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Metallothionein Gene Expression in Rats: Tissue-Specific Regulation by Dietary Copper and Zinc¹

Food Science and Human Nutrition Department, University of Florida, Gainesville, FL 32611

Regulation of metallothionein gene expression by dietary zinc and copper was examined in rat liver, kidney, intestine and brain using a 3 x 3 factorial design. Purified diets containing 5, 30 and 180 mg Zn/kg and 1, 6 and 36 mg Cu/kg were fed for 2 wk. Serum concentrations of copper and zinc were lower at the lowest intakes of either metal than at normal or supplemental levels. Kidney metallothionein levels were proportional to dietary zinc, being 50% less in the 5 mg Zn/kg group than in those fed the highest zinc intake. Metallothionein mRNA was measured by dot blot hybridization to a ³²P-labeled oligonucleotide DNA probe representing the terminal 5' sequence of the metallothionein gene. In kidney the number of metallothionein mRNA molecules per cell increased four- to five-fold (from 4 to 29 molecules per cell) with increasing dietary zinc. A less pronounced effect on metallothionein mRNA was observed in response to dietary copper. At the lowest copper intake level and highest intake of zinc intestinal metallothionein mRNA was sevenfold greater than in any other group. Liver and brain did not respond appreciably to the dietary levels of copper and zinc that were fed. Chromatography showed that copper and zinc content of renal metallothionein was directly related to the dietary levels fed. In kidney, both metallothionein-1 and -2 genes were expressed.

KEY WORDS: • metallothionein • zinc • gene expression • regulation • copper

¹ This research was supported by National Institutes of Health Grant No. DK 31127 from the National Institute of Diabetes, Digestive and Kidney Diseases and Boston Family Endowment Funds.

² To whom reprint requests and correspondence should be addressed.

Manuscript received 18 August 1987. Revision accepted 29 October 1987.