Hostname: page-component-78c5997874-s2hrs Total loading time: 0 Render date: 2024-11-14T23:23:36.207Z Has data issue: false hasContentIssue false
  • English
  • Français

Impact of the toxic dinoflagellate Alexandriumcatenella on Pacific oyster reproductive output: application of flow cytometryassays on spermatozoa

Published online by Cambridge University Press:  17 April 2013

Nelly Le Goïc ,
Hélène Hégaret ,
Caroline Fabioux ,
Philippe Miner ,
Marc Suquet ,
Christophe Lambert  and
Philippe Soudant
Nelly Le Goïc*
Affiliation:
Laboratoire des Sciences de l’Environnement Marin (LEMAR), IUEM-UBO, Technopôle Brest Iroise, Plouzané, France
Hélène Hégaret
Affiliation:
Laboratoire des Sciences de l’Environnement Marin (LEMAR), IUEM-UBO, Technopôle Brest Iroise, Plouzané, France
Caroline Fabioux
Affiliation:
Laboratoire des Sciences de l’Environnement Marin (LEMAR), IUEM-UBO, Technopôle Brest Iroise, Plouzané, France
Philippe Miner
Affiliation:
Laboratoire de Physiologie des Invertébrés (LPI), IFREMER, Technopôle Brest Iroise, Plouzané, France
Marc Suquet
Affiliation:
Laboratoire de Physiologie des Invertébrés (LPI), IFREMER, Technopôle Brest Iroise, Plouzané, France
Christophe Lambert
Affiliation:
Laboratoire des Sciences de l’Environnement Marin (LEMAR), IUEM-UBO, Technopôle Brest Iroise, Plouzané, France
Philippe Soudant
Affiliation:
Laboratoire des Sciences de l’Environnement Marin (LEMAR), IUEM-UBO, Technopôle Brest Iroise, Plouzané, France
*
a Corresponding author:[email protected]
Get access

Abstract

The toxic dinoflagellate Alexandrium catenella recurrently blooms on thecoasts of France and produces Paralytic Shellfish Toxins (PSTs) that accumulate inbivalves. These toxins can affect various physiological functions including reproduction.The present study aims to validate measurements of sperm viability, DNA content andmitochondrial membrane potential in Pacific oyster Crassostrea gigasusing flow cytometry coupled with fluorescent markers, and to use thesemeasurements to assess the cellular parameters of sperm from Pacific oysters exposed toA. catenella. These parameters may influence fertilization,embryogenesis and larval development in free-spawning shellfish. Sperm viability and DNAcontent estimation were assessed using SYBR-14, which only penetrates cells with intactmembranes. Cell mortality was measured with propidium iodide (PI), which penetrates cellswith membrane damage. Mitochondrial membrane potential, used as an estimate ofmitochondrial function, was measured using JC-1 dye, which selectively enters intomitochondria and reversibly changes colour from green to orange as the membrane potentialincreases. To assess the effect of toxic algae on oyster sperm, broodstock (ripe oysters)were fed toxic (A. catenella) or non toxic (Heterocapsatriquetra) dinoflagellates at 250 cell ml-1 for 9 days. After thisexposure period, mature oysters were stripped and cellular responses of sperm analysed.Average DNA staining, as measured by SYBR-14, appeared lower and more variable in gametesfrom A. catenella-exposed oysters than in those from control oysters fedH. triquetra. Additionally, mitochondrial membrane potential of spermfrom A. catenella-exposed oysters was significantly higher (1.5 fold)than that of sperm produced by oysters fed H. triquetra. Both theincrease of mitochondrial membrane potential and the modification of DNA structure can beexpected to impact spermatozoa ability to fertilize oocytes and could thus impact relatedreproductive processes.

Keywords

Harmful algal bloomSperm qualityCellular parametersFlow cytometryPacific oysterCrassostrea gigasAlexandrium catenella
Type
Research Article
Information
Aquatic Living Resources , Volume 26 , Issue 3 , 2013 , pp. 221 - 228
Copyright
© EDP Sciences, IFREMER, IRD 2013

