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Copper Uptake and Retention in Liver Parenchymal Cells Isolated from Nutritionally Copper-Deficient Rats

Interfaculty Reactor Institute, Delft University of Technology, Mekelweg 15, 2629 JB Delft, The Netherlands ^* TNO Institute for Experimental Gerontology, 2280 HV Rijswijk, The Netherlands

Copper uptake and retention were studied in primary cultures of liver parenchymal cells isolated from copper-deficient rats. Male Sprague-Dawley rats were fed a copper-deficient diet (< 1 mg Cu/kg) for 10 wk. Copper-deficient rats were characterized by low copper concentrations in plasma and liver, anemia, low plasma ceruloplasmin oxidase activity and increased ⁶⁴Cu whole-body retention. Freshly isolated liver parenchymal cells from copper-deficient rats showed a higher ⁶⁴Cu influx, which was associated with a higher apparent V_max of 45 ± 4 pmol Cu·mg protein^-1·min^-1 as compared with 30 ± 3 pmol Cu·mg protein^-1·min^-1 for cells isolated from copper-sufficient rats. No significant difference in the apparent K_m (30 µmol/L) was observed. Relative ⁶⁴Cu efflux from cells from copper-deficient rats was significantly smaller than the efflux from cells from copper-sufficient rats after prelabeling as determined by 2-h efflux experiments. Analysis of the medium after efflux from cells from copper-deficient rats showed elevated protein-associated ⁶⁴Cu, suggesting a higher incorporation of radioactive copper during metalloprotein synthesis. Effects of copper deficiency persist in primary cultures of parenchymal cells derived from copper-deficient rats, and short-term cultures of these cells offer a prospect for the study of cell biological aspects of the metabolic adaptation of the liver to copper deficiency.

KEY WORDS: • copper • liver parenchymal cells • hepatic copper transport • rats

¹ Present address: Department of Nutrition and Food Science, University of Arizona, Tucson, AZ 85721.

Manuscript received 23 May 1990. Revision accepted 21 December 1990.

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