Abstract
The development of cytogenetic methods applied to cells and tissues of marine invertebrates has been hampered by (1) a lack of in vitro cell lines, (2) inadequate karyotypic information (partly as a result of too few workers chasing too many organisms), and (3) the failure of their chromosomes to band satisfactorily. Compared to mammalian cytogenetics, our knowledge of marine invertebrates lags behind by several decades. With the current concern about mutagens in the marine environment, and the recognition that the cells of marine species have sensitivities to DNA-damaging agents similar to those of higher organisms, there is a need for methods which can be used (a) in environmental monitoring and (b) to screen potentially harmful substances in the laboratory. In the absence of in vitro cell lines, embryos and larvae have been used to provide a supply of dividing cells for mutation studies, although the advent of molecular methods has now brought with it the means to detect DNA damage without any need for the cells to be in a dividing state. Moreover, the use of FISH (Fluorescence In Situ Hybridisation) now makes it possible to study numerical and structural chromosomal aberrations with far greater accuracy than was previously possible. A new marine genotoxicity assay is described, based on the embryos and larvae of a tube-dwelling polychaete worm (Pomatoceros lamarkii), suitable for both laboratory studies and field monitoring. This new Pomatoceros assay provides, at the same time, a useful model for studying the consequences of adult exposure on the offspring. A novel application of marine cytogenetic research is the study of the evolutionary adaptations of invertebrates living in naturally polluted extreme environments viz. deep sea hydrothermal vents, which are typified by high levels of toxic heavy metals and radionuclides, substances known to inflict damage to DNA. Given these new methodological and conceptual advances, it is predicted that our understanding of the role played by mutation in the marine environment, both in an evolutionary and toxicological context, will increase dramatically over the next decade.
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References
Accomando, R., A. Viarengo, R. Bordone, M. Taningher, L. Canesi & M. Orunesu, 1991. A rapid method for detecting DNA strand breaks in Mytilus galloprovincialis Lam. induced by genotoxic xenobiotic chemicals. Int. J. Biochem. 23: 227–229.
Ahnstrom, G. & K. Erixon, 1973. Radiation induced strand breakage in DNA from mammalian cells. Int. J. radiat. Biol. 23: 285–289.
Anderson, S. L., J. E. Hose & J. R. Knezovich, 1994. Genotoxic and developmental effects in sea urchins are sensitive indicators of effects of genotoxic chemicals. Envir. Toxicol. Chem. 13: 1033–1041.
Atienzar, F., R. Child, A. Evenden, A. N. Jha, D. Sawa, C. Walker & M. H. Depledge, 1998. Application of the arbitrarily primed polymerase chain reaction for the detection of DNA damage. Mar. envir. Res. 46: 331–335.
Bauman, J. G. J., J. Wiegant, P. Borst & P. Van Duijn, 1980. A new method for fluorescence microscopical localisation of specific DNA sequences by in situ hybridisation of fluorochrome-labeled RNA. Exp. Cell Res. 138: 485–490.
Bayne, B. L., 1976. Marine mussels: their ecology and physiology. Cambridge University Press, London.
Bihari, N., R. Batel & R. K. Zahn, 1992. Fractionation of DNA from marine invertebrate, (Mytilus galoprovincialis) hemolymph by alkaline elution. Comp. Biochem. Physiol. 102B: 419–424.
Boer, J. G., H. L. Erfle, D. Walsh, J. Holcroft & B. W. Glickman, 1996. The use of lad transgenic mice in genetic toxicology. In Pfeifer, G. F. (ed.), Technologies for Detection of DNA Damage and Mutations. Plenum Press, New York: 411–429.
Bolognesi, C., M. Panini, P. Roggieri, C. Ercolini & C. Pellegrino, 1992. Carcinogenic and mutagenic pollutants: impact on marine organisms. Map. Tech. Rep. Ser. 69: 113–121.
Burgeot, T., F. Galgani & E. His, 1995. The micronucleus assay in Crassostrea gigas for the detection of seawater genotoxicity. Mutat. Res. 342: 125–140.
