Level of hydrogen peroxide affects expression and sub-cellular localization of Pax6

Original Article
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Abstract

The Pax6 is a multifunctional pairedbox and homeobox containing transcription factor which is involved in several functions of brain, eyes, and pancreas. It regulates expression of genes involved in cell proliferation, differentiation, inflammation, oxidative stress management, and neuropathy. Dynamic changes in the sub-cellular localization of Pax6 are proposed to regulate its activity, however, the underlying mechanism remains poorly understood. The oxidative stress mediated changes were studied in sub-cellular localization of Pax6 in cultured cells derived from the eye (cornea) and pancreas. The impact of induced oxidative stress was investigated on reactive oxygen species scavenger molecules, Superoxide dismutase1 (SOD1) and Catalase, and a critical cell signalling molecule Transforming growth factor-beta (TGF-β1). The cells were treated with three different concentrations of H2O2, viz., 0.3, 1.5, and 3.0 mM. The cell viability was analysed through Trypan blue dye exclusion assay. The localization of Pax6 was observed by immunofluorescence labeling, and alterations in levels of Pax6, SOD1, Catalase, and TGF-β1 were investigated by semi-quantitative RT-PCR. Nucleo-cytoplasmic shuttling of Pax6 was observed in cells of corneal epithelial (SIRC) and pancreatic origins (MIA-PaCa2). The percentage distribution of Pax6 in nuclear and cytoplasmic compartments of SIRC and MIA-PaCa2 cells was analyzed through ImageJ software. Level of hydrogen peroxide affects expression and sub-cellular localization of Pax6. Expression of Pax6 and TGF-β1 are directly associated with changes in sub-cellular localization of Pax6 and modulation in expression of Catalase. This may be the result of a cellular protective mechanism against peroxide-dependent cellular stress.

Keywords

Pax6 Oxidative stress Sub-cellular localization Nucleo-cytoplasmic shuttling 

Notes

Acknowledgements

Authors are highly thankful to Prof. M. K. Thakur, Biochemistry& Molecular Biology Unit, Department of Zoology, Banaras Hindu University, Varanasi, India, for providing the fluorescence microscope facility.

Compliance with ethical standards

Conflict of interest

Authors declare that no conflict of interest exist.

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Copyright information

© Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  1. 1.Biochemistry and Molecular Biology Laboratory, Department of Zoology, Institute of ScienceBanaras Hindu UniversityVaranasiIndia
  2. 2.Schepens Eye Research Institute, Massachusetts Eye and Ear InfirmaryHarvard Medical SchoolBostonUSA

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