Cellular cadmium distribution in the common winkle,Littorina littorea (L.) determined by X-ray microprobe analysis and histochemistry

Abstract

Various tissues of common winkles, Littorina littorea (L.), experimentally exposed to cadmium (Cd) chloride were examined using light and electron microscopy and their elemental composition determined by X-ray microanalysis and histochemistry. Membrane granules in gill epithelial cells with paddle cilia contain carbonates, phosphates and sulphides associated with different cations in different types of granules. Traces of Cd have been found only in those granules containing sulphur and iron. Nephrocytes also contain small amounts of this metal in the cytoplasm of excretory cells. X-ray microanalysis reveals that concretions of basophilic cells are minor sites for Cd sequestration while BTAN-ASSG stain for unbound Cd indicates that most of the Cd is located within the lysosomes of digestive cells in association with proteins. Low amounts of the metal have been evidenced in the granules of epithelial mantle cells rich in sulphur. The results also indicate that hemocytes contain granules of calcium phosphate and iron sulphide. Cd is also associated to sulphur rather than to phosphate. These hemocytes may act as Cd carrier from gills to kidney and digestive gland. A hypothetical pathway for Cd accumulation and detoxification is suggested.

Cellular and subcellular distribution of metals in molluscs

“…Chassard-Bouchaud and Galle (1988) Chassard-Bouchaud et al (1989) lis, Dreissena polymorpha) (Lowe and Moore, 1979;Marigómez et al, 1986aMarigómez et al, , 1990aRecio et al, 1988;Ireland and Marigómez 1992;Giamberini et al, 1996a,b). In addition, autometallography (AMG), which visualizes free or loosely bound metal ions in tissues by transforming them into insoluble sulfides (Danscher, 1984), has been applied to localize Cd, Cu, Hg, Pb, and Zn at the light microscope in cells of marine and freshwater bivalves (Janssen and Ertelt-Janssen, 1983;Hemelraad and Herwig, 1988;Soto et al, 1996a,b;Soto and Marigómez, 1997a,b) as well as in cells of terrestrial slugs (Marigómez et al, , 1997.…”

“…Adams et al (1997) used energy dispersive X-ray microanalysis to detect Al, Ba, Ca, Cu, Fe, Mg, Mn, P, Pb, S, and Zn in mineralized granules of the freshwater mussel Hyridella depressa. Marigómez et al (1990a) applied EPMA to describe the pattern of Cd uptake, storage, transport, and excretion in the winkle Littorina littorea. Occasionally, EPMA has been used in combination with histochemical techniques.…”

Contribution of X-Ray Spectroscopy to Marine Ecotoxicology: Trace Metal Bioaccumulation and Detoxification in Marine Invertebrates

“…In gastropod mollusks, granules have been found in different tissues such as the digestive gland (Mason & Nott, 1981), gills (Marigomez et al, 1990;Mason et al, 1984;Simkiss & Mason, 1983), gonads (Quintana et al, 1987) and conjunctive tissue (Bouquegneau & Martoja, 1982;Martoja et al, 1980;Martoja et al, , 1985;). In crustaceans, these structures have been found in the digestive tract (Bernard & Lane, 1961;Guary & Négrel, 1981;Hopkin & Nott, 1979Vogt & Quinitio, 1994) and in the cuticular hypodermic parenchyme (Barka 2007;White, 1978 as cited in Mason & Jenkins, 1995).…”

Recent advances in the histochemical staining of heavy metals by means of chelating agents, with special reference to cadmium

“…Copper, lead, manganese, and zinc were also stained with high optical density Sumi et al, , 1982. BTAN was used for the histochemical demonstration of cadmium in midge, winkle, and earthworm (Sumi et al, 1984a;Gupta and Sundararaman, 1990;Marigómez et al, 1990;Prento, 1991). To examine the effects of alkyl or halogen substituents on staining sensitivity, thiazolylazo-(alkylamino)phenol and halo-and alkylthiazolylazo-phenol were prepared, but their sensitivities were not so high (Sumi et al, 1984b(Sumi et al, , 1986.…”