title: Gene expression analyses of mouse aortic endothelium in response to atherogenic stimuli.
abstract: OBJECTIVE
  Endothelial cells are central to the initiation of atherosclerosis, yet there has been limited success in studying their gene expression in the mouse aorta. To address this, we developed a method for determining the global transcriptional changes that occur in the mouse endothelium in response to atherogenic conditions and applied it to investigate inflammatory stimuli.
  
  
  RESULTS
  We characterized a method for the isolation of endothelial cell RNA with high purity directly from mouse aortas and adapted this method to allow for the treatment of aortas ex vivo before RNA collection. Expression array analysis was performed on endothelial cell RNA isolated from control and hyperlipidemic prelesion mouse aortas, and 797 differentially expressed genes were identified. We also examined the effect of additional atherogenic conditions on endothelial gene expression, including ex vivo treatment with inflammatory stimuli, acute hyperlipidemia, and age. Of the 14 most highly differentially expressed genes in endothelium from prelesion aortas, 8 were also perturbed significantly by ≥ 1 atherogenic conditions: 2610019E17Rik, Abca1, H2-Ab1, H2-D1, Pf4, Ppbp, Pvrl2, and Tnnt2.
  
  
  CONCLUSIONS
  We demonstrated that RNA can be isolated from mouse aortic endothelial cells after in vivo and ex vivo treatments of the murine vessel wall. We applied these methods to identify a group of genes, many of which have not been described previously as having a direct role in atherosclerosis, that were highly regulated by atherogenic stimuli and may play a role in early atherogenesis.
authors.affiliation: From the Departments of Microbiology, Immunology, and Molecular Genetics (A.E., A.J.L.), Pathology and Laboratory Medicine (J.B.), Medicine (A.J.L.), and Human Genetics (A.J.L.), University of California, Los Angeles; and Bristol-Myers Squibb, Applied Genomics, Princeton, NJ (N.S., P.K., W.-p.Y.).
authors.forename: Ayca
authors.initials: A
authors.lastname: Erbilgin
authors.forename: Nathan
authors.initials: N
authors.lastname: Siemers
authors.forename: Paul
authors.initials: P
authors.lastname: Kayne
authors.forename: Wen-pin
authors.initials: WP
authors.lastname: Yang
authors.forename: Judith
authors.initials: J
authors.lastname: Berliner
authors.forename: Aldons J
authors.initials: AJ
authors.lastname: Lusis
mesh_terms: D000818:Animals; D001011:Aorta / Q000166:cytology*; D050197:Atherosclerosis / Q000235:genetics* / Q000473:pathology*; D002469:Cell Separation / Q000379:methods; D042783:Endothelial Cells / Q000166:cytology / Q000502:physiology*; D015870:Gene Expression / Q000502:physiology; D006949:Hyperlipidemias / Q000235:genetics / Q000473:pathology; D051379:Mice; D012313:RNA / Q000302:isolation & purification / Q000378:metabolism; D059467:Transcriptome; D017539:Tunica Intima / Q000166:cytology
chemical_list: D012313:RNA
keywords: atherosclerosis; endothelium; genetics