Premium
Quantitative Proteomic Analysis of Endothelial Cells in G‐protein Coupled Estrogen Receptor ( Gper1 ) Knockout Rats
Author(s) -
Galla Sarah,
Waghulde Harshal,
Cheng Xi,
Chakraborty Saroj,
Mell Blair,
Basrur Venkatesha,
Joe Bina
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.884.4
Subject(s) - endocrinology , medicine , knockout mouse , estrogen receptor , estrogen , receptor , gene knockout , biology , chemistry , gene , biochemistry , cancer , breast cancer
G‐protein coupled estrogen receptor ( Gper1 ), a previously identified orphan receptor, now known to be a receptor for estrogen and aldosterone, has been localized to blood vessels, heart, and the central nervous system. Gper1 knockout models have been developed in mice and rats and have shown increased blood pressure as compared to controls. In addition, a Gper1 agonist, G1, caused decreased blood pressure when injected into mice. Women with a GPERP16L amino acid substitution in their GPER1 protein have increased BP. In order to study the effects of this receptor further, we created a novel CRISPR/Cas9 gene deletion model of the Gper1 gene on a Dahl Salt‐Sensitive (S) rat background. Hemodynamic and vascular phenotypic studies demonstrated that the Gper1 knockout rats had lower blood pressure and improved vascular function compared to the S rat. Microbiotal and metabolomics studies showed that Gper1 knockout rats had a significantly altered microbiotal composition and an increased circulating level of the short chain fatty acid, acetate. Since the improved vascular function was observed to be occurring in an endothelial dependent manner, we hypothesize that deletion of Gper1 in endothelial cells causes quantitative alterations in proteins that are directly impacted by the function of Gper1 . To test this hypothesis and explore mechanisms behind the observed BP lowering effect in Gper1 knockout rats, a quantitative proteomic analysis was conducted on endothelial cells isolated and cultured from the thoracic aorta of Gper1 knockout rats and wildtype S rats. Endothelial cell cultures were digested and quantitatively analyzed by Mass Spectrometry. A total of 101 statistically significant differentially expressed proteins were identified, of which 97 proteins were with higher expression in the Gper1 knockout rats. Data prioritizing three of the most significantly upregulated proteins, Fibrillin‐1, Ephrin B1, and Cd99, as potential mechanistic links downstream of the functionality of Gper1 in endothelial cells will be presented. Support or Funding Information Funding for this work to BJ from the NHLBI/NIH (HL020176) is gratefully acknowledged.