Production and analysis of Bac-TRAP mice that express fluorescent proteins driven by thrombomodulin or endothelial cell protein C receptor promoter

2014 Fiscal Year Final Research Report

• Project/Area Number: 25670454
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Hematology
• Research Institution: National Cardiovascular Center Research Institute
• Principal Investigator: MIYATA TOSHIYUKI 独立行政法人国立循環器病研究センター, 研究所, 部長 (90183970)
• Co-Investigator(Kenkyū-buntansha): BANNO Fumiaki 独立行政法人国立循環器病研究センター, 研究所, 研究員 (00373514) ; TASHIMA Yuko 大阪大学, 微生物病研究所, 助教 (10423104) ; MARUYAMA Keiko 独立行政法人国立循環器病研究センター, 研究所, 流動研究員 (30712624)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Keywords: 血管内皮細胞 / 遺伝子発現 / トロンボモジュリン / プロテインC受容体 / マウス / トランスクリプトーム

Outline of Final Research Achievements

Endothelia in the arterial and venous circulation are heterogeneous in nature. In order to understand their heterogeneity, we developed novel BAC transgenic mice with cDNA encoding fusion proteins of a fluorescent protein, EGFP or mCherry, and a ribosomal protein L10a under control of the endothelial cell-specific thrombomodulin (TM) or endothelial cell protein C receptor (EPCR) promoter. We compared the expression intensities of transcribing endothelial-specific markers before and after immunoprecipitation with anti-EGFP antibody or anti-mCherry antibody and found that those markers are increased merely 2-3 fold after immunoprecipitation. We further examined the immunofluorescence staining of the heart and lung of the transgenic mice using anti-EGFP antibody or anti-mCherry antibody but failed to detect the endothelia positive for the antibody staining. We concluded that novel BAC transgenic mice we established showed low expression of EGFP-L10a or mCherry-L10a in endothelia.

Free Research Field

医歯薬学