{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,4]],"date-time":"2026-10-04T12:21:12Z","timestamp":1791116472899,"version":"4.1.0"},"reference-count":33,"publisher":"American Association for the Advancement of Science (AAAS)","issue":"5535","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Science"],"published-print":{"date-parts":[[2001,8,31]]},"abstract":"<jats:p>\n            The coagulation protease thrombin triggers fibrin formation, platelet activation, and other cellular responses at sites of tissue injury. We report a role for PAR1, a protease-activated G protein\u2013coupled receptor for thrombin, in embryonic development. Approximately half of\n            <jats:italic>Par1<\/jats:italic>\n            <jats:sup>\u2013\/\u2013<\/jats:sup>\n            mouse embryos died at midgestation with bleeding from multiple sites. PAR1 is expressed in endothelial cells, and a PAR1 transgene driven by an endothelial-specific promoter prevented death of\n            <jats:italic>Par1<\/jats:italic>\n            <jats:sup>\u2013\/\u2013<\/jats:sup>\n            embryos. Our results suggest that the coagulation cascade and PAR1 modulate endothelial cell function in developing blood vessels and that thrombin's actions on endothelial cells\u2014rather than on platelets, mesenchymal cells, or fibrinogen\u2014contribute to vascular development and hemostasis in the mouse embryo.\n          <\/jats:p>","DOI":"10.1126\/science.1061259","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T09:53:24Z","timestamp":1027763604000},"page":"1666-1670","source":"Crossref","is-referenced-by-count":233,"title":["A Role for Thrombin Receptor Signaling in Endothelial Cells During Embryonic Development"],"prefix":"10.1126","volume":"293","author":[{"given":"Courtney T.","family":"Griffin","sequence":"first","affiliation":[{"name":"Cardiovascular Research Institute, University of California at San Francisco (UCSF), San Francisco, California 94143"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yoga","family":"Srinivasan","sequence":"additional","affiliation":[{"name":"Cardiovascular Research Institute, University of California at San Francisco (UCSF), San Francisco, California 94143"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yao-Wu","family":"Zheng","sequence":"additional","affiliation":[{"name":"Cardiovascular Research Institute, University of California at San Francisco (UCSF), San Francisco, California 94143"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei","family":"Huang","sequence":"additional","affiliation":[{"name":"Cardiovascular Research Institute, University of California at San Francisco (UCSF), San Francisco, California 94143"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shaun R.","family":"Coughlin","sequence":"additional","affiliation":[{"name":"Cardiovascular Research Institute, University of California at San Francisco (UCSF), San Francisco, California 94143"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"221","reference":[{"key":"e_1_3_1_2_2","unstructured":"R. W. Colman V. J. Marder E. W. Salzman J. Hirsh in Hemostasis and Thrombosis R. W. Colman V. J. Marder E. W. Salzman J. Hirsh Eds. (Lippincott Philadelphia 1994) pp. 3\u201318."