Abstract
In the present study, autometallography (AMG) has been performed on resin-embedded tissue sections, cryosections, as well as on isolated cells from the digestive gland, the gills and the haemolymph of mussels (Mytilus galloprovincialis) collected from the field. The AMG reaction was more intense in cryosections and isolated cells compared to resin sections. This intense AMG reaction found in cryosections and in isolated cells and the time effectiveness of the procedures enhances their use in a variety of studies, such as biomonitoring, even if small amount of metals are to be detected. In addition, the preparation of isolated cells does not require specific instruments or qualified personnel and, thus, the use of isolated cells is encouraging for AMG applications, although further laboratory investigation is required. On the other hand, the use of resin-embedded tissue sections for AMG applications, even though it is complicated and time consuming, leads to a high preservation of structural morphology and allows the exact localization of metals in discrete cellular compartments.
Similar content being viewed by others
References
Anagnostou C, Kamberi H, Karageorgis A (1998) Horizontal and vertical distribution of heavy metals in sediments from Thermaikos. Gulf Rapp Comm Int Mer Medit 35: 222.
Andersen JT, Baatrup E (1988) Ultrastructural localization of mercury accumulations in the gills, hepatopancreas, midgut and antennal glands of the brown schrimp, Cragon cragon. Aquat Toxicol 13: 309–324.
Baatrup E, Nielsen MG, Danscher G (1986) Histochemical demonstration of two mercury pools in trout tissues: Mercury in kidney and liver after mercuric chloride exposure. Ecotoxicol Environ Saf 12(3): 267–282.
Baron J (1991) In situ sites for xenobiotic activation and detoxification: Implications for the differential susceptibility of cells to the toxic actions of enviromental chemicals. Prog Histochem Cytochem 23: 32–44.
Cajaraville MP, Robledo Y, Etxeberria M, Marigomez I (1995) Cellular biomarkers as useful tools in the biological monitoring of environmental pollution: Molluscan digestive lysosomes. In: Cajaraville MP, ed. Cell Biology in Environmental Toxicology. Bilbo: University of the Basque Country Press Service, pp. 29–55.
Christensen M, Ellerman-Eriksen S, Rungby J, Mogensen SC, Danscher G (1991) Histochemical and functional evaluation of mercuric chloride toxicity in cultured macrophages. In: GraumannWand Drukker J, eds. Histo-and Cytochemistry as a Tool in Enviromental Toxicology. Progress in Histo-and Cytochemistry. Vol. 23. Stuttgart and New York: Fischer Verlag, pp. 306–315.
Christensen M, Rungby J, Mogensen SC (1989) Effects of selenium on toxicity and ultrastructural localization of mercury in cultured murine macrophages. Toxicol Lett 47(3): 259–270.
Dailianis S, Domouhtsidou GP, Raftopoulou E, Kaloyanni M, Dimitriadis VK (2003) Evaluation of neutral red retention assay, micronucleus test, acetylcholinesterase activity and a signal transduction molecule (cAMP) in tissues of Mytilus galloprovincialis (L.) in pollution monitoring. Mar Environ Res 56: 443–470.
Danscher G (1981) Histochemical demonstration of heavy metals. A revised version of the sulphide silver method suitable for both light and electron microscopy. Histochem 71: 1–16.
Danscher G (1984) Autometallography. A new technique for light and electron microscopic visualisation of metals in biological tissues (gold, silver, metal sulphides and metal selenides). Histochem 81: 331–335.
Danscher G (1991) Histochemical tracing of zinc, mercury, silver and gold. Prog Histochem Cytochem 23: 273–285.
Danscher G, Mller-Madsen B (1985) Silver amplification of mercury sulphides and selenides.Ahistochemical method for light and electron microscopic localization of mercury tissue. J Histochem Cytochem 31: 219–228.
Danscher G, Montagnese C (1994) Autometallographic localization of synaptic vesicular zinc and lysosomal gold, silver and mercury. J Histotechnol 17: 15–22.
Dimitriadis VK, Domouhtsidou GP, Raftopoulou E (2003) Localization of Hg and Pb in the palps, the digestive gland and the gills in Mytilus galloprovincialis (L.) using autometallography and x-ray microanalysis. Environ Pollut 125: 345–353.
Domouhtsidou GP, Dimitriadis VK (2000) Ultrastructural localization of heavy metals (Hg, Ag, Pb and Cu) in gills and digestive gland of mussels Mytilus galloprovincialis (L.). Arch Environ Contam Toxicol 38: 472–478.
Grelle C, Descamps M (2000) Advantages and limits of autometallography: Application to the earthworm Eisenia fetida typica. Bull Soc Zool De France 125(1): 9–16.
