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Lymphocyte metallothionein mRNA responds to marginal zinc intake in human volunteers

Published online by Cambridge University Press:  09 March 2007

Adrian K. Allan ,
Gabrielle M. Hawksworth ,
Leslie R. Woodhouse ,
Barbara Sutherland ,
Janet C. King  and
John H. Beattie
Adrian K. Allan
Affiliation:
Nutrient-Gene Interaction Group, Rowett Research Institute, Aberdeen, UK
Gabrielle M. Hawksworth
Affiliation:
Departments of Medicine and Therapeutics and Biomedical Sciences, University of Aberdeen, Aberdeen, UK
Leslie R. Woodhouse
Affiliation:
Department of Nutritional Sciences, University of California at Berkeley, Berkeley, CA, USA
Barbara Sutherland
Affiliation:
Department of Nutritional Sciences, University of California at Berkeley, Berkeley, CA, USA
Janet C. King
Affiliation:
Department of Nutritional Sciences, University of California at Berkeley, Berkeley, CA, USA
John H. Beattie*
Affiliation:
Nutrient-Gene Interaction Group, Rowett Research Institute, Aberdeen, UK
*
*Corresponding author: Dr John H. Beattie, fax +44 1224 716629, email [email protected]
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Abstract

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Marginal Zn deficiency is thought to be prevalent in both developed and developing countries. However, the extent of Zn deficiency is not known, due to the lack of a reliable diagnostic indicator. Blood plasma and erythrocyte concentrations of metallothionein (MT) reflect Zn status, but measurement of MT is dependent on the availability of sensitive immunoassays. Our aim was to show whether measurement of T lymphocyte MT-2A mRNA, using a competitive reverse transcriptase (RT)–polymerase chain reaction (PCR) assay, could indicate Zn status in human subjects in a residential Zn-depletion study. In the study, the Zn intake of seven volunteers was maintained at 13·7 mg/d for 5 weeks (baseline) followed by 4·6 mg/d for 10 weeks (marginal intake) and then 13·7 mg/d (repletion) for 5 weeks. The quantitative assay was developed using standard techniques and concentrations of MT-2A mRNA were normalized by reference to β-actin mRNA which was also measured by competitive RT–PCR assay. An alternative method of measuring the PCR product using capillary electrophoresis with laser-induced fluorescence detection was also evaluated. There was considerable inter-individual variation in MT-2A mRNA concentration and the mean level at the end of the baseline period was 10·3 (SE 3·7) fg MT-2A mRNA/pg β-actin mRNA, which then decreased by 64 % during the low Zn intake period. After repletion, MT-2A mRNA returned to baseline concentrations. In contrast, plasma Zn was unchanged by marginal Zn intake or repletion. The effect of low Zn in all individuals was consistent. We conclude that this assay is a sensitive method of evaluating marginal changes in dietary Zn intake.

Keywords

MetallothioneinRT–PCRZinc deficiencyCapillary electrophoresis
Type
Research Article
Information
British Journal of Nutrition , Volume 84 , Issue 5 , November 2000 , pp. 747 - 756
Copyright
Copyright © The Nutrition Society 2000

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