Carter, S. B., 1967. Effects of cytochalasins on mammalian cells. Nature 213: 261–264.
Cornet, M., 1993. A short-term method for chromosome preparation from somatic tissues of adult mussel (Mytilus edulis). Experientia 49: 87–90.
Dixon, D. R., 1982. Aneuploidy in mussel embryos (Mytilus edulis L.) originating from a polluted dock. Mar. Biol. Letts. 3: 155161.
Dixon, D. R., 1985. Pomatoceros triqueter: a test system for environmental mutagenesis. In Bayne, B. L. (ed.), The Effects of Stress and Pollution on Marine Animals. Praeger Publishers, New York: 205–214.
Dixon, D. R. & K. R. Clarke, 1982. Sister chromatid exchange - a sensitive method for detecting damage caused by exposure to environmental mutagens in the chromosomes of adult Mytilus edulis. Mar. Biol. Letts. 3: 163–172.
Dixon, D. R., L. R. J. Dixon, P. L. Pascoe & J. T. Wilson, 1998b. Chromosome numbers and genome size in deep-sea hydrothermal vent organisms. InterRidge News 7: 20–22.
Dixon, D. R. & N. Flavell, 1986. A comparative study of the chromosomes of Mytilus edulis and Mytilus galloprovincialis. J. mar. biol. Ass. U.K. 66: 219–228.
Dixon, D. R. & I. R. B. McFadzen, 1987. Heterochromatin in the interphase nuclei of the common mussel Mytilus edulis L. J. exp. mar. Biol. Ecol. 112: 1–9.
Dixon, D. R. & R. L. Pascoe, 1994. Mussel eggs as indicators of mutagen exposure in coastal and estuarine marine environments. In Sutcliffe, D. W. (ed.), Water Quality and Stress Indicators in Marine and Freshwater Systems: Linking Levels of Organisation. Freshwater Biological Association, Cumbria: 124–137.
Dixon, D. R., P. L. Pascoe & L. R. J. Dixon, 1998a. Karyotypic differences between two species of Pomatoceros, P. triqueter and P larmarckii ( Polychaeta: Serpulidae). J. mar. biol. Ass. U.K. 78: 1–14.
Dixon, D. R. & D. Pollard, 1985. Embryo abnormalities in the periwinkle, Littorina saxatilis, as indicators of stress in polluted marine environments. Mar. Poll. Bull. 16: 29–33.
Dixon, D. R. & H. Prosser, 1986. An investigation of the genotoxic effects of an organotin antifouling compound (bis(trybutyltin)oxide) on the chromosomes of the edible mussel, Mytilus edulis. Aquat. Toxicol. 8: 185–195.
Dixon, D. R., J. T. Wilson, P. L. Pascoe & J. M. Parry, 1999. Anaphase aberrations in the embryos of the marine tubeworm Pomatoceros lamarckii (Polychaeta: Serpulidae): a new in vivo test assay for detecting aneugens and clastogens in the marine environment. Mutagenesis 14: 375–383.
Drouin, R. et al., 1996. Cupric ion/ascorbate/hydrogen peroxide-induced DNA damage: DNA-bound copper ion primarily induces hase modification. Free Radic. Biol. Med. 21: 261–273.
Everaarts, J. M., 1995. DNA integrity as a biomarker of marine pollution: strand breaks in seastar (Asterias rubens) and dab (Limanda limanda). Mar. Poll. Bull. 31: 431–438.
Fenech, M. and A. A. Morley, 1985. Cytokinesis-block micronucleus method in human lymphocytes: effect on aging and low-dose x-irradiation. Cytobios 43: 233–246.
Fenech, M. & S. Neville, 1992. Conversion of excision-repairable DNA lesions to micronuclei within one cell-cycle in human lymphocytes. Envir. Mol. Mutagen. 19: 27–36.
Gardner, G. R., R. J. Pruell & A. R. Malcolm, 1992. Chemical induction of tumors in oysters by a mixture of aromatic and chlorinated hydrocarbons, amines and metals. Mar. envir. Res. 34: 59–63.