Hatzianestis I, Sklivagou E, Georgakopoulou E (2000) Organic contaminants in sediments and mussels from Thermaikos Gulf, Hellas. Toxicol Environ Chem 74: 203–216.
Lowe DM, Fossato VU, Depledge MH (1995) Contaminant induced lysosomal membrane damage in blood cells of mussels Mytilus galloprovincialis from the Venice Lagoon: An in vitro study. Mar Ecol Prog Ser 129: 189–196.
Marigomez IA, Angulo E, Moya J (1986) Cooper treatment of the digestive gland of the slug Arion ater L. 2. Morphometrics and histophysiology. Bull Environ Contam Toxicol 36: 608–615.
Marigomez I, Soto M, Cajaraville MP (1995) Morphofunctional patterns of cell and tissue systems involved in metal handling and metabolism. In: Cajaraville MP, ed. Cell Biology in Environmental Toxicology. Bilbo: University of the Basque Country Press Service, pp. 89–134.
Marigomez I, Soto M, Kortabitarte M (1996) Tissue — level biomarkers of biological effect of mercury on sentinel slugs Arion ater. Arch Environ Contam Toxicol 31: 54–62.
Marigomez I, Kortabitarte G, Dussart GBJ (1998) Tissue — level biomarkers in sentinel slugs as cost-effective tools to assess metal pollution in soils. Arch Environ Contam Toxicol 34: 167–176.
Marzabadi MR, Lovaas E (1996) Spermine prevent iron accumulation and depress lipofuscin accumulation in cultured myocardial cells. Free Radical Biol Med 21(3): 375–381.
Mason AZ, Nott JA (1980) A rapid, routine technique for the X-ray microanalysis of microincinerated cryosections: A SEM study of inorganic deposits in tissues of the marine gastropod Littorina littorea L. J Histochem Cytochem 28: 1301–1311.
Mersh J, Beauvais MN, Nagel P (1996) Induction of micronuclei in haemocytes and gill cells of zebra mussels, Dreissena polymorpha, exposed to clastogens. Mut Res 371: 47–55.
Morgan AJ (1985) X-ray microanalysis in electron microscopy for biologists. New York: Oxford University Press, pp. 79.
Owen G (1972) Lysosomes, peroxisomes and bivalves. Sci Prog Oxf 60: 299–318.
Reico A, Marigomez JA, Angulo E, Moya J (1988) Zinc treatment of the digestive gland of the slug Arion ater (L.) 1. Cellular distribution of zinc and calcium. Bull Environ Contam Toxicol 41: 858–864.
Rungby J, Ellermann-Eriksen S, Danscher G (1987) Effects of selenium on toxicity and ultrastructural localization of silver in cultured macrophages. Arch Toxicol 61(1): 40–45.
Soto M, Cajaraville MP, Marigomez I (1996) Tissue and cell distribution of copper, zinc and cadmium in the mussel, Mytilus galloprovincialis, determined by autometallography. Tissue Cell 28(5): 557–568.
Soto M, Kortabitarte M, Marigomez I (1995) Bioavailable heavy metals in estuarinewaters as assessed by metal/shell-weight indices in sentinel mussels Mytilus galloprovincialis. Mar Ecol Prog Ser 125: 127–136.
Soto M, Marigomez I (1997a) Metal bioavailability assessment in ‘mussel-watch’ programmes by automated image analysis of autometallographical black silver deposits (BSD) in digestive cell lysosomes. Mar Ecol Prog Ser 56: 141–150.
Soto M, Marigomez I (1997b) BSD extent, an index for metal pollution screening based on the metal content within digestive cell lysosomes of mussels as determined by autometallography. Ecotox Environ Saf 37: 141–151.
Soto M, Quincoces I, Marigomez I (1998) Autometallographical procedure for the localization of metal traces in molluscan tissues by light microscopy. J Histotechnol 21: 123–127.
Staffan PS, Sven Erik GN, Ulf TB (2001) Lipofuscin-formation in cultured retinal pigment epithelial cells is related to their melanin content. Free Radical Biol Med 30: 74–81.
Voutsinou-Taliadouri F, Hatzianestis I, Georgakopoulou-Gregoriadou E (1998) Trace elements, pesticides and PCBs levels in sediments of a bay influenced by anthropogenic activities (Thermaikos bay, N.W Aegean Sea). International Symposium on Marine Pollution, Extended synopses, Monaco, pp. 668–680.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Raftopoulou, E., Domouhtsidou, G. & Dimitriadis, V. Autometallographic Localization of Heavy Metals in Resin Sections, Cryosections and Isolated Cells of the Mussel Mytilus galloprovincialis (L.). Histochem J 34, 583–588 (2003). https://doi.org/10.1023/A:1026032714331
Issue Date:
DOI: https://doi.org/10.1023/A:1026032714331