1887

Abstract

Saxitoxin (STX) is the most potent representative among the paralytic shellfish poisoning (PSP) toxins, which are highly selective Na channel-blocking alkaloids. This study investigated, in cultures of the cyanobacterium T3, the effects of pH, salt, amiloride and lidocaine hydrochloride on total cellular levels of Na and K ions and STX accumulation. Both Na levels and intracellular STX concentrations increased exponentially in response to rising alkalinity. NaCl inhibited cyanobacterial growth at a concentration of 10 mM. In comparison with osmotically stressed controls, however, NaCl promoted STX accumulation in a dose-dependent manner. A correlation was seen in the time-course of both total cellular Na levels and intracellular STX for NaCl, amiloride and lidocaine exposure. The increase in cellular Na induced by NaCl at 10 mM was coupled with a proportional accumulation of STX. The two Na channel-blocking agents amiloride and lidocaine had opposing effects on both cellular Na levels and STX accumulation. Amiloride at 1 mM reduced ion and toxin concentrations, while lidocaine at 1 μM increased the total cellular Na and STX levels. The effects of the channel-blockers were antagonistic and dependent on an alkaline pH. The results presented suggest that, in T3, STX is responsive to cellular Na levels. This may indicate that either STX metabolism or the toxin itself could be linked to the maintenance of cyanobacterial homeostasis. The results also enhance the understanding of STX production and the ecology of PSP toxin-producing cyanobacteria.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.26350-0
2004-02-01
2020-10-28
Loading full text...

Full text loading...

/deliver/fulltext/micro/150/2/mic1500455.html?itemId=/content/journal/micro/10.1099/mic.0.26350-0&mimeType=html&fmt=ahah

