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Passage through the \u03b2-selection checkpoint requires the nascent TCR\u03b2 protein to mediate signaling through a pre-TCR complex. In this study, we show that developing T cells at the \u03b2-selection checkpoint establish an immunological synapse in in vitro and in situ, resembling that of the mature T cell. The immunological synapse is dependent on two key signaling pathways known to be critical for the transition beyond the \u03b2-selection checkpoint, Notch and CXCR4 signaling. In vitro and in situ analyses indicate that the immunological synapse promotes passage through the \u03b2-selection checkpoint. Collectively, these data indicate that developing T cells regulate pre-TCR signaling through the formation of an immunological synapse. This signaling platform integrates cues from Notch, CXCR4, and MHC on the thymic stromal cell to allow transition beyond the \u03b2-selection checkpoint.<\/jats:p>","DOI":"10.1083\/jcb.201908108","type":"journal-article","created":{"date-parts":[[2020,12,11]],"date-time":"2020-12-11T02:24:52Z","timestamp":1607653492000},"update-policy":"https:\/\/doi.org\/10.1083\/jcb.crossmarkpolicy","source":"Crossref","is-referenced-by-count":32,"title":["Developing T cells form an immunological synapse for passage through the \u03b2-selection checkpoint"],"prefix":"10.1083","volume":"220","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8743-3639","authenticated-orcid":true,"given":"Amr H.","family":"Allam","sequence":"first","affiliation":[{"name":"Optical Sciences Centre, Faculty of Science, Engineering & Technology, Swinburne University of Technology, Hawthorn, Victoria, Australia 1"},{"name":"Immune Signalling Laboratory, Peter MacCallum Cancer Centre, Parkville, Victoria, Australia 2"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3845-9290","authenticated-orcid":true,"given":"Mirren","family":"Charnley","sequence":"additional","affiliation":[{"name":"Optical Sciences Centre, Faculty of Science, Engineering & Technology, Swinburne University of Technology, Hawthorn, Victoria, Australia 1"},{"name":"Immune Signalling Laboratory, Peter MacCallum Cancer Centre, Parkville, Victoria, Australia 2"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kim","family":"Pham","sequence":"additional","affiliation":[{"name":"Optical Sciences Centre, Faculty of Science, Engineering & Technology, Swinburne University of Technology, Hawthorn, Victoria, Australia 1"},{"name":"Immune Signalling Laboratory, Peter MacCallum Cancer Centre, Parkville, Victoria, Australia 2"},{"name":"Department of Pathology, The University of Melbourne, Australia 3"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5826-9641","authenticated-orcid":true,"given":"Sarah M.","family":"Russell","sequence":"additional","affiliation":[{"name":"Optical Sciences Centre, Faculty of Science, Engineering & Technology, Swinburne University of Technology, Hawthorn, Victoria, Australia 1"},{"name":"Immune Signalling Laboratory, Peter MacCallum Cancer Centre, Parkville, Victoria, Australia 2"},{"name":"Department of Pathology, The University of Melbourne, Australia 3"},{"name":"Sir Peter MacCallum Department of Oncology, The University of Melbourne, Australia 4"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"291","published-online":{"date-parts":[[2021,1,19]]},"reference":[{"key":"2023072715422821000_bib1","doi-asserted-by":"publisher","first-page":"403","DOI":"10.1038\/87704","article-title":"Constitutive pre-TCR signaling promotes differentiation through Ca2+ mobilization and activation of NF-kappaB and NFAT","volume":"2","author":"Aifantis","year":"2001","journal-title":"Nat. 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