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Adams, S.L., Hessian, P.A., Mladenov, P.V., 2003, Flow cytometric evaluation of mitochondrial function and membrane integrity of marine invertebrate sperm. Invert. Reprod. Dev. 44, 45–51. CrossRefGoogle Scholar
Akcha, F., Spagnol, C., Rouxel, J., 2012, Genotoxicity of diuron and glyphosate in oyster spermatozoa and embryos. Aquat. Toxicol. 106-107, 104–113. CrossRefGoogle Scholar
Anderson, D.M., Alpermann, T.J., Cembella, A.D., Collos, Y., Masseret, Et, Montresor, M., 2012, The globally distributed genus Alexandrium: Multifaceted roles in marine ecosystems and impacts on human health. Harmful Algae 14, 10–35. CrossRefGoogle ScholarPubMed
Basti, L., Go, J., Higuchi, K., Nagai, K., Segawa, S., 2011, Effects of the toxic dinoflagellate Heterocapsa circularisquama on larvae of the pearl oyster Pinctada fucata martensii (Dunker, 1873). J. Shellfish Res. 30, 177–186. CrossRefGoogle Scholar
Basti, L., Nagai, K., Tanaka, Y., Segawa, S., 2013, Sensitivity of gametes, fertilization, and embryo development of the Japanese pearl oyster, Pinctada fucuta martensii, to the harmful dinoflagellate, Heterocapsa circularisquama. Mar Biol. 160, 211–219. CrossRefGoogle Scholar
Bricelj, V.M., MacQuarrie, S.P., 2007, effects of brown tide (Aureococcus anophagefferens) on hard clam, Mercenaria mercenaria, larvae and implications for benthic recruitment. Mar Ecol. Prog. Ser. 333, 147–159. CrossRefGoogle Scholar
Collos, Y., Gagné, C., Laabir, M, Vaquer, A., Cecchi, P., Souchu, P., 2004, Nitrogenous nutrition of Alexandrium catenella (Dinophyceae) in cultures and in Thau lagoon, southern France. J. Phycol. 40, 96–103. CrossRefGoogle Scholar
Cordelli, E., Eleuteri, P., Leter, G., Rescia, M., Spanò, M., 2005, Flow cytometry applications in the evaluation of sperm quality: semen analysis, sperm function and DNA integrity. Contraception 72, 273–279. CrossRefGoogle ScholarPubMed
Dong, Q., Eudeline, B., Allen, Jr. S.K., Tiersch, T.R., 2002, Factors affecting sperm motility of tetraploid Pacific oysters. J. Shellfish Res. 21, 719–723. Google Scholar
Erard-Le Denn E., Morlaix M., Dao J.C., 1990, Effects of Gyrodinium cf. aureolum on Pecten maximus (post larvae, juveniles and adults). In: Graneli E., Sundstrom B, Edler L., Anderson D.M. (Eds.) Toxic Marine Phytoplankton, Elsevier 132–136.
Favret, P., Lynn, J.W., 2010, Flow-cytometric analyses of viability biomarkers in pesticide-exposed sperm of three aquatic invertebrates. Arch. Environ. Contam. Toxicol. 58, 973–84. CrossRefGoogle ScholarPubMed
Flajshans, M., Cosson, J., Rodina, M., Linhart, O., 2004, The application of image cytometry to viability assessment in dual fluorescence-stained fish spermatozoa. Cell Biol. Int. 28, 955–959. CrossRefGoogle Scholar
Franco, A., Kellner, K., Mathieu, M., Lelong, C., Goux, D., Heude-Brethelin, C., 2011, Male germ cells of the Pacific oyster Crassostrea gigas: flow cytometry analysis, cell sorting and molecular expression. Aquat. Living Resour. 24, 237–245. CrossRefGoogle Scholar
Garner, D.L., Johnson, L.A., Yue, S.T., Roth, B.L., Haugland, R.P., 1994, Dual DNA staining assessment of bovine sperm viability using SYBR-14 and propidium iodide. J. Androl. 15, 620–629. Google ScholarPubMed
Garner, D.L., Johnson, L.A., 1995, Viability assessment of mammalian sperm using SYBR-14 and propidium iodide. Biol. Reprod. 53, 276–284 CrossRefGoogle ScholarPubMed
Gillan, L., Evans, G., Maxwell, W.M.C., 2005, Flow cytometric evaluation of sperm parameters in relation to fertility potential. Theriogenology 63, 445–457. CrossRefGoogle ScholarPubMed
Graham, J.K., 2001, Assessment of sperm quality: a flow cytometric approach. Anim. Reprod. Sci. 68, 239–24. CrossRefGoogle ScholarPubMed
Granmo, A., Havenhand, J., Magnusson, K, Svane, I., 1988, Effects of the planktonic flagellate Chrysochromulina polylepis Manton et Park on fertilization and early development of the ascidian Ciona intestinalis (L.) and the blue mussel Mytilus edulis L. J. Exp. Mar. Biol. Ecol. 124, 65–71. CrossRefGoogle Scholar
Guillard, R.R.L. andHargraves, P.E., 1993, Stichochrysis immobilis is a diatom, not a chrysophyte. Phycologia 32, 234–236. CrossRefGoogle Scholar
Haberkorn, H., Lambert, C., Le Goïc, N., Moan, J., Suquet, M., Guéguen, M., Sunila, I., Soudant, P., 2010, Effects of Alexandrium minutum exposure on nutrition-related processes and reproductive output in oysters Crassostrea gigas. Harmful Algae 9, 427–439. CrossRefGoogle Scholar