},{"key":"e_1_3_1_3_2","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V95.4.1113.004k54_1113_1116"},{"key":"e_1_3_1_4_2","doi-asserted-by":"publisher","DOI":"10.1038\/35025229"},{"key":"e_1_3_1_5_2","doi-asserted-by":"publisher","DOI":"10.1038\/381516a0"},{"key":"e_1_3_1_6_2","doi-asserted-by":"crossref","unstructured":"T. H. Bugge et al. Proc. Natl. Acad. Sci. U.S.A. 93 6258 (1996).","DOI":"10.1073\/pnas.93.13.6258"},{"key":"e_1_3_1_7_2","doi-asserted-by":"crossref","unstructured":"P. Carmeliet et al. Nature 383 73 (1996).","DOI":"10.1038\/383073a0"},{"key":"e_1_3_1_8_2","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V88.5.1583.1583"},{"key":"e_1_3_1_9_2","unstructured":"J. Cui K. S. O'Shea"},{"key":"e_1_3_1_9_3","doi-asserted-by":"publisher","DOI":"10.1038\/384066a0"},{"key":"e_1_3_1_10_2","unstructured":"W. Y. Sun et al. Proc. Natl. Acad. Sci. U.S.A. 95 7597 (1998)."},{"key":"e_1_3_1_11_2","doi-asserted-by":"crossref","unstructured":"J. Xue et al. Proc. Natl. Acad. Sci. U.S.A. 95 7603 (1998).","DOI":"10.1073\/pnas.95.13.7603"},{"key":"e_1_3_1_12_2","unstructured":"A. L. Darrow et al. Thromb. Haemost. 76 860 (1996)."},{"key":"e_1_3_1_13_2","unstructured":"Mice and analyses. Par1 \u2013\/\u2013 (4) FV +\/\u2013 (8) and Fib +\/\u2013 (20) mice that had been backcrossed into the C57BL\/6J background (>97%) were used. TIE2p\/e- LacZ (19) transgenic mice were bred with Par1 \u2013\/\u2013 mice. To generate precisely timed pregnancies for Fig. 1 matings were restricted to a 1-hour interval (8 a.m. to 9 a.m.) with 9 a.m. counted as E0. In other cases matings were set up overnight and females were checked for vaginal plugs each morning before 10 a.m. Noon on the day of plug detection was counted as E0.5. Genotypes were assessed by Southern blot analysis of genomic DNA from embryonic or yolk sac tissue as described in supplementary material ( 28 ) . To analyze embryos for microscopic bleeding pregnant mice were killed and perfusion-fixed with 4% paraformaldehyde. To avoid artifacts related to dissection whole gravid uterine segments were then removed en bloc immersion-fixed overnight dehydrated embedded in paraffin sectioned (10 \u03bcm) and stained with hematoxylin and eosin. Whole-mount \u03b2-galactosidase staining and immunostaining for PECAM-1 (29) and in situ hybridization for PAR1 mRNA (17) and for an SV40 sequence tag in the TIE2p\/e-PAR1 transgene mRNA (30) were performed as described."},{"key":"e_1_3_1_14_2","doi-asserted-by":"publisher","DOI":"10.1038\/38284"},{"key":"e_1_3_1_15_2","unstructured":"G. R. Sambrano E. J. Weiss Y. W. Zheng W. Huang S. R. Coughlin Nature in press."},{"key":"e_1_3_1_16_2","doi-asserted-by":"crossref","unstructured":"R. A. Shivdasani et al. Cell 81 695 (1995).","DOI":"10.1016\/0092-8674(95)90531-6"},{"key":"e_1_3_1_17_2","unstructured":"R. Rugh The Mouse: Its Reproduction and Development (Oxford Univ. Press New York 1990)."},{"key":"e_1_3_1_18_2","unstructured":"S. J. Soifer K. G. Peters J. O'Keefe"},{"key":"e_1_3_1_18_3","first-page":"60","volume":"144","author":"Coughlin S. R.","year":"1994","unstructured":"Coughlin S. R., Am. J. Pathol. 144, 60 (1994).","journal-title":"Am. J. Pathol."},{"key":"e_1_3_1_19_2","unstructured":"Constructs. In the PAR1- LacZ knock-in targeting vector the start ATG and codons 2 to 4 of PAR1 exon 1 were replaced with a LacZ -PGK- Neo R cassette. A Kozak sequence immediately preceded the LacZ gene. A 1320\u2013base pair (bp) short arm 5\u2032 of the LacZ -PGK- Neo R casette a 4.5-kb long arm 3\u2032 of the LacZ -PGK- Neo R cassette and a 509-bp Southern probe 5\u2032 of the short arm (i.e. outside of the targeting construct) were generated by PCR of BAC DNA containing the PAR1 gene. Primers and Southern analysis were as described in supplementary material (28). The Neo R cassette was in the same transcriptional direction as PAR1 and was bounded by loxP sites. In vivo cre -mediated excision of the Neo R cassette using \u03b2-actin\u2013 cre transgenic mice decreased