{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,24]],"date-time":"2026-07-24T15:44:36Z","timestamp":1784907876444,"version":"3.55.0"},"reference-count":33,"publisher":"Wiley","issue":"19","license":[{"start":{"date-parts":[[2024,6,5]],"date-time":"2024-06-05T00:00:00Z","timestamp":1717545600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000274","name":"British Heart Foundation","doi-asserted-by":"publisher","award":["BHF, PG\/20\/1\/34617"],"award-info":[{"award-number":["BHF, PG\/20\/1\/34617"]}],"id":[{"id":"10.13039\/501100000274","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000274","name":"British Heart Foundation","doi-asserted-by":"publisher","award":["FS\/4yPhD\/F\/20\/34125"],"award-info":[{"award-number":["FS\/4yPhD\/F\/20\/34125"]}],"id":[{"id":"10.13039\/501100000274","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000266","name":"Engineering and Physical Sciences Research Council","doi-asserted-by":"publisher","award":["EP\/M012530\/1"],"award-info":[{"award-number":["EP\/M012530\/1"]}],"id":[{"id":"10.13039\/501100000266","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["bpspubs.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["British J Pharmacology"],"published-print":{"date-parts":[[2024,10]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background and Purpose<\/jats:title><jats:p>Thromboxane A<jats:sub>2<\/jats:sub> (TXA<jats:sub>2<\/jats:sub>) is a prostanoid produced during platelet activaton, important in enhancing platelet reactivity by activation of TP receptors. However, due to the short half\u2010life, studying TXA<jats:sub>2<\/jats:sub> signalling is challenging. To enhance our understanding of TP receptor\u2010mediated platelet biology, we therefore synthesised mono and difluorinated TXA<jats:sub>2<\/jats:sub> analogues and explored their pharmacology on heterologous and endogenously expressed TP receptor function.<\/jats:p><\/jats:sec><jats:sec><jats:title>Experimental Approach<\/jats:title><jats:p>Platelet functional and signalling responses were studied using aggregometry, Ca<jats:sup>2+<\/jats:sup> mobilisation experiments and immunoblotting and compared with an analogue of the TXA<jats:sub>2<\/jats:sub> precursor prostaglandin H<jats:sub>2<\/jats:sub>, U46619. G\u03b1<jats:sub>q<\/jats:sub>\/G\u03b1<jats:sub>s<\/jats:sub> receptor signalling was determined using a bioluminescence resonance energy transfer (BRET) assay in a cell line overexpression system.<\/jats:p><\/jats:sec><jats:sec><jats:title>Key Results<\/jats:title><jats:p>BRET studies revealed that F\u2010TXA<jats:sub>2<\/jats:sub> and F<jats:sub>2<\/jats:sub>\u2010TXA<jats:sub>2<\/jats:sub> promoted receptor\u2010stimulated TP receptor G\u2010protein activation similarly to U46619. Unexpectedly, F<jats:sub>2<\/jats:sub>\u2010TXA<jats:sub>2<\/jats:sub> caused reversible aggregation in platelets, whereas F\u2010TXA<jats:sub>2<\/jats:sub> and U46619 induced sustained aggregation. Blocking the IP receptor switched F<jats:sub>2<\/jats:sub>\u2010TXA<jats:sub>2<\/jats:sub>\u2010mediated reversible aggregation into sustained aggregation. Further BRET studies confirmed F<jats:sub>2<\/jats:sub>\u2010TXA<jats:sub>2<\/jats:sub>\u2010mediated IP receptor activation. F<jats:sub>2<\/jats:sub>\u2010TXA<jats:sub>2<\/jats:sub> rapidly and potently stimulated platelet TP receptor\u2010mediated protein kinase C\/P\u2010pleckstrin, whereas IP\u2010mediated protein kinase A\/P\u2010vasodilator\u2010stimulated phosphoprotein was more delayed.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion and Implications<\/jats:title><jats:p>F\u2010TXA<jats:sub>2<\/jats:sub> is a close analogue to TXA<jats:sub>2<\/jats:sub> used as a selective tool for TP receptor platelet activation. In contrast, F<jats:sub>2<\/jats:sub>\u2010TXA<jats:sub>2<\/jats:sub> acts on both TP and IP receptors differently over time, resulting in an initial wave of TP receptor\u2010mediated platelet aggregation followed by IP receptor\u2010induced reversibility of aggregation. This study reveals the potential difference in the temporal aspects of stimulatory and inhibitory pathways involved in platelet activation.<\/jats:p><\/jats:sec>","DOI":"10.1111\/bph.16435","type":"journal-article","created":{"date-parts":[[2024,6,6]],"date-time":"2024-06-06T03:20:11Z","timestamp":1717644011000},"page":"3685-3699","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Difluorinated thromboxane A<sub>2<\/sub> reveals crosstalk between platelet activatory and inhibitory pathways by targeting both the TP and IP receptors"],"prefix":"10.1111","volume":"181","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2327-5237","authenticated-orcid":false,"given":"Megan F.","family":"Allen","sequence":"first","affiliation":[{"name":"School of Physiology, Pharmacology and Neuroscience, Faculty of Life Sciences University of Bristol  Bristol 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