Kellmann, R.; Hallegraeff, G.; Neilan, B.A. SxtA-based quantitative molecular assay to identify saxitoxin-producing harmful algal blooms in marine waters. Appl. Environ. Microbiol. 2011, 77, 7050–7057, doi:10.1128/AEM.05308-11.
[112]	Al-Tebrineh, J.; Pearson, L.A.; Yasar, S.A.; Neilan, B.A. A multiplex qPCR targeting hepato- and neurotoxigenic cyanobacteria of global significance. Harmful Algae 2012, 15, 19–25, doi:10.1016/j.hal.2011.11.001.
[113]	Nagai, S.; Itakura, S. Specific detection of the toxic dinoflagellates Alexandrium tamarense and Alexandrium catenella from single vegetative cells by a loop-mediated isothermal amplification method. Mar. Genomics 2012, 7, 43–49, doi:10.1016/j.margen.2012.03.001.
[114]	Notomi, T.; Okayama, H.; Masubuchi, H.; Yonezawa, T.; Watanabe, K.; Amino, N.; Hase, T. Loop-mediated isothermal amplification of DNA. Nucl. Acids Res. 2000, 28, 63, doi:10.1093/nar/28.12.e63.
[115]	Mori, Y.; Nagamine, K.; Tomita, N.; Notomi, T. Detection of loop-mediated isothermal amplification reaction by turbidity derived from magnesium pyrophosphate formation. Biochem. Biophys. Res. Commun. 2001, 289, 150–154, doi:10.1006/bbrc.2001.5921.
[116]	Enosawa, M.; Kageyama, S.; Sawai, K.; Watanabe, K.; Notomi, T.; Onoe, S.; Mori, Y.; Yokomizo, Y. Use of loop-mediated isothermal amplification of the IS900 sequence for rapid detection of cultured Mycobacterium avium subsp. paratuberculosis. J. Clin. Microbiol. 2003, 41, 4359–4365, doi:10.1128/JCM.41.9.4359-4365.2003.
[117]	Tang, X.H.; Yu, R.C.; Zhou, M.J.; Yu, Z.G. Application of rRNA probes and fluorescence in situ hybridization for rapid detection of the toxic dinoflagellate Alexandrium minutum. Chin. J. Ocean. Limnol. 2012, 30, 256–263, doi:10.1007/s00343-012-1142-7.
[118]	Rhodes, L.; Smith, K.; De Salas, M. DNA probes, targeting large sub-unit rRNA, for the rapid identification of the paralytic shellfish poison producing dinoflagellate, Gymnodinium catenatum. New Z. J. Mar. Freshw. Res. 2007, 41, 385–390, doi:10.1080/00288330709509928.
[119]	Wyatt, T.; Jenkinson, I.R. Notes on Alexandrium population dynamics. J. Plankton Res. 1997, 19, 551–575, doi:10.1093/plankt/19.5.551.
[120]	Ritson-Williams, R.; Yotsu-Yamashita, M.; Paul, V.J. Ecological functions of tetrodotoxin in a deadly polyclad flatworm. Proc. Natl. Acad. Sci. USA 2006, 103, 3176–3179, doi:10.1073/pnas.0506093103.
[121]	Cembella, A.D. Chemical ecology of eukaryotic microalgae in marine ecosystems. Phycologia 2003, 42, 420–447, doi:10.2216/i0031-8884-42-4-420.1.
[122]	Anderson, D.M.; Cheng, T.P.-O. Intracellular localization of saxitoxins in the dinofllagellate Gonyaulax tamarensis. J. Phycol. 1988, 24, 17–22, doi:10.1111/j.1529-8817.1988.tb04451.x.
[123]	Cembella, A.D. Ecophysiology and Metabolism of Paralytic Shellfish Toxins in Marine Microalgae. In Physiological Ecology of Harmful Algal Blooms; Anderson, D.M., Cembella, A.D., Hallegraeff, G., Eds.; Springer-Verlag: Berlin, Germany, 1998; pp. 381–403.
[124]	Zimmer, R.K.; Ferrer, R.P. Neuroecology, chemical defense, and the keystone species concept. Biol. Bull. 2007, 213, 208–225, doi:10.2307/25066641.
