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Stimulation of vascular protein synthesis by activation of oestrogen receptor beta
M Liang1, E Ekblad, J A Gustafsson
1Department of Physiological Sciences, Lund University, BMC F12, S-221 84 Lund, Sweden.
Abstract:
The objective of this study was to investigate the effects of oestrogen receptor (ER) beta activation on vascular protein synthesis and protein expression. Nuclear immunoreactivity towards ER beta was observed abundantly in vascular smooth muscle and endothelial cells of mouse aorta. No ER alpha-positive cell nuclei were observed. In aorta from ovariectomized mice, treatment with the selective ER beta agonist genistein (100 nM) for 24 h increased [(3)H]leucine incorporation by about 30%. This effect was prevented by the ER blocker ICI 182780 (10 microM). Although genistein treatment stimulated protein synthesis, it caused no change in total protein determined either by the Lowry method on tissue homogenate or by densitometric scanning of protein bands (10-220 kDa) separated by SDS-PAGE. Separation of [(35)S]methionine-labelled proteins by SDS-PAGE did not reveal the protein(s) stimulated by genistein. DNA synthesis was not affected by 100 nM genistein, suggesting that genistein-induced stimulation of protein synthesis is not part of a growth response. Protein expression, determined by SDS-PAGE, was similar in aorta from ER beta-knockout and wild-type mice, suggesting that expression of vascular proteins does not depend solely on a functional ER beta gene. We suggest that activation of vascular ER beta stimulates synthesis of proteins and that this response is not associated with vascular growth.
Insights
Activation of estrogen receptor beta (ER beta) in mouse aorta stimulates protein synthesis but not vascular growth. This ER beta-mediated protein synthesis is not linked to overall protein expression levels.
Area of Science:
- Vascular Biology
- Endocrinology
- Molecular Biology
Background:
- Estrogen receptor beta (ER beta) is present in vascular smooth muscle and endothelial cells.
- The role of ER beta in vascular protein synthesis and expression requires further investigation.
Purpose of the Study:
- To investigate the effects of ER beta activation on vascular protein synthesis and expression.
- To determine if ER beta activation influences vascular growth.
Main Methods:
- Immunoreactivity for ER beta in mouse aorta.
- Measurement of [(3)H]leucine incorporation to assess protein synthesis.
- SDS-PAGE to analyze protein expression and identify specific proteins.
- Assessment of DNA synthesis to evaluate growth response.
Main Results:
- ER beta was abundant in aortic cells, with no ER alpha observed.
- Genistein (selective ER beta agonist) increased protein synthesis by 30% in ovariectomized mice, an effect blocked by ICI 182780.
- Genistein did not alter total protein levels or specific protein expression patterns.
- Genistein did not affect DNA synthesis, indicating no growth response.
Conclusions:
- Activation of vascular ER beta stimulates protein synthesis.
- This ER beta-induced protein synthesis is independent of vascular growth.
- Vascular protein expression does not solely depend on a functional ER beta gene.