Abstract
Majority of estrogen actions in the brain are mediated by estrogen receptor (ER) α which in turn is regulated by several factors like circulating levels of gonadal steroid hormones 17β-estradiol and testosterone, sex and age of the organism. The expression of ERα is regulated through interaction between cis-elements of its promoter and proteins present in the nuclei. Here, we have used electrophoretic mobility shift assay (EMSA) to analyze the effect of age, sex, 17β-estradiol, and testosterone on the binding of ERα promoter (−91 to +46 bp) to nuclear proteins from the mouse cerebral cortex. EMSA revealed the formation of three specific complexes in all groups. However, the intensity of these complexes varied as a function of age, sex and treatment with 17β-estradiol and testosterone. Nuclear proteins from the cerebral cortex of both sexes showed reduced binding with promoter fragment in old mice. Further, competition analysis indicated stronger binding in females than males of both ages. The extent of binding was reduced by 17β-estradiol and testosterone treatment in both ages and sexes. Thus, these findings demonstrate differential binding of nuclear proteins to mouse ERα promoter which may account for different functions of estrogen in the brain.
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Abbreviations
- DIG:
-
Digoxigenin
- DTT:
-
Dithiothreitol
- EDTA:
-
Ethylene diamine tetraacetic acid
- HEPES:
-
N-(2-hydroxyethyl) piperazine, N′-(2-ethane sulfonic acid)
- PMSF:
-
Phenyl methyl sulphonyl fluoride
- SDS:
-
Sodium dodecyl sulfate
- TBE:
-
Tris borate EDTA
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Acknowledgments
PKS acknowledges a research fellowship from the University Grants Commission, India. This work was supported by grants from the Department of Biotechnology, Government of India (BT/PR3593/Med/14/1468/2003) to MKT.
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Thakur, M.K., Sharma, P.K. Binding of estrogen receptor α promoter to nuclear proteins of mouse cerebral cortex: effect of age, sex, and gonadal steroids. Biogerontology 9, 467–478 (2008). https://doi.org/10.1007/s10522-008-9166-2
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DOI: https://doi.org/10.1007/s10522-008-9166-2