Gossen, J. A., W. J. F. De Leeuw, C. H. T. Tan, E. C. Zwarhoff, F. Berends, P. H. M. Lohman, D. L. Knook and J. Vijg, 1989. Efficient rescue of integrated shuttle vectors from transgenic mice: A model for studying mutation in vivo. Proc. natnl. Acad. Sci. U.S.A. 86: 7971–7975.
Govan, H. L., Y. Vallesayoub & J. Braun, 1990. Fine mapping of DNA damage and repair in specific genomic segments. Nucl. Acids Res. 18: 3823–3830.
Gupta, R. C. & K. Randerath, 1988. Analysis of DNA adducts by 32P-postlabelling and Thin Layer Chromatography. In Friedberg, E. & P. H. Hanawalt (eds), DNA Repair. Marcel Dekker, New York: 399–418.
Hagmar, L. et al., 1994. Cancer risk in humans predicted by increased levels of chromosomal aberrations in lymphocytes–nordic study group on the health risk of chromosome damage. Cancer Res. 54: 2919–2922.
Harrison, F. L. & I. M. Jones, 1982. An in vivo sister chromatid exchange assay in the larvae of the mussel Mytilus edulis -response to three mutagens. Mutat. Res. 105: 235–242.
Harvey, J. S., B. P. Lyons, M. Waldock & J. M. Parry, 1997. The application of the 32P-postlabelling assay to aquatic biomonitoring. Mutat. Res. 378: 77–88.
Hassold, T. D., 1986. Chromosome abnormalities in human reproductive wastage. Trends Genet. 2: 105–110.
Heddle, J. A., 1973. A rapid in vivo test for chromosome damage. Mutat. Res. 18: 187–192.
Hook, E. B., 1983. Contribution of chromosome abnormalities to human morbidity and mortality and some comments upon surveillance of chromosome mutation rate. Mutat. Res. 114: 389–423.
Hose, J. E. & H. W. Puffer, 1983. Cytologic and cytogenetic anomalies induced in purple sea urchin embryos (Strongylocentrotus purpuratus S.) by parental exposure to benzo(a)pyrene. Mar. Biol. Letts. 4: 87–95.
Hose, J. E., H. W. Puffer, P. S. Oshida & S. M. Bay, 1983. Developmental and cytogenetic abnormalities induced in the purple sea urchin by environmental levels of benzo(a)pyrene. Arch. envir. Contam. Toxicol. 12: 319–325.
Jha, A. N., T. H. Hutchinson, J. M. Mackay, B. M. Elliott & D. R. Dixon, 1996. Development of an in vivo genotoxicity assay using the marine worm Platynereis dumerilii ( Polychaeta: Neridae). Mutat. Res. 359: 141–150.
John, H., M. Birnstiel & K. Jones, 1969. RNA-DNA hybrids at the cytological level. Nature 223: 582–587.
Jones, I. M. & F. L. Harrison, 1987. Variability in the frequency of sister-chromatid exchange in larvae of Mytilus edulis: implications for field monitoring. J. exp. mar. Biol. Ecol. 113: 283–288.
Klaude, M.. S. Eriksson, J. Nygren & G. Ahnstrom, 1996. The comet assay: mechanisms and technical considerations. Mutat. Res. 363: 89–96.
Kohler, S. W., G. S. Provost, A. Fieck, P. L. Kretz, W. O. Bullock, D. L. Putman, J. A. Sorge & J. M. Short, 1991. Analysis of spontaneous and induced mutations in transgenic mice using a lambda ZAPllacl shuttle vector. Envir. Mol. Mutagen. 18: 316–321.
Kohn, K. W., C. K. Erickson, A. G. Ewig & L. A. Zwelling, 1976. Fractionation of DNA from mammalian cells by alkaline elution. Biochemistry 15: 4629–4637.
Korenberg, J. R. and E. F. Freelander, 1974. Giemsa technique for the detection of sister chromatid exchanges. Chromosoma 48: 355. Li, G. and D. Hedgecock, 1998. Genetic heterogeneity, detected by
PCR-SSCP, among samples of larval pacific oysters (Crassostrea gigas) supports the hypothesis of large variance in reproductive success. Can. J. Fish. aquat. Sci. 55: 1025–1033.