Haberkorn, H., Tran, D., Massabuau, J-C., Ciret, P., Savar, V., Soudant, P., 2011, Relationship between valve activity, microalgae concentration in the water and toxin accumulation in the digestive gland of the Pacific oyster Crassostrea gigas exposed to Alexandrium minutum. Mar. Pollut. Bull. 62, 1191–1197. CrossRefGoogle ScholarPubMed
Hégaret, H., Brokordt, K.B., Gaymer, C.F., Lohrmann, K.B., Garcia, C., Varela, D., 2012, Effects of the toxic dinoflagellate Alexandrium catenella on histopathogical and escape responses of the Northern scallop Argopecten purpuratus. Harmful Algae 18, 74–83. CrossRefGoogle Scholar
Kasai, T., Ogawa, K., Mizuno, K., Nagai, S., Uchida, Y., Ohta, S., Fujie, M., Suzuki, K., Hirata, S, Hoshi, K., 2002, Relationship between sperm mitochondrial membrane potential, sperm motility, and fertility potential. Japan Asian J. Androl. 4, 97–103. Google ScholarPubMed
Leverone, J., Blake, N.J., Pierce, R.H., Shumway, S.E., 2006, Effects of the dinoflagellate Karenia brevis on larval development in three species of bivalve mollusk from Florida. Toxicon 48, 75–84. CrossRefGoogle ScholarPubMed
Lewis, C. andGalloway, T., 2009, Reproductive consequences of paternal genotoxin exposure in marine invertebrates. Environ. Sci. Technol. 43, 928–933. CrossRefGoogle Scholar
Lewis, C., Ford, A.T., 2012, Infertility in male aquatic invertebrates: a review. Aquat. Toxicol. 120-121, 79–89. CrossRefGoogle ScholarPubMed
Lilly, E.L., Kulis, D.M., Gentien, P., Anderson, D.M., 2002, Paralytic shellfish poisoning toxins in France linked to human-introduced strain of Alexandrium catenella from the western Pacific: evidence from DNA and toxin analysis. J. Plankton Res. 24, 443–452. CrossRefGoogle Scholar
Lu, X.Y. andWu, R.S.S., 2005, UV induces reactive oxygen species, damages sperm, and impairs fertilization in the sea urchin Anthocidaris crassispina. Mar. Biol. 148, 51–57. Google Scholar
Marchetti, C., Jouy, N., Leroy-Martin, B., Defossez, A., Formstecher, P., Marchetti, P., 2004, Comparison of four fluorochromes for the detection of the inner mitochondrial membrane potential in human spermatozoa and their correlation with sperm motility. Hum. Reprod. 19 2267–2276. CrossRefGoogle ScholarPubMed
Mukai, C., Okuno, M., 2004, Glycolysis plays a major role for adenosine triphosphate supplementation in mouse sperm flagellar movement. Biol. Reprod. 71, 540–547. CrossRefGoogle Scholar
Navarro, J.M., Contreras, A.M., 2010, An integrative response by Mytilus chilensis to the toxic dinoflagellate Alexandrium catenella. Mar. Biol., 157, 1967–1974. CrossRefGoogle Scholar
Nice, H.E., 2005, Sperm motility in the Pacific oyster (Crassostrea gigas) is affected by nonylphenol. Mar. Pollut. Bull. 50, 1668–1674. CrossRefGoogle ScholarPubMed
Ogier de Baulny, B., LeVern, Y., Kerboeuf, D., Maisse, G., 1997, Flow cytometric evaluation of mitochondrial activity and membrane integrity in fresh and cryopreserved rainbow trout (Onchorhynchus mykiss) spermatozoa. Cryobiology 34, 141–149. CrossRefGoogle Scholar
Paniagua-Chavez, C.G., Jenkins, J., Segovia, M., Tiersch, T.R., 2006, Assessment of gamete quality for the eastern oyster (Crassostrea virginica) by use of fluorescent dyes. Cryobiology 53, 128–138. CrossRefGoogle ScholarPubMed
Reers, M., Smiley, S.T., Mottola-Hartshorn, C., Chen, A., Lin, M., Chen, L.B., 1995, Mitochondrial membrane potential monitored by JC-1 dye. Methods Enzymol. 260, 406–414. CrossRefGoogle ScholarPubMed
REPHY Ifremer, espèces toxiques et toxines : Zones touchées par une interdiction de vente des coquillages 1992–2001. http://envlit.ifremer.fr/var/envlit/storage/documents/dossiers/toxines10ans/pdf/secteursfermes.pdf.
Silva, P.F.N., Gadella, B.M., 2006, Detection of damage in mammalian sperm cells. Theriogenology 65, 958–978. CrossRefGoogle ScholarPubMed
Summerson, H.C., Peterson, C.H., 1990, Recruitment failure of the Bay Scallop, Argopecten irradians concentricus, during the first red tide, Ptychodiscus brevis, outbreak recorded in North Carolina. Estuaries 13, 322–331. CrossRefGoogle Scholar
Tang, Y.Z., Gobler, C.J., 2009, Cochlodinium polykrikoides blooms and clonal isolates from the northwest Atlantic coast cause rapid mortality in larvae of multiple bivalve species. Mar. Biol. 156, 2601–2611. CrossRefGoogle Scholar
Thomas C.A., Garner D.L., DeJarnette J.M., Marshall C.E., 1998, Effect of cryopreservation of bovine sperm organelle function and viability as determined by flow cytometry. Biol. Reprod. 58.3.786.
Whedon, W.F., Kofoid, C.A., 1936, Dinoflagellata of the San Francisco region. I. On the skeletal morphology of two new species, Gonyaulax catanella and G. acatenella. Univ. Calif. Publs. Zool. 41, 25–34. Google Scholar