---
title: "Endothelial Ca2+ Waves Spread Widely While Phosphatidylserine Exposure Stays Local after Vascular"
id: "biorxiv-1-spatial-uncoupling-of-endothelial-calcium-signaling-from-phospha-tidylserine"
canonical_url: "https://medichelpline.com/clinical-feed/biorxiv-1-spatial-uncoupling-of-endothelial-calcium-signaling-from-phospha-tidylserine"
content_type: "clinical_feed_article"
specialty: "Hematology"
source_name: "bioRxiv (Biomedical Preprints)"
source_url: "https://www.biorxiv.org/content/10.64898/2026.10.22.760917v1?rss=1"
published_at: "2026-10-23T12:00:00.000Z"
evidence_level: "Verified Feed"
license: "CC-BY-NC-4.0 / Informational Use"
---
# Endothelial Ca2+ Waves Spread Widely While Phosphatidylserine Exposure Stays Local after Vascular
## Provenance & Clinical Metadata
- **Canonical URL:** https://medichelpline.com/clinical-feed/biorxiv-1-spatial-uncoupling-of-endothelial-calcium-signaling-from-phospha-tidylserine
- **Specialty:** [Hematology](https://medichelpline.com/clinical-feed/hematology.md)
- **Primary Source:** bioRxiv (Biomedical Preprints)
- **Source URL:** [Original Journal Publication](https://www.biorxiv.org/content/10.64898/2026.10.22.760917v1?rss=1)
- **Published At:** 2026-10-23T12:00:00.000Z
- **Evidence Rating:** Verified Feed
## Executive GIST (TL;DR)
- Investigators combined single-cell laser injury in endothelial monolayers, imaging of intact mesenteric vessels in vitro, and intravital Ca2+ imaging in vivo to probe injury signaling and membrane scrambling. - A coherent **calcium** (Ca2+) wavefront spread rapidly from the wound then slowed and stopped at a reproducible boundary 197 ± 7 μm from the injury, engaging 50 ± 8 cells on average. - The immediate neighbor ring of cells produced a distinct response that was blocked by carbenoxolone, whereas propagation beyond that ring required extracellular ATP, purinergic receptor activation, and IP3-dependent Ca2+ release. - Wavefront kinematics matched expectations for ATP diffusion as the extracellular messenger driving farther propagation. - Intravital experiments showed injury triggered vasoconstriction and bidirectional Ca2+ waves that propagated 205 ± 28 μm before terminating. - Despite broad endothelial Ca2+ elevations, phosphatidylserine (PS) externalization remained confined to the wounded cell in monolayers and in intact vessels in vitro. - By contrast, ionomycin-induced global Ca2+ overload exposed PS across nearly all endothelial cells, showing that generalized Ca2+ elevation can drive scrambling when unregulated. - Authors concluded that Ca2+-dependent scramblase activation is subject to additional spatial regulation, permitting injury signal spread without creating a wide procoagulant endothelial surface. - The source did not report detailed limitations of the experimental models or specific molecular identity of the scramblase(s) responsible, nor did it provide clinical translation steps or dosing regimens for modulators beyond experimental inhibitors noted.
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Nelson, [ View ORCID Profile](http://orcid.org/0000-0003-4817-2955)Kalev Freeman doi: https://doi.org/10.64898/2026.10.22.760917 This article is a preprint and has not been certified by peer review [[what does this mean?](https://www.biorxiv.org/about/FAQ#unrefereed)]. Olivia Fauver 1 University of Vermont; * [Find this author on Google Scholar](https://www.biorxiv.org/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Olivia%2BFauver%2B "Open in new tab") * [Find this author on PubMed](https://www.biorxiv.org/lookup/external-ref?access_num=Fauver%20O&link_type=AUTHORSEARCH "Open in new tab") * [Search for this author on this site](https://www.biorxiv.org/search/author1%3AOlivia%2BFauver%2B) Grant Hennig 1 University of Vermont; * [Find this author on Google Scholar](https://www.biorxiv.org/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Grant%2BHennig%2B "Open in new tab") * [Find this author on PubMed](https://www.biorxiv.org/lookup/external-ref?access_num=Hennig%20G&link_type=AUTHORSEARCH "Open in new tab") * [Search for this author on this site](https://www.biorxiv.org/search/author1%3AGrant%2BHennig%2B) * [ORCID record for Grant Hennig](http://orcid.org/0000-0001-7436-9458 "Open in new tab") Adrian Sackheim 1 University of Vermont; * [Find