Levan, G. & F. Mitelman, 1977. Chromosomes and the aetiology of cancer. In de la Chapelle, A. & M. Sorsa (eds), Chromosomes Today. Elsevier. Amsterdam: 363–371.
Martinez-Lage, A., A. Gonzalez-Tizon & J. Mendez, 1994. Characteristics of different chromatin types in Mytilus galloprovincialis L. after C-banding, fluorochromc and restriction-endonuclease treatments. Heredity 72: 242–249.
Mix, M. C., 1986. Cancerous diseases in aquatic animals and their association with environmental pollutants: a critical literature review. Mar. envir. Res. 20: 1–141.
Moore, M. N., D. M. Lowe, D. R. Livingstone & D. R. Dixon, 1986. Molecular and cellular indices of pollutant effects and their use in environmental impact assessment. Wat. Sci. Tech. 18: 223–232.
Nacci, D. E., S. Cayula and E. Jackim, 1996. Detection of DNA damage in individual cells from marine organisms using the single cell gel assay. Aquat. Toxicol. 35: 197–210.
Nacci, D. E. & S. Nelson, 1992. Application of the alkaline unwinding assay to detect DNA strand breaks in marine bivalves. Mar. envir. Res. 33: 38–100.
Olive, R. L. & J. R. Banath, 1993. Induction and rejoining of radiation-induced DNA single-strand breaks: `tail moment’ as a function of position in the cell cycle. Mutat. Res. 294: 275–283.
Olive, P. L., J. P. Banath and R. E. Durand, 1990. Heterogeneity in radiation-induced DNA damage and repair in tumour and normal cells measured using the `comet’ assay. Radiation Res. 122: 8694.
Olive, P. L., D. Wlodek & J. P. Banath, 1991. DNA double-strand breaks measured in individual cells subjected to gel electrophoresis. Cancer Res. 51: 4671–4676.
Ostling, O. & K. J. Johanson, 1984. Microelectrophoretic study of radiation-induced DNA damage in individual mammalian cells. Biochem. Res. Communicat. 123: 291–298.
Pardue, M. L. & J. G. Gall, 1969. Molecular hybridisation of radioactive DNA to the DNA of cytological preparations. Proc. natnl. Acad. Sci. U.S.A. 64: 600–604.
Pascoe, P. L., S. J. Patton, R. Critcher & D. R. Dixon, 1995. Robertsonian polymorphism in the marine gastropod Nucella lapillus: advances in karyology using rDNA loci and NORs. Chromosoma 104: 455–460.
Perry, P. and S. Wolff, 1974. New giemsa method for the differential staining of sister chromatids. Nature 251: 156.
Pesch, G. G., C. E. Pesch & A. R. Malcolm, 1981a. Neanthes arenaceodentata, a cytogenetic model for marine genetic toxicology. Envir. Mutagen. 3: 386–387.
Pesch, G. G., C. E. Pesch & A. R. Malcolm, 1981 h. Neanthes arenaceodentata, a cytogenetic model for marine genetic toxicology. Aquat. Toxicol. 1: 301–311.
Russo, A., G. Priante & A. M. Tommasi, 1996. PRINS localization of centromeres and telomeres in micronuclei indicates that in mouse splenocytes chromatid non-disjunction is a major mechanism of aneuploidy. Mutat. Res. 372: 173–180.
Savage, J. R. K., 1993. Update on target theory as applied to chromosome aberrations. Envir. Mol. Mutagen. 22: 198–207.
Schmid, W., 1977. The micronucleus test. In Kilbey, B. J., M. Legator, W. Nichols & C. Ramel (eds), Handbook of Mutagenicity Test Procedures. Elsevier, Amsterdam: 235–242.
Shugart, L. R. & C. W. Theodakaris, 1994. Environmental genotoxicity: probing the underlying mechanisms. Envir. Health Perspect. 102: 13–17.
Singh, N. P., M. T. McCoy, R. R. Tice & E. L. Schneider, 1988. A simple technique for quantitation of low levels of DNA damage in individual cells. Exp. Cell Res. 175: 184–191.
