Abstract
LLC-PK1 cells, an established cell line from pig kidney with proximal tubule properties, were cultivated in vitro at confluence on plastic dishes. They were then exposed (apical side) to inorganic cadmium (CdCl2, 5 μM) for periods ranging between 1 to 24 h. Analysis of the cell supernatant after homogenisation and ultracentrifugation indicated that Cd taken up in the first 3 h was bound to cytosolic high molecular weight proteins, but was redistributed to low molecular weight proteins at later stages. Induction of Cd-metallothionein (Cd-Mt) synthesis, as judged from Cd-Mt binding to a specific anti-Cd-Mt antibody and from the rate of35S-cys incorporation into a specific protein fraction, was apparent 3–6 h after the addition of Cd to the incubation medium.
References
Berlin M, Hammarstrom L, Maunsbach AG (1964) Microautoradiographic localization of water-soluble cadmium in mouse kidney. Acta Radiol 2: 345–352
Bradford M (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein. Anal Biochem 72: 248–254
Cempel M, Webb M (1976) The time-course of cadmium-thionein synthesis in the rat. Biochem Pharmacol 25: 2067–2071
Cherian MG (1985) Rat kidney epithelial cell culture for metal toxicity studies. In Vitro Cell Dev Biol 21: 505–508
Enger MD, Hildebrand CE, Seagrave J, Tobey RA (1986) Cellular resistance to cadmium. In: Foulkes EC (ed) Cadmium. Springer, Berlin, pp 363–369
Failla ML, Cousins RJ, Mascenik MJ (1979) Cadmium accumulation and metabolism by rat liver parenchymal cells in primary monolayer culture. Biochem Biophys Acta 583: 63–72
Felley-Bosco E, Diezi J (1989) Fate of cadmium in rat renal tubules: a micropuncture study. Toxicol Appl Pharmacol 98: 243–251
Frazier JM, Puglese J (1978) Dose dependence of cadmium kinetics in the rat liver following intravenous injection. Toxicol Appl Pharmacol 43: 461–474
Friberg L, Piscator M, Nordberg GF, Kjellström T (1974) Cadmium in the environment. CRC Press, Cleveland, Ohio
Gerson RJ, Shaikh ZA (1982) Uptake and binding of cadmium and mercury to metallothionein in rat hepatocyte primary cultures. Biochem J 208: 465–472
Gstraunthaler GJA (1988) Epithelial cells in tissue culture. Renal Physiol Biochem 11: 1–42
Hazen-Martin DJ, Sens DA, Blackburn JG, Sens MA (1989) Cadmium nephrotoxicity in human proximal tubule cell cultures. In Vitro Cell Dev Biol 25: 784–790
Jin T, Nordberg GF, Nordberg M (1986) Uptake of cadmium in isolated kidney cells — influence of binding form and in vivo pretreatment. J Appl Toxicol 6: 397–400
Koizumi S, Sone T, Kimura M (1985) Induction of metallothionein synthesis in cultured cells derived from rabbit kidney. J Cell Physiol 125: 223–228
Stacey NH, Klaassen CD (1980) Cadmium uptake by isolated hepatocytes. Toxicol Appl Pharmacol 55: 448–455
Tohyama C, Shaikh ZA (1981) Metallothionein in plasma and urine of cadmium exposed rats determined by a single antibody RIA. Fundam Appl Toxicol 1: 1–7
Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci 76: 4350–4354
Vander Mallie RJ, Garvey JS (1978) Production and study of antibody produced against rat cadmium thionein. Immunochemistry 15: 857–868
Webb M, Daniel M (1975) Induced synthesis of metallothionein by pig kidney cells in vitro in response to cadmium. Chem Biol Interact 10: 269–276
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Felley-Bosco, E., Diezi, J. Cadmium uptake and induction of metallothionein synthesis in a renal epithelial cell line (LLC-PK1). Arch Toxicol 65, 160–163 (1991). https://doi.org/10.1007/BF02034945
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02034945