References

  1. Alam M., Ikawa M., Sasner J. J. Jr, Sawyer P. J. 1973; Purification of Aphanizomenon flos-aquae toxin and its chemical and physiological properties. Toxicon11:65–72[CrossRef]
    [Google Scholar]
  2. Ali R. M. 2000; Role of putrescine in salt tolerance of Atropa belladonna plant. Plant Science152:173–179[CrossRef]
    [Google Scholar]
  3. Apte S. K., Thomas J. 1986; Membrane electrogenesis and sodium transport in filamentous nitrogen-fixing cyanobacteria. Eur J Biochem154:395–401[CrossRef]
    [Google Scholar]
  4. Bidani A., Heming T. A. 1997; Effects of lidocaine on cytosolic pH regulation and stimulus-induced effector functions in alveolar macrophages. Lung175:349–361[CrossRef]
    [Google Scholar]
  5. Bowling L. C., Baker P. D. 1996; Major cyanobacterial bloom in the Barwon-Darling River, Australia, in 1991, and underlying limnological conditions. Mar Freshw Res47:643–657[CrossRef]
    [Google Scholar]
  6. Bradford M. M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem72:248–254[CrossRef]
    [Google Scholar]
  7. Carmichael W. W., Evans W. R., Yin Q. Q., Bell P., Moczydlowsky E. 1997; Evidence of paralytic shellfish poisons in the freshwater cyanobacterium Lyngbya wollei (Farlow ex Gomont) comb. nov. Appl Environ Microbiol63:3104–3110
    [Google Scholar]
  8. Catteral W. A., Morrow G. S., Hartsharne R. P. 1979; Neurotoxin binding to receptor sites associated with the voltage sensitive sodium channels in intact, lysed, and detergent solubilized brain membranes. J Biol Chem254:11379–11387
    [Google Scholar]
  9. Catterall W. A. 1980; Neurotoxins that act on voltage-sensitive sodium channels in excitable membranes. Annu Rev Pharmacol Toxicol20:15–43[CrossRef]
    [Google Scholar]
  10. Cestèle S., Catterall W. A. 2000; Molecular mechanisms of neurotoxin action on voltage-gated sodium channels. Biochimie82:883–892[CrossRef]
    [Google Scholar]
  11. Gallacher S., Smith E. A. 1999; Bacteria and paralytic shellfish toxins. Protist150:245–255[CrossRef]
    [Google Scholar]
  12. Gorham P. R., McLachlan J., Hammer U. T., Kim W. K. 1964; Isolation and culture of toxic strains of Anabaena flos-aquae (Lyngb.) de Bréb. Verh Int Verein Theor Angew Limnol15:796–804
    [Google Scholar]
  13. Harada T., Oshima Y., Yasumoto T. 1982; Structure of two paralytic shellfish toxins, gonyautoxins V and VI, isolated from a tropical dinoflagellate, Pyrodinium bahamense var. compressa . Agric Biol Chem46:1861–1864[CrossRef]
    [Google Scholar]
  14. Hille B. 1997; Ionic Channels of Excitable Membranes , 2nd edn. Sunderland, MA: Sinauer Associates;
    [Google Scholar]
  15. Horikoshi K. 1991; Microorganisms in Alkaline Environments New York: VCH;
    [Google Scholar]
  16. Humpage A. R., Rositano J., Bretag A., Brown R., Baker P., Nicholson B. C., Steffensen D. A. 1994; Paralytic shellfish poisons from Australian cyanobacterial blooms. Aust J Mar Freshw Res45:761–771[CrossRef]
    [Google Scholar]
  17. Hwang D. F., Lu Y. H. 2000; Influence of environmental and nutritional factors on growth, toxicity, and toxin profile of dinoflagellate Alexandrium minutum . Toxicon38:1491–1503[CrossRef]
    [Google Scholar]
  18. Kaas H., Henriksen P. 2000; Saxitoxins (PSP toxins) in Danish lakes. Water Res34:2089–2097[CrossRef]
    [Google Scholar]
  19. Kim D., Smith T. W. 1986; Effects of amiloride and ouabain on contractile state, Ca and Na fluxes, and Na content in cultured chick heart cells. Mol Pharmacol29:363–371
    [Google Scholar]
  20. Krulwich T. A., Ito M., Guffanti A. A. 2001; The Na+-dependence of alkaliphily in Bacillus . Biochim Biophys Acta 1505;158–168[CrossRef]
    [Google Scholar]
  21. Lagos N., Onodera H., Zagatto P. A., Andrinolo D., Azevedo S. M. F. Q., Oshima Y. 1999; The first evidence of paralytic shellfish toxins in the freshwater cyanobacterium Cylindrospermopsis raciborskii , isolated from Brazil. Toxicon37:1359–1373[CrossRef]
    [Google Scholar]
  22. Lawrence J. F., Wong B., Ménard C. 1996; Determination of decarbamoyl saxitoxin and its analogues in shellfish by prechromatographic oxidation and liquid chromatography with fluorescence detection. J AOAC Int79:1111–1116
    [Google Scholar]
  23. Lengeler J. W., Drews G., Schlegel H. G. (editors) 1999; Biology of the Prokaryotes Oxford: Blackwell Science;
    [Google Scholar]
  24. Maestri O., Joset F. 2000; Regulation by external pH and stationary growth phase of the acetolactate synthase from Synechocystis PCC6803. Mol Microbiol37:828–838[CrossRef]
    [Google Scholar]
