A mercury saturation assay for measuring metallothionein in fish

Journal Article · Thu Jul 01 04:00:00 EDT 1993 · Environmental Toxicology and Chemistry; (United States)

DOI:https://doi.org/10.1002/etc.5620120707· OSTI ID:5592413

Dutton, M D ^[1]; Stephenson, M ^[2]; Klaverkamp, J F ^[3]

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1. Univ. of Manitoba, Winnipeg (Canada). Dept. of Zoology
2. AECL Research, Pinawa, Manitoba (Canada). Environmental Science Branch
3. Freshwater Inst., Winnipeg, Manitoba (Canada)

An accurate, rapid, sensitive, and simple method using mercury saturation for quantifying metallothionein (MT) is described. A complex solution of enzymatic and nonenzymatic thiols, including rabbit liver MT-2, and supernatants from homogenized samples of rainbow trout liver were incubated in the presence of [sup 203]Hg in 10% trichloroacetic acid. Excess Hg was bound to an removed by chicken egg albumin, which denatured on contact with the acidic assay medium. After centrifugation, MT labeled with [sup 203]Hg remained in the TCA supernatant and was estimated using known stoichiometry for Hg-MT binding. A dilution series was used to establish that nonspecific metal binding, a common problem with other metal saturation assays, is negligible. Analysis of hepatic MT with high Cu content from rainbow trout demonstrated virtually complete displacement of Cu, Cd, and Zn by Hg. When compared to other metal-saturation assays developed for vertebrates, this method requires the least number of technical steps, and one-third or less of total preparatory and analytical time.

• OSTI ID: 5592413
• Journal Information: Environmental Toxicology and Chemistry; (United States), Journal Name: Environmental Toxicology and Chemistry; (United States) Vol. 12:7; ISSN 0730-7268; ISSN ETOCDK
• Country of Publication: United States
• Language: English

Related Subjects

560300* -- Chemicals Metabolism & Toxicology
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ANIMALS
AQUATIC ORGANISMS
BIOASSAY
BIOSYNTHESIS
CADMIUM
COPPER
ELEMENTS
FISHES
MERCURY
METABOLISM
METALLOPROTEINS
METALLOTHIONEIN
METALS
ORGANIC COMPOUNDS
PROTEINS
SENSITIVITY
SYNTHESIS
TRANSITION ELEMENTS
TROUT
VERTEBRATES
ZINC