Biochemical Genetics

, Volume 24, Issue 7–8, pp 635–642 | Cite as

Intestinal metallothionein in lethal-milk mice with systemic zinc deficiency

  • Arthur GriderJr.
  • Lawrence C. Erway


Lethal-milk (C57BL/6J-lm) mice over 12 months of age exhibit clinical signs of systemic Zn deficiency. Such lm mice have increased concentrations of metallothionein (MT) in the intestinal mucosa. Various concentrations of Cd or Zn were added to the drinking water. MT was assayed using the Cd-saturation/hemolysate method and for sulfhydryl concentration (MT has 33% cysteine residues) with Ellman's reagent. As assayed by both methods, mucosa from untreated lm mice contained approximately twice as much MT as did the C57BL/6J-(+lm/+lm) (B6) controls. Treatment with 150 and 500 ppm Zn removed the genotypic differences observed for the untreated and Cd-treated mice. These results are consistent with the lm mutation affecting Zn metabolism through impaired MT metabolism as measured for the intestinal mucosa. These studies do not eliminate the possibility that the liver may also contribute.

Key words

zinc cadmium metallothionein lethal milk intestine Ellman's reagent Cd saturation/hemolysate 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Avdeef, A., Zelazowski, A. J., and Garvey, J. S. (1985). Cadmium binding by biological ligands, 3. Five- and seven-cadmium binding in metallothionein: A detailed thermodynamic study. Inorg. Chem. 241928.Google Scholar
  2. Birchmeier, W., and Christen, P. (1971). Chemical evidence for syncatalytic conformational changes in aspartate aminotransferase. FEBS Lett. 18209.Google Scholar
  3. Bonewitz, R. F., Jr., and Howell, R. R. (1981). Synthesis of a metallothionein-like protein in cultured human skin fibroblasts: Relation to abnormal copper distribution in Menkes' disease. J. Cell. Physiol. 106339.Google Scholar
  4. Brown, B. W., and Hollander, M. (1977). Statistics: A Biomedical Introduction John Wiley and Sons, New York, London, Sydney, Toronto.Google Scholar
  5. Buhler, R. H. O., and Kagi, J. H. R. (1974). Human hepatic metallothioneins. FEBS Lett. 39229.Google Scholar
  6. Chou, W. S., Savage, J. E., and O'Dell, B. L. (1969). Role of copper in biosynthesis of intramolecular cross-links in chick tendon collagen. J. Biol. Chem. 2445785.Google Scholar
  7. Dickie, M. M. (1969). Personal communication. Mouse News Lett. 4130.Google Scholar
  8. Erway, L. C., and Grider, A., Jr. (1984). Zinc metabolism in lethal-milk mice: otolith, lactation, and aging effects. J. Hered. 75480.Google Scholar
  9. Habeeb, A. F. S. A. (1973). A sensitive method for localization of disulfide containing peptides in column effluents. Anal. Biochem. 5660.Google Scholar
  10. Hundley, J. M. (1950). Achromotrichia due to copper deficiency. Proc. Soc. Exp. Biol. Med. 74531.Google Scholar
  11. McClain, P. E., Wiley, E. R., Beecher, G. R., Anthony, W. L., and Hsu, J. M. (1973). Influence of zinc deficiency on synthesis and cross-linking of rat skin collagen. Biochim. Biophys. Acta 304457.Google Scholar
  12. Ohtake, H., and Koga, M. (1979). Purification and characterization of zinc-binding protein from the liver of the partially hepatectomized rat. Biochem. J. 183683.Google Scholar
  13. Ohtake, H., Suyemitsu, T., and Koga, M. (1983). Sea urchin (Anthocidaris crassipina) egg zinc-binding protein. Biochem. J. 211109.Google Scholar
  14. Onosaka, S., and Cherian, M. G. (1981). The induced synthesis of metallothionein in various tissues of rat in response to metals. I. Effect of repeated injection of cadmium salts. Toxicology 2291.Google Scholar
  15. Overnell, J. (1982). Copper metabolism in crabs and metallothionein: In vivo effects of copper II on soluble hepatopancreas metal binding components in the crab Cancer Pagurus containing varying amounts of cadmium. Comp. Biochem. Biophysiol. 73B555.Google Scholar
  16. Piletz, J. E., and Ganschow, R. E. (1978). Zinc deficiency in murine milk underlies expression of the lethal milk (lm) mutation. Science 199181.Google Scholar
  17. Piletz, J. E., and Ganschow, R. E. (1979). Lethal milk mutation results in dietary deficiency in nursing mice. Am. J. Clin. Nutr. 31560.Google Scholar
  18. Piletz, J. E., and Herschman, H. R. (1982). Induction of metallothionein by zinc in lethal milk mutant mice. Biochem. Genet. 201221.Google Scholar
  19. Piletz, J. E., Andersen, R. D., Berry, W., and Herschman, H. R. (1983). Synthesis and degradation of hepatic metallothionein in mice differing in susceptibility to cadmium mortality. Biochem. Genet. 21561.Google Scholar
  20. Probst, G. S., Bousquet, W. F., and Miya, T. S. (1977). Kinetics of cadmium-induced hepatic and renal metallothionein synthesis in the mouse. Toxicol. Appl. Pharmacol. 3951.Google Scholar
  21. Richards, M. P., and Cousins, R. J. (1976). Metallothionein and its relationship to the metabolism of dietary zinc in rats. J. Nutr. 1061591.Google Scholar
  22. Robyt, J. F., Ackerman, R. J., and Chittenden, C. G. (1971). Reaction of protein disulfide groups with Ellman's reagent: A case study of a number of sulfhydryl and disulfide groups in Aspergillus oryzae α-amylase, papain, and lysozyme. Arch. Biochem. Biophys. 147262.Google Scholar
  23. Smith, K. T., Cousins, R. J., Silbon, B. L., and Failla, M. L. (1978). Zinc absorption and metabolism by isolated, vascularly perfused rat intestine. J. Nutr. 1081849.Google Scholar
  24. Tsunoo, H., Kino, K., Nakajima, H., Hata, A., Huang, I. Y., and Yoshida, A. (1978). Mouse liver metallothionein: Purification, molecular weight, amino acid composition, and metal content. J. Biol. Chem. 2534172.Google Scholar

Copyright information

© Plenum Publishing Corporation 1986

Authors and Affiliations

  • Arthur GriderJr.
    • 1
  • Lawrence C. Erway
    • 1
  1. 1.Department of Biological SciencesUniversity of CincinnatiCincinnati

Personalised recommendations