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Differential regulation of gene expression by ovariectomy in mouse aorta
Amparo C Villablanca1, Kristine A Lewis, Doris Tham
1Division of Cardiovascular Medicine, University of California, Davis, California 95616-8636, USA. avillablanca@ucdavis.edu
Abstract:
The purpose of this study was to investigate the effects of ovarian hormones on gene expression in the vascular wall. Our approach employed an RT-PCR-based cloning strategy of DNA differential display analysis and verification/confirmation of differential expression by semi-quantitative PCR and real-time PCR. mRNA analysis of normal aortas from intact and ovariectomized female C57BL/6J mice, showed altered expression of 20 genes with significant (>70%) sequence homology to known genes. Eight were selected for further study based on the genes' known function and potential relevance to vascular physiology. Differential expression of mRNA for three genes was confirmed by both semi-quantitative and real-time RT-PCR using gene-specific primers. Ovariectomy downregulated expression of elongation factor-1alpha (3.5-fold), ganglioside-induced differentiation associated protein (8.2-fold), and NADH:ubiquinone oxidoreductase (3.8-fold). Thus, in normal mouse aortas, ovariectomy resulted in significant differential downregulation of a number of vascular genes important to vascular cell growth and angiogenesis, cellular differentiation, and mitochondrial energy metabolism, respectively. These studies have implications for our understanding of hormonal regulation of vascular gene expression and the therapeutic targeting of specific vascular genetic sequences by female sex steroid hormones.
Insights
Ovarian hormones significantly impact vascular gene expression. Ovariectomy in mice downregulated key genes involved in vascular growth, differentiation, and metabolism, suggesting hormonal regulation of vascular health.
Area of Science:
- Vascular Biology
- Endocrinology
- Molecular Biology
Background:
- Ovarian hormones play a critical role in cardiovascular health.
- Understanding the molecular mechanisms of hormonal regulation in the vascular system is essential.
Purpose of the Study:
- To investigate the impact of ovarian hormones on gene expression within the vascular wall.
- To identify specific genes regulated by ovarian hormones in mouse aortas.
Main Methods:
- Utilized DNA differential display analysis with RT-PCR for gene cloning.
- Confirmed differential gene expression using semi-quantitative and real-time PCR.
- Analyzed mRNA from intact and ovariectomized mouse aortas.
Main Results:
- Identified 20 differentially expressed genes in mouse aortas post-ovariectomy.
- Confirmed downregulation of three key genes: elongation factor-1alpha, ganglioside-induced differentiation associated protein, and NADH:ubiquinone oxidoreductase.
- Observed significant fold changes in gene expression (3.5-8.2 fold) due to ovariectomy.
Conclusions:
- Ovariectomy leads to significant downregulation of vascular genes involved in cell growth, angiogenesis, differentiation, and energy metabolism.
- These findings highlight the importance of ovarian hormones in regulating vascular gene expression.
- Suggests potential for therapeutic targeting of vascular genetic sequences by female sex steroid hormones.