Comparative study on metallothionein induction in whole testicular tissue and isolated Leydig cells

Abel, Josef; Ruiter, N. de; Kühn‐Velten, W. Nikolaus

doi:10.1007/bf02307313

Cited by 25 publications

(16 citation statements)

References 34 publications

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“…However, after 6 h Cd post-injection the testes continues to show an increased level of MT-1 mRNA compared to that of the liver. The increment of MT-1 mRNA in testes due to Cd is also substantiated in other studies [McKenna et al 1996;Ren et al 2003;Abel et al 1991]. However, there are also reports that indicate a decrease and little or no difference of MT-1 mRNA levels in the whole testes of Cd-injected rats or mice [Shiraishi and Waalkes 1994;Zhou et al 1999].…”

Section: Resultssupporting

confidence: 70%

Expression of Metallothionein-1 (MT-1) mRNA in the Rat Testes and Liver After Cadmium Injection

Mukhopadhyay

Mitra

Nandi

et al. 2009

Systems Biology in Reproductive Medicine

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Metallothioneins (MTs) belong to the family of stress proteins that are present in the majority of living organisms. The MTs play an important task in detoxifying heavy metals. The mammalian scrotal testis is known to be susceptible to cadmium (Cd) exposure. The present work focuses on the MT-1 isoform and aims to ascertain and confirm previous findings to answer whether rodent testes indeed contain MT-1 mRNA, whether its level is increased with Cd injection in liver and testes, and lastly what is the relative difference in the expression of MT-1 mRNA in liver and testes both with and without Cd injection. Adult male Wistar rats weighing 270-290 g received a subcutaneous injection of 4.0 mmol Cd/kg and were sacrificed by cervical dislocation 6 h later. RNA was isolated from testes as well as the liver. There were 2 replicates per treatment for RNA analyses. MT-1 mRNA levels were determined by semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR) analysis and then assessed by densitometry scanning. The results of RT-PCR clearly demonstrated that the rodent testes express MT-1 mRNA. The densitometry data shows that the expression of MT-1 mRNA increases with Cd treatment in testes. The relative level of MT1-mRNA is greater in the control-liver than in the control-testes. However, upon Cd injection, the level of testes MT-1 mRNA increases 2.16 fold. These results suggest that the testes respond to Cd for at least 6 h post injection through a transcriptional mechanism.

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Section: Resultssupporting

confidence: 70%

Expression of Metallothionein-1 (MT-1) mRNA in the Rat Testes and Liver After Cadmium Injection

Mukhopadhyay

Mitra

Nandi

et al. 2009

Systems Biology in Reproductive Medicine

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“…The concomitant formation of disulfide bonds in the ␤-domain was demonstrated using 1 H and 113 Cd NMR spectroscopy (11). However, the use of Cd 7 -MT in these experiments may modify the cysteine reactivity, because Cd-MT is considered to be more stable than Zn-MT (10,29,30). The function of MT in the detoxification of heavy metals has been attributed to the tight metal binding in the ␣-domain, whereas the Fig.…”

Section: Discussionmentioning

confidence: 99%

“…The mechanism of the antioxidant action of MT remains elusive. It has been shown in vitro that MT directly interacts with reactive oxygen and nitrogen species to prevent oxidative and nitrosative damage to macromolecules such as lipids, proteins, and DNA (5)(6)(7)(8). However, this interaction has never been demonstrated in vivo and would unlikely be a major mechanism for the antioxidant action of MT in vivo because the reactivity of reactive oxygen and nitrogen species would only allow their reaction with molecules that have a close proximity to their generation sites.…”

mentioning

confidence: 99%

Metallothionein Disulfides Are Present in Metallothionein-overexpressing Transgenic Mouse Heart and Increase under Conditions of Oxidative Stress

Feng¹,

Benz

Cai

et al. 2006

Journal of Biological Chemistry

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Metallothionein (MT) releases zinc under oxidative stress conditions in cultured cells. The change in the MT molecule after zinc release in vivo is unknown although in vitro studies have identified MT disulfide bond formation. The present study was undertaken to test the hypothesis that MT disulfide bond formation occurs in vivo. A cardiac-specific MT-overexpressing transgenic mouse model was used. Mice were administered saline as a control or doxorubicin (20 mg/kg), which is an effective anticancer drug but with severe cardiac toxicity at least partially because of the generation of reactive oxygen species. A differential alkylation of cysteine residues in MT of the heart extracts was performed. Free and metal-bound cysteines were first trapped by N-ethylmaleimide and the disulfide bonds were reduced by dithiothreitol followed by alkylation with radiolabeled iodoacetamide. Analyses of the differentially alkylated MTs in the heart extract by high preformance liquid chromatography, SDS-PAGE, Western blot, and mass spectrometry revealed that disulfide bonds were present in MT in vivo under both physiological and oxidative stress conditions. More disulfide bonds were found in MT under the oxidative stress conditions. The MT disulfide bonds were likely intramolecular and both ␣-and ␤-domains were involved in the disulfide bond formation, although the ␣-domain appeared to be more easily oxidized than the ␤-domain. The results suggest that under physiological conditions, the formation of MT disulfide bonds is involved in the regulation of zinc homeostasis. Additional zinc release from MT under oxidative stress conditions is accompanied by more MT disulfide bond formation. Previous studies have shown that metallothionein (MT)2 functions in cardiac protection against oxidative damage (1-4). The mechanism of the antioxidant action of MT remains elusive. It has been shown in vitro that MT directly interacts with reactive oxygen and nitrogen species to prevent oxidative and nitrosative damage to macromolecules such as lipids, proteins, and DNA (5-8). However, this interaction has never been demonstrated in vivo and would unlikely be a major mechanism for the antioxidant action of MT in vivo because the reactivity of reactive oxygen and nitrogen species would only allow their reaction with molecules that have a close proximity to their generation sites. Alternatively, in vitro studies have demonstrated that the multiple cysteine residues of MT can be oxidized. The sulfur clusters that bind zinc in MT create an oxidoreductive environment for zinc at a redox potential so low that MT can be readily oxidized by mild cellular oxidants, such as glutathione disulfide, with the release of zinc (9). This MT oxidation and zinc release process would play a critical role in the cellular response to oxidative and nitrosative stress.The high content of cysteine residues enables MT to avidly bind zinc and other metals (10). The binding of divalent metals to MT occurs in two dumbbell-shaped domains, of which the N-terminal ␤-domain usually...

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“…By modulating Zn supply, MT proteins may play a central role in the regulation of cell growth and differentiation. MT expression in testes was not upregulated following exposure to cadmium (Durnam and Palmiter, 1981;Abel et al, 1991;McKenna et al, 1996), and therefore the existence of metal-binding proteins other than MT has been suggested.…”

Section: Introductionmentioning

confidence: 95%

A Novel Testis-Specific Metallothionein-like Protein,Tesmin,Is an Early Marker of Male Germ Cell Differentiation

Sugihara

Wadhwa²,

Kaul

et al. 1999

Genomics

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Comparative study on metallothionein induction in whole testicular tissue and isolated Leydig cells

Cited by 25 publications

References 34 publications

Expression of Metallothionein-1 (MT-1) mRNA in the Rat Testes and Liver After Cadmium Injection

Expression of Metallothionein-1 (MT-1) mRNA in the Rat Testes and Liver After Cadmium Injection

Metallothionein Disulfides Are Present in Metallothionein-overexpressing Transgenic Mouse Heart and Increase under Conditions of Oxidative Stress

A Novel Testis-Specific Metallothionein-like Protein,Tesmin,Is an Early Marker of Male Germ Cell Differentiation

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