[125]	Oikawa, H.; Satomi, M.; Watabe, S.; Yano, Y. Accumulation and depuration rates of paralytic shellfish poisoning toxins in the shore crab Telmessus acutidens by feeding toxic mussels under laboratory controlled conditions. Toxicon 2005, 45, 163–169, doi:10.1016/j.toxicon.2004.10.004.
[126]	Robineau, B.; Gagné, J.A.; Fortier, L.; Cembella, A.D. Potential impact of a toxic dinoflagellate (Alexandrium excavatum) bloom on survival of fish and crustacean larvae. Mar. Biol. 1991, 108, 293–301, doi:10.1007/BF01344344.
[127]	Selander, E.; Thor, P.; Toth, G.; Pavia, H. Copepods induce paralytic shellfish toxin production in marine dinoflagellates. Proc. R. Soc. B 2006, 273, 1673–1680, doi:10.1098/rspb.2006.3502.
[128]	Bergkvist, J.; Selander, E.; Pavia, H. Induction of toxin production in dinoflagellates: The grazer makes a difference. Oecologia 2008, 156, 147–154, doi:10.1007/s00442-008-0981-6.
[129]	Teegarden, G.J. Copepod grazing selection and particle discrimination on the basis of PSP toxin content. Mar. Ecol. Prog. Ser. 1999, 181, 163–176, doi:10.3354/meps181163.
[130]	Da Costa, R.M.; Franco, J.; Cacho, E.; Fernandez, F. Toxin content and toxic effects of the dinoflagellate Gyrodinium corsicum (Paulmier) on the ingestion and survival rates of the copepods Acartia grani and Euterpina acutifrons. J. Exp. Mar. Biol. Ecol. 2005, 322, 177–183, doi:10.1016/j.jembe.2005.02.017.
[131]	Frangoulos, M.; Guisande, C.; Maneiro, I.; Riveiro, I.; Franco, J. Short-term and long-term effects of the toxic dinoflagellate Alexandrium minutum on the copepod Acartia clausi. Mar. Ecol. Prog. Ser. 2000, 203, 161–169, doi:10.3354/meps203161.
[132]	Barreiro, A.; Guisande, C.; Frangopulos, M.; Gonzalez-Fernandez, A.; Munoz, S.; Perez, D.; Magadan, S.; Maneiro, I.; Riveiro, I.; Iglesias, P. Feeding strategies of the copepod Acartia clausi on single and mixed diets of toxic and non-toxic strains of the dinoflagellate Alexandrium minutum. Mar. Ecol. Prog. Ser. 2006, 316, 115–125, doi:10.3354/meps316115.
[133]	Turner, J.T.; Tester, P.A. Toxic marine phytoplankton, zooplankton grazers, and pelagic food webs. Limnol. Oceanogr. 1997, 42, 1203–1214, doi:10.4319/lo.1997.42.5_part_2.1203.
[134]	Solé, J.; Estrada, M.; Garcia-Ladona, E. Biological control of harmful algal blooms: A modelling study. J. Mar. Syst. 2006, 61, 165–179, doi:10.1016/j.jmarsys.2005.06.004.
[135]	Tillmann, U.; John, U. Toxic effects of Alexandrium spp. on heterotrophic dinoflagellates: An allelochemical defence mechanism independent of PSP-toxin content. Mar. Ecol. Prog. Ser. 2006, 61, 47–59.
[136]	Juhl, A.R.; Martins, C.A.; Anderson, D.M. Toxicity of Alexandrium lusitanicum to gastropod larvae is not caused by paralytic-shellfish-poisoning toxins. Harmful Algae 2008, 7, 567–573, doi:10.1016/j.hal.2007.12.019.
[137]	Ianora, A.; Bentley, M.G.; Caldwell, G.S.; Casotti, R.; Cembella, A.D.; Engstrom-Ost, J.; Halsband, C.; Sonnenschein, E.; Legrand, C.; Llewellyn, C.A.; et al. The relevance of marine chemical ecology to plankton and ecosystem function: An emerging field. Mar. Drugs 2011, 9, 1625–1648.

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