  25. Miller A. G., Turpin D. H., Canvin D. T. 1984; Na+ requirement for growth, photosynthesis, and pH regulation in the alkalotolerant cyanobacterium Synechococcus leopoliensis . J Bacteriol159:100–106
    [Google Scholar]
  26. Negri A. P., Jones G. J., Blackburn S., Oshima Y., Onodera H. 1997; Effect of culture and bloom development, and of sample storage on paralytic shellfish poisons in the cyanobacterium Anabaena circinalis . J Phycol33:85–98
    [Google Scholar]
  27. Nosaka S., Ohkawa T. A., Okihara K., Yoshikawa K. 1992; Effects of local anesthetics on the Chara plasmalemma. Biochim Biophys Acta1106:325–334[CrossRef]
    [Google Scholar]
  28. Ogata T., Kodama M., Ishimara T. 1987; Toxin production in the dinoflagellate Protogonyaulax tamarensis . Toxicon25:923–928[CrossRef]
    [Google Scholar]
  29. Pereira P., Onodera H., Andrinolo D., Franca S., Araujo F., Lagos N., Oshima Y. 2000; Paralytic shellfish toxins in the freshwater cyanobacterium Aphanizomenon flos-aquae , isolated from Montargil reservoir. Portugal. Toxicon38:1689–1702[CrossRef]
    [Google Scholar]
  30. Pomati F., Sacchi S., Rossetti C., Giovannardi S., Onodera H., Oshima Y., Neilan B. A. 2000; The freshwater cyanobacterium Planktothrix sp. FP1: molecular identification and detection of paralytic shellfish poisoning toxins. J Phycol36:553–562[CrossRef]
    [Google Scholar]
  31. Pomati F., Neilan B. A., Manarolla G., Suzuki T., Rossetti C. 2003a; Enhancement of intracellular saxitoxin accumulation by lidocaine hydrochloride in the cyanobacterium Cylindrospermopsis raciborskii T3 ( Nostocales ). J Phycol39:535–542[CrossRef]
    [Google Scholar]
  32. Pomati F., Rossetti C., Calamari D., Neilan B. A. 2003b; Effects of saxitoxin (STX) and veratridine on bacterial Na+/K+ fluxes: a prokaryote-based STX bioassay. Appl Environ Microbiol69:7371–7376[CrossRef]
    [Google Scholar]
  33. Quintero M. J., Muro-Pastor A. M., Herrero A., Flores E. 2000; Arginine catabolism in the cyanobacterium Synechocystis sp. strain PCC6803 involves the urea cycle and arginase pathway. J Bacteriol182:1008–1015[CrossRef]
    [Google Scholar]
  34. Rowbury R. J., Goodson M., Humphrey T. J. 1994; Sodium chloride induces an NhaA/NhaR-independent acid sensitivity at neutral external pH in Escherichia coli . Appl Environ Microbiol60:1630–1634
    [Google Scholar]
  35. Schaefer L., Sakai H., Mattei M. G., Lazdunski M., Lingueglia E. 2001; Molecular cloning, functional expression and chromosomal localization of an amiloride-sensitive Na+ channel from human small intestine. FEBS Lett471:205–210
    [Google Scholar]
  36. Shibamoto S., Hori T., Hayakawa M., Nango M., Oku N., Ito F, Cragoe E. J. Jr. 1990; Inhibitory effect of local anesthetics on Na+/H+ antiporter in brush border membrane-reconstituted vesicles. Life Sci47:1129–1133[CrossRef]
    [Google Scholar]
  37. Shimizu Y. 1977; Chemistry and distribution of deleterious dinoflagellate toxins. In Marine Natural Products Chemistry pp261–269 Edited by Faulkner D. J., Fenical W. H.. New York: Plenum;
    [Google Scholar]
  38. Shimizu Y. 1996; Microalgal metabolites – a new perspective. Annu Rev Microbiol50:431–465[CrossRef]
    [Google Scholar]
  39. Sivonen K., Jones G. 1999; Cyanobacterial toxins. In Toxic Cyanobacteria in Water pp41–111 Edited by Chorus I., Bartram J.. London: E. & F. N. Spon;
    [Google Scholar]
  40. Sonoda M., Katoh H., Vermaas W., Schmetterer G., Ogawa T. 1998; Photosynthetic electron transport involved in PxcA-dependent proton extrusion in Synechocystis sp. strain PCC6803: effect of pxcA inactivation on CO2, HCO3, and NO3 uptake. J Bacteriol180:3799–3803
    [Google Scholar]
  41. Strichartz G. 1981; Relative potencies of several derivates of saxitoxin: electrophysiological and toxin binding studies. Biophys J33:209–216
    [Google Scholar]
  42. Suzuki T., Ezure T., Yamaguchi T., Domen H., Ishida M., Schmidt W. 2000; Stimulatory effect of procaine on the growth of several microalgae and cyanobacteria. J Pharm Pharmacol52:243–251[CrossRef]
    [Google Scholar]
  43. Waditee R., Hibino T., Tanaka Y., Nakamura T., Incharoensakdi A., Takabe T. 2001; Halotolerant cyanobacterium Aphanothece halophytica contains an Na+/H+ antiporter, homologous to eukaryotic ones, with novel ion specificity affected by C-terminal tail. J Biol Chem276:36931–36938[CrossRef]
    [Google Scholar]
  44. Zingone A., Enevoldsen H. O. 2000; The diversity of harmful algal blooms: a challenge for science and management. Ocean Coastal Manag43:725–748[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.26350-0
Loading
/content/journal/micro/10.1099/mic.0.26350-0
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error