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Induction of Metallothionein in Rat Liver by Zinc Exposure: A Dose and Time Dependent Study

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Abstract

Metallothioneins (MTs) are low molecular weight ubiquitous metalloproteins with high cysteine (thiol) content. The intracellular concentration of zinc (Zn) is tightly regulated and MT plays a crucial role in it. The present study investigates the relationship between the Zn status (as a function of Zn concentration and time) in the rat liver and the occurrence of hepatic MT. For dose dependent study, four experimental groups, one control and three receiving different levels of metal supplementation, were chosen [Group 1 control and Group 2, Group 3, Group 4 receiving subcutaneous dose of 10, 50 and 100 mg of Zn/kg body weight (in the form of ZnSO4·7H2O), respectively]. For the time dependent expression of MT, again four experimental groups, i.e. Group 5 control and Group 6, Group 7, Group 8 receiving 50 mg of Zn/kg body weight (in the form of ZnSO4·7H2O) subcutaneously and sacrificed at different time intervals after last injection i.e. 6, 18, 48 h, respectively were chosen. Isolation of MT was done by using combination of gel filtration and ion exchange chromatography while characterization of MT fraction was carried in the wavelength range 200–400 nm. Expression of MT was studied by using Western blot analysis. The results revealed that the MT expression increases with increasing the dose of Zn administered and maximum at 18 h after last Zn injection. Accumulation of MT with increase dose would help in maintaining the intracellular Zn concentration by its sequestration which further reduces the possibility of undesirable binding of Zn to other proteins significantly and maintains Zn homeostasis. The maximum expression of MT at 18 h is indicative of its half life.

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Abbreviations

MT:

Metallothionein

LD:

Lethal dose

MTF-1:

Metal transcription factor-1

MRE:

Metal responsive element

DEAE:

Diethylaminoethyl

BSA:

Bovine serum albumin

MWCO:

Molecular weight cut-off

A280 :

Absorbance at 280 nm

A205 :

Absorbance at 205 nm

ESI-MS:

Electrospray ionization mass spectrometry

LMCT:

Ligand to metal charge transfer

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Acknowledgements

The authors are grateful to Dr. M. P. Bansal and Dr. B. P. Mohanty for sharing their immense knowledge with us. Dr. Renuka Ganger is gratefully acknowledged for the support rendered during western blot analysis. This work is funded by University Grants Commission (UGC) [Grant No: F. No. 37-317/2009 (SR)], New Delhi, India, and UGC Department of Atomic Energy (UGC-DAE) Consortium for Scientific Research (Grant No: UGC-DAE-CSR-KC/CRS/13/TE-05/0842), Kolkatta, India. Roobee Garla is thankful to UGC, New Delhi, for providing financial assistance in the form of Junior/Senior Research Fellowship.

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  1. Department of Biophysics, Panjab University, Chandigarh, 160014, India

    Roobee Garla, Preeti Kango, Navneet Kaur Gill & M. L. Garg

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  1. Roobee Garla
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Correspondence to Roobee Garla.

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Garla, R., Kango, P., Gill, N.K. et al. Induction of Metallothionein in Rat Liver by Zinc Exposure: A Dose and Time Dependent Study. Protein J 36, 433–442 (2017). https://doi.org/10.1007/s10930-017-9737-7

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