Sole, M., C. Porte, X. Biosca, C. L. Mitchelmore, J. K. Chipman, D. R. Livingstone and J. Albaiges, 1996. Effects of the `Aegean Sea’ oil spill on biotransformation enzymes, oxidative stress and DNA-Adducts in digestive gland of the mussel (Mytilus edulis L). Comp. Biochem. Physiol. 113B: 257–265.
Stein, J. E., W. L. Reichert, M. Nishimoto & U. Varanasi, 1990. Overview of studies on liver carcinogenesis in English Sole from Puget Sound — evidence for a xenobiotic chemical etiology. Sci. total Envir. 94: 51–69.
Steinert, S. A., 1996. Contribution of apoptosis to observed DNA damage in mussel cells. Mar. envir. Res. 42: 253–259.
Taylor, J. H., P. S. Woods & W. L. Hughes, 1957. The organisation and duplication of chromosomes as revealed by autoradiographic studies using tritium-labelled thymidine. Proc. natnl. Acad Sci. U.S.A. 43: 122–128.
Theodorakis, C. W., S. J. Dsurney & L. R. Shugart, 1994. Detection of genotoxic insult as DNA strand breaks in fish blood cells by agarose-gel electrophoresis. Envir. Toxicol. Chem. 13: 1023–1031.
Thorson, G., 1946. Reproduction and larval development of Danish marine bottom invertebrates with special reference to the planktonic larvae in the sound (Oresund). Medd. Komm. Haverundersog (ser. Plankton ) 4: 170–173.
Tsutsui, T., H. Maizumi, J. A. McLachlan & J. C. Barrett, 1983. Aneuploidy induction and cell transformation by diethylstilbestrol. Cancer Res. 43: 3814–3821.
Venier, P. & S. Canova, 1996. Formation of DNA adducts in the gill tissue of Mytilus galloprovincialis treated with benzoapyrene. Aquat. Toxicol. 34: 119–131
Viarengo, A., L. Canesi, M. Pertica & D. R. Livingstone, 1991. Seasonal variations in the antioxidant defence systems and lipid peroxidation of the digestive gland of mussels. Comp. Biochem. Physiol. 100B: 187–190.
Vijg, J. & G. R. Douglas, 1996. Bacteriophage lambda and plasmid lacZ transgenic mice for studying mutations in vivo. In Pfeifer, G. F. (ed.), Technologies for Detection of DNA Damage and Mutations. Plenum Press, New York: 391–410.
Vukmirovik, M., N. Bihari, R. K. Zahn, W. E. G. Muller & R. Batel, 1994. DNA damage in marine mussel Mytilus galloprovincialis as a biomarker of environmental contamination. Mar. Ecol. Prog. Ser. 109: 165–171.
Welsh, J., D. Ralph and M. McClelland, 1995. In Innis, M. A., D. H. Gelfand and J. J. Sninsky (eds), PCR Strategies. Academic Press, London: 249–276.
Wilson, J. T., P. L. Pascoe, J. M. Parry & D. R. Dixon, 1998. Evaluation of the comet assay as a method for the detection of DNA damage in the cells of a marine invertebrate, Mytilus edulis L ( Mollusca: Pelecypoda). Mutat. Res. 399: 87–95.
Wrisberg, M. N., C. M. Bilbo & H. Spliid, 1992. Induction of micronuclei in hemocytes of Mytilus edulis and statistical analysis. Ecotoxicol. envir. Safety 23: 191–205.
Zakour, H. R., M. L. Landolt & R. M. Kocan, 1984. Sister chromatid exchange analysis in cultured peripheral blood leukocytes of the coldwater marine fish. Mar. envir. Res. 14: 499–500.
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Dixon, D.R., Wilson, J.T. (2000). Genetics and marine pollution. In: Solé-Cava, A.M., Russo, C.A.M., Thorpe, J.P. (eds) Marine Genetics. Developments in Hydrobiology, vol 144. Springer, Dordrecht. https://doi.org/10.1007/978-94-017-2184-4_3
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