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Toxicol Appl Pharmacol. 1995 Mar;131(1):144-54.

Constitutive expression of metallothionein genes in mouse brain.

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Department of Pharmacology, Toxicology and Therapeutics, University of Kansas Medical Center, Kansas City 66160.


Metallothioneins (MTs) are ubiquitous low-molecular-weight proteins that are induced by a variety of inducers, including metals, lipopolysaccharides (LPS), cytokines, oxidative stress, etc., and are thought to play a protective role against various toxic insults. The constitutive level of metallothionein is an important determinant of a tissue's susceptibility to toxic insults. In the present study, we report the constitutive expression of MT mRNAs in adult mouse brain. Analysis of total RNA from whole brain by Northern blot and solution hybridization showed that mRNAs for all three MT isoforms (I, II, III) were constitutively expressed in mouse brain, and there was no remarkable difference in their expression. However, in quantitative terms the order of expression was MT-I > MT-III > MT-II. The expression of MT-III and MT-II was about 70 and 50% of that of MT-I, respectively. Examination of their constitutive expression in different brain regions revealed that the three isoforms were expressed in all seven brain regions studied (olfactory bulb, cortex, caudate, hippocampus, thalamus, cerebellum, and brain stem), and there was only about a twofold difference in MT mRNA expression from one region of the brain to another. However, olfactory bulb had the highest mRNA expression for all three isoforms, as revealed by slot blot analysis. Constitutive expression of MT-I and -II mRNA, but not MT-III mRNA, was high in cerebellum. In order to study the cellular localization of MT mRNA, in situ hybridization of MT-I and MT-III mRNA was performed. For comparison, LPS was used to enhance MT-I mRNA signal because LPS is a good inducer of MT-I mRNA expression in mouse brain. In situ hybridization revealed that certain brain regions had distinctly localized high levels of expression of MT mRNAs. In brains of untreated mice, the constitutive expression of MT-I mRNA was high in the ependymal cell layer lining the lateral ventricles and in the Purkinje cell layer of cerebellum. The signal in the Purkinje cell layer was not on the Purkinje cells themselves, but was on locations consistent with that of glial cells. In LPS-treated mice, the signal in cerebellum was higher, and distinct signal appeared in the choroid plexus. However, signal in ependyma was similar to that in untreated mice. Pia mater in LPS-treated, but not in untreated, mouse brain showed enhanced signal for MT-I mRNA.(ABSTRACT TRUNCATED AT 400 WORDS).

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