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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 J.Beattie@rri.sari.ac.uk
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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

References

Aggett, PJ (1989) Severe zinc deficiency Zinc in Human Biology pp. 259–280. [Mills, CF]. London: Springer-Verlag.Google Scholar
Baer, MT, King, JC, Tamura, T, Margen, S, Bradfield, RB, Weston, WL and Daugherty, NA (1985) Nitrogen utilization, enzyme activity, glucose intolerance and leucocyte chemotaxis in human experimental zinc deficiency. American Journal of Clinical Nutrition 41, pp. 1220–1235.CrossRefGoogle Scholar
Branca, F; 1989 An ELISA technique for the measurement of metallothionein in man. An approach to the assessment of zinc statusMSc Thesis.Google Scholar
Bremner, I, (1987) Nutritional and physiological significance of metallothionein. In Metallothionein II, pp. 81–87. [Kägi, JHR and Kojima, Y]. Basel/Boston: Birkhauser Verlag.CrossRefGoogle Scholar
Bremner, I (1993) Metallothionein in the diagnosis of zinc deficiency. In Essential and Toxic Trace Elements in Human Health and Disease: An Update, pp. 33–45. [Prasad, AS]. New York: Wiley-Liss Inc.Google Scholar
Celi, FS, Zenilman, ME and Shuldiner, AR (1993) A rapid and versatile method to synthesise internal standards for competitive PCR. Nucleic Acids Research 21, pp. 1047.CrossRefGoogle ScholarPubMed
Chen, X-C, Yin, T-A, He, J-S, Ma, Q-Y, Han, Z-M and Li, L-X (1985) Low levels of zinc in hair and blood, pica, anorexia and poor growth in Chinese pre-school children. American Journal of Clinical Nutrition 42, pp. 694–700.CrossRefGoogle Scholar
Davies, NT, Carswell, AJP, Mills, CF (1985) The effect of variation of dietary calcium intake on the phytate–zinc interaction in rats. In Trace Elements in Man and Animals–5, pp. 456–457. [Mills, CF, Bremner, I and Chesters, JK]. Slough: Commonwealth Agricultural Bureau.Google Scholar
Forster, E (1994) An improved general method to generate internal standards for competitive PCR. Biotechniques 16, pp. 18–20.Google ScholarPubMed
Gilliland, G, Perrin, S, Blanchard, K and Bunn, HF (1990) Analysis of cytokine mRNA and DNA: detection and quantitation by competitive polymerase chain reaction. Proceedings of the National Academy of Sciences USA 87, pp. 2725–2729.CrossRefGoogle ScholarPubMed
Glasel, JA (1995) Validity of nucleic-acid purities monitored by 260 nm/280 nm absorbency ratios. Biotechniques 18, pp. 62–63.Google Scholar
Golden, MHN (1989) The diagnosis of zinc deficiency. In Zinc in Human Biology. pp. 323–333. [Mills, CF].CrossRefGoogle Scholar
Food and Nutrition Board, National Research Council (1989) Recommended Dietary Allowances. Washington, DC: National Academy Press.Google Scholar
Hambidge, KM, Hambidge, C, Jacobs, M and Baum, JD (1972) Low zinc levels in hair, anorexia, poor growth and hypogeusia in children. Pediatric Research 6, pp. 868–887.CrossRefGoogle ScholarPubMed
Hamer, DH (1986) Metallothionein. Annual Reviews of Biochemistry 55, pp. 913–951.CrossRefGoogle ScholarPubMed
Harley, CB, Menon, CR, Rachubinski, RA and Nieboer, E (1989) Metallothionein mRNA and protein induction by cadmium in peripheral-blood leukocytes. Biochemical Journal 262, pp. 873–879.CrossRefGoogle Scholar
Prasad, AS, Meftah, S, Abdallah, J, Kaplan, J, Brewer, GJ, Bach, JF and Dardenne, M (1988) Serum thymulin in human zinc deficiency. Journal of Clinical Investigation 82, pp. 1202–1210.CrossRefGoogle ScholarPubMed
Meftah, S, Prasad, AS, Lee, DY and Brewer, GJ (1991) Ecto 5′ nucleotidase (5′NT) as a sensitive indicator of human zinc deficiency. Journal of Laboratory and Clinical Medicine 118, pp. 309–316.Google ScholarPubMed
Miller, LV, Hambidge, KM, Naake, VL, Hong, Z, Westcott, JL and Fennessey, PV (1994) Size of the zinc pools that exchange rapidly with plasma zinc in humans: alternative techniques for measuring and relation to dietary zinc intake. Journal of Nutrition 124, pp. 268–276.CrossRefGoogle ScholarPubMed
Roberts, M, Geiger, W and German, JB (2000) The revolution in microanalytic chemistry: a macro-opportunity for clinical nutrition. American Journal of Clinical Nutrition 71, pp. 434–437.CrossRefGoogle ScholarPubMed
Sandström, B, Arvidsson, B, Cederblad, A and Bjorn-Rasmussen, E (1980) Zinc absorption from composite meals. I. the significance of wheat extraction rate on zinc, calcium and protein content in meals based on bread. American Journal of Clinical Nutrition 33, pp. 739–745.CrossRefGoogle ScholarPubMed
Sato, M, Mehra, R and Bremner, I (1984) Measurement of plasma metallothionein-I in the assessment of zinc-deficient and stressed rats. Journal of Nutrition 114, pp. 1683–1689.CrossRefGoogle ScholarPubMed
Smit-Vanderkooy, PD and Gibson, RS (1987) Food consumption patterns of Canadian preschool children in relation to zinc and growth status. American Journal of Clinical Nutrition 45, pp. 609–616.CrossRefGoogle ScholarPubMed
Solomons, NW (1979) On the assessment of zinc and copper nutriture in man. American Journal of Clinical Nutrition 32, pp. 856–871.CrossRefGoogle ScholarPubMed
Sullivan, VK, Burnett, FR and Cousins, RJ (1998) Metallothionein expression is increased in monocytes and erythrocytes of young men during zinc supplementation. Journal of Nutrition 128, pp. 707–713.CrossRefGoogle ScholarPubMed
Thomas, L; 1996 The effect of zinc deprivation on protein, energy and zinc metabolism in manPhD thesis.Google Scholar
Tong, D, Schnreeberger, C, Leodolter, S and Zeillinger, R (1997) Quantitative determination of gene expression by competitive reverse-transcription-polymerase chain reaction in degraded RNA samples. Analytical Biochemistry 251, pp. 173–177.CrossRefGoogle ScholarPubMed
Vallee, BL and Falchuk, KH (1993) The biochemical basis of zinc physiology. Physiological Reviews 73, pp. 79–117.CrossRefGoogle ScholarPubMed
Walravens, PA, Krebs, NF and Hambidge, KM (1983) Linear growth of low income preschool children receiving a zinc supplement. American Journal of Clinical Nutrition 38, pp. 195–201.CrossRefGoogle ScholarPubMed
Yurkow, EJ and Makhijani, PR (1998) Flow cytometric determination of metallothionein levels in human peripheral blood lymphocytes: utility in environmental exposure assessment. Journal of Toxicology and Environmental Health 54, pp. 445–457.Google ScholarPubMed