the intensity of \u03b2-galactosidase staining but did not change the overall pattern. For the TIE2p\/e-PAR1 transgenic construct a 1.2-kb Eco RI\/Xba I (blunted) fragment containing the PAR1 cDNA coding region was inserted 5\u2032 to an 847-bp fragment of SV40 small t intron and poly(A) sequence from the vector CPV2 (30) in pBluescript SK (Stratagene). A 2.1-kb Hind III fragment containing the TIE2 promoter and a 9-kb fragment containing the TIE2 enhancer from pHHNS (19) were inserted 5\u2032 and 3\u2032 to the 2-kb PAR1-SV40 respectively. The resulting 13.1-kb insert was used for microinjections."},{"key":"e_1_3_1_20_2","doi-asserted-by":"crossref","unstructured":"T. M. Schlaeger et al. Proc. Natl. Acad. Sci. U.S.A. 94 3058 (1997).","DOI":"10.1073\/pnas.94.7.3058"},{"key":"e_1_3_1_21_2","doi-asserted-by":"crossref","unstructured":"T. T. Suh et al. Genes Dev. 9 2020 (1995).","DOI":"10.1101\/gad.9.16.2020"},{"key":"e_1_3_1_22_2","unstructured":"J. H. Erlich et al. Am. J. Path. 157 1849 (2000)."},{"key":"e_1_3_1_23_2","doi-asserted-by":"crossref","unstructured":"M. J. Rabiet et al. Arterioscler. Thromb. Vasc. Biol. 16 488 (1996).","DOI":"10.1161\/01.ATV.16.3.488"},{"key":"e_1_3_1_24_2","doi-asserted-by":"publisher","DOI":"10.1091\/mbc.9.9.2639"},{"key":"e_1_3_1_25_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.274.34.23969"},{"key":"e_1_3_1_26_2","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.77.3.774"},{"key":"e_1_3_1_27_2","doi-asserted-by":"crossref","unstructured":"E. Papadimitriou et al. Am. J. Physiol. 272 C1112 (1997).","DOI":"10.1152\/ajpcell.1997.272.4.C1112"},{"key":"e_1_3_1_28_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)67551-9"},{"key":"e_1_3_1_29_2","unstructured":"Supplementary material is available on Science Online at www.sciencemag.org\/cgi\/content\/full\/293\/5535\/1666\/DC1"},{"key":"e_1_3_1_30_2","doi-asserted-by":"publisher","DOI":"10.1242\/dev.121.4.1089"},{"key":"e_1_3_1_31_2","doi-asserted-by":"publisher","DOI":"10.1172\/JCI1360"},{"key":"e_1_3_1_32_2","unstructured":"We thank D. Ginsburg of University of Michigan Ann Arbor for FV +\/\u2013 mice J. Degen of Children's Hospital Cincinnati for Fib +\/\u2013 mice T. Sato of University of Texas Southwestern for TIE2p\/e- LacZ transgenic mice and pHHNS DNA and G. Martin and B. Black of UCSF for \u03b2-actin\u2013 cre and smooth muscle \u03b1-actin promoter- LacZ transgenic mice respectively; R. Hamilton and J. Wong for the electron microscopy studies; L. Prentice and R. Advincula for technical assistance; G. Martin H. Bourne I. Herskowitz D. Stainier and P.-T. Chuang of UCSF for critical reading of this manuscript. C.T.G. was supported by a predoctoral fellowship from the American Heart Association Western States Affiliate. This work was supported in part by NIH HL44907 and HL65590."}],"container-title":["Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.science.org\/doi\/pdf\/10.1126\/science.1061259","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,1,9]],"date-time":"2024-01-09T22:12:13Z","timestamp":1704838333000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.science.org\/doi\/10.1126\/science.1061259"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2001,8,31]]},"references-count":33,"journal-issue":{"issue":"5535","published-print":{"date-parts":[[2001,8,31]]}},"alternative-id":["10.1126\/science.1061259"],"URL":"https:\/\/doi.org\/10.1126\/science.1061259","relation":{},"ISSN":["0036-8075","1095-9203"],"issn-type":[{"value":"0036-8075","type":"print"},{"value":"1095-9203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2001,8,31]]}}}