Selenomethionine stimulates expression of glutathione peroxidase 1 and 3 and growth of bovine mammary epithelial cells in primary culture
| Autor: | V.R. Osborne, N.G. Purdie, S.G. Miranda, Y. Wang, B.L. Coomber, John P. Cant |
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| Rok vydání: | 2009 |
| Předmět: |
GPX1
GPX3 Cell Survival Biology Gene Expression Regulation Enzymologic 03 medical and health sciences chemistry.chemical_compound Mammary Glands Animal Glutathione Peroxidase GPX1 Genetics medicine Animals Selenomethionine Cells Cultured Cell Proliferation 030304 developmental biology chemistry.chemical_classification Glutathione Peroxidase 0303 health sciences Glutathione peroxidase 030302 biochemistry & molecular biology Epithelial Cells Glutathione Immunohistochemistry Molecular biology Epithelium 3. Good health medicine.anatomical_structure Biochemistry chemistry Cell culture Dietary Supplements cardiovascular system biology.protein Female Animal Science and Zoology Immunostaining Food Science Peroxidase |
| Zdroj: | Journal of Dairy Science. 92:2670-2683 |
| ISSN: | 0022-0302 |
| DOI: | 10.3168/jds.2008-1901 |
| Popis: | This study examined the localization of cellular glutathione peroxidase (GPx1) and extracellular glutathione peroxidase (GPx3) in lactating mammary tissue and in primary cultures of bovine mammary epithelial cells (BMEC). The effect of selenium as selenomethionine (SeMet) on the growth and viability of BMEC and GPx protein expression and activity were also studied. Single mammary epithelial cells were recovered by serial collagenase/hyaluronidase digestion from lactating bovine mammary tissue and cultured in a low-serum collagen gel system enriched with lactogenic hormones and 0, 10, 20, or 50 nM SeMet. Positive immunostaining with anti-cytokeratin and bovine anti-casein confirmed the epithelial nature and differentiated state of BMEC. Addition of SeMet to media facilitated rapid confluence of BMEC and formation of dome structures. Immunohistochemical and immunocytochemical staining revealed that both GPx1 and GPx3 are synthesized by BMEC and localized in the cytoplasm and nucleus. Up to 50 nM SeMet linearly increased BMEC number and viability over 5 d of culture. Bovine mammary epithelial cells cultured in SeMet-supplemented medium also exhibited markedly elevated GPx activity and linear increases in abundance of GPx1 and GPx3 proteins. It is apparent that SeMet degradation to release Se for synthesis of selenoproteins is carried out by BMEC. Results indicate that bovine mammary epithelial cells express GPx1 and GPx3 in vivo and in vitro; SeMet enhances expression of these selenoproteins in vitro and the growth and viability of BMEC. |
| Databáze: | OpenAIRE |
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