title: Metallothionein (MT) 1/2 expression in MT 1/2 and MT 3 knock-out mice and Long-Evans Cinnamon (LEC) rats.
abstract: Metallothionein (MT) is known to be involved in various physiological roles and diseases. However, a standard method for MT measurement has not been established until recently. Therefore, we have developed an easy and specific enzyme-linked immunosorbent assays (ELISA) to determine MT-1 and MT-2. In order to evaluate the method we developed, MT-1/2 in liver, kidney and brain was determined in wild type (WT), MT-1/2 knockout (KO) and MT-3 KO mice, with and without Cd treatment. MT 1/2 in urine was determined in genetically disordered LEC rats (an animal model of Wilson disease). MT-1/2 concentrations in the liver, kidney and brain in MT-1/2 KO mice were significantly lower compared to those of WT and MT-3 KO mice. MT-1/2 concentrations in the livers of WT mice significantly increased with Cd administration, but not in MT-1/2 KO mice. Similar results were observed by immunohistochemical staining. To confirm the molecular weight (MW) of MT detected in organs by the ELISA, analysis with a Sephadex G-75 was performed. Two peaks of MT-1/2 (MW small and large) were detected in WT and MT-3 KO mice. The small MT peak was mostly depleted in MT 1/2-KO mice, while a large MT peak remained. A significant increase in MT-1/2 concentration was detected in the urine of LEC rats with age and especially at the hepatitis stage. In conclusion, MT-1/2 ELISA and immunohistochemical staining was highly correlated with MT-1/2 determination in experimental animal specimens and could be a robust analytical tool for physiological and toxicological studies.
authors.affiliation: Graduate School of Health Sciences, Gunma University, 3-39-22 Showa-machi, Maebashi, Gunma 371-8514, Japan. nkyoumi@health.gunma-u.ac.jp
authors.forename: Kyoumi
authors.initials: K
authors.lastname: Nakazato
authors.forename: Katsuyuki
authors.initials: K
authors.lastname: Nakajima
authors.forename: Takamitsu
authors.initials: T
authors.lastname: Nakano
authors.forename: Tsukasa
authors.initials: T
authors.lastname: Kodaira
authors.forename: Kenji
authors.initials: K
authors.lastname: Nakayama
authors.forename: Masahiko
authors.initials: M
authors.lastname: Satoh
authors.forename: Takeaki
authors.initials: T
authors.lastname: Nagamine
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism; D003300:Copper / Q000652:urine; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D007668:Kidney / Q000378:metabolism; D008099:Liver / Q000378:metabolism; D008297:Male; D008668:Metallothionein / Q000737:chemistry / Q000502:physiology* / Q000652:urine; D000090023:Metallothionein 3; D051379:Mice; D018345:Mice, Knockout; D008970:Molecular Weight; D051381:Rats; D020332:Rats, Inbred LEC; D015032:Zinc / Q000652:urine
chemical_list: D000090023:Metallothionein 3; C000715430:Mt3 protein, mouse; D003300:Copper; D008668:Metallothionein; D015032:Zinc