this author on Google Scholar](https://www.biorxiv.org/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Adrian%2BSackheim%2B "Open in new tab") * [Find this author on PubMed](https://www.biorxiv.org/lookup/external-ref?access_num=Sackheim%20A&link_type=AUTHORSEARCH "Open in new tab") * [Search for this author on this site](https://www.biorxiv.org/search/author1%3AAdrian%2BSackheim%2B) Bryan Walters 1 University of Vermont; * [Find this author on Google Scholar](https://www.biorxiv.org/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Bryan%2BWalters%2B "Open in new tab") * [Find this author on PubMed](https://www.biorxiv.org/lookup/external-ref?access_num=Walters%20B&link_type=AUTHORSEARCH "Open in new tab") * [Search for this author on this site](https://www.biorxiv.org/search/author1%3ABryan%2BWalters%2B) Rachel Barba 1 University of Vermont; * [Find this author on Google Scholar](https://www.biorxiv.org/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Rachel%2BBarba%2B "Open in new tab") * [Find this author on PubMed](https://www.biorxiv.org/lookup/external-ref?access_num=Barba%20R&link_type=AUTHORSEARCH "Open in new tab") * [Search for this author on this site](https://www.biorxiv.org/search/author1%3ARachel%2BBarba%2B) Rushi Sonawane 1 University of Vermont; * [Find this author on Google Scholar](https://www.biorxiv.org/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Rushi%2BSonawane%2B "Open in new tab") * [Find this author on PubMed](https://www.biorxiv.org/lookup/external-ref?access_num=Sonawane%20R&link_type=AUTHORSEARCH "Open in new tab") * [Search for this author on this site](https://www.biorxiv.org/search/author1%3ARushi%2BSonawane%2B) Christopher Bidlack 1 University of Vermont; * [Find this author on Google Scholar](https://www.biorxiv.org/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Christopher%2BBidlack%2B "Open in new tab") * [Find this author on PubMed](https://www.biorxiv.org/lookup/external-ref?access_num=Bidlack%20C&link_type=AUTHORSEARCH "Open in new tab") * [Search for this author on this site](https://www.biorxiv.org/search/author1%3AChristopher%2BBidlack%2B) Devdoot Majumdar 1 University of Vermont; * [Find this author on Google Scholar](https://www.biorxiv.org/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Devdoot%2BMajumdar%2B "Open in new tab") * [Find this author on PubMed](https://www.biorxiv.org/lookup/external-ref?access_num=Majumdar%20D&link_type=AUTHORSEARCH "Open in new tab") * [Search for this author on this site](https://www.biorxiv.org/search/author1%3ADevdoot%2BMajumdar%2B) * [ORCID record for Devdoot Majumdar](http://orcid.org/0000-0002-2541-3851 "Open in new tab") Mohamed Trebak 2 University of Pittsburgh * [Find this author on Google Scholar](https://www.biorxiv.org/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Mohamed%2BTrebak%2B "Open in new tab") * [Find this author on PubMed](https://www.biorxiv.org/lookup/external-ref?access_num=Trebak%20M&link_type=AUTHORSEARCH "Open in new tab") * [Search for this author on this site](https://www.biorxiv.org/search/author1%3AMohamed%2BTrebak%2B) * [ORCID record for Mohamed Trebak](http://orcid.org/0000-0001-6759-864X "Open in new tab") Mark T. Nelson 1 University of Vermont; * [Find this author on Google Scholar](https://www.biorxiv.org/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Mark%2BT.%2BNelson%2B "Open in new tab") * [Find this author on PubMed](https://www.biorxiv.org/lookup/external-ref?access_num=Nelson%20MT&link_type=AUTHORSEARCH "Open in new tab") * [Search for this author on this site](https://www.biorxiv.org/search/author1%3AMark%2BT.%2BNelson%2B) Kalev Freeman 1 University of Vermont; * [Find this author on Google Scholar](https://www.biorxiv.org/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Kalev%2BFreeman%2B "Open in new tab") * [Find this author on PubMed](https://www.biorxiv.org/lookup/external-ref?access_num=Freeman%20K&link_type=AUTHORSEARCH "Open in new tab") * [Search for this author on this site](https://www.biorxiv.org/search/author1%3AKalev%2BFreeman%2B) * [ORCID record for Kalev Freeman](http://orcid.org/0000-0003-4817-2955 "Open in new tab") * For correspondence: kalev.freeman@uvm.edu * [Abstract](https://www.biorxiv.org/content/10.64898/2026.10.22.760917v1)[](https://www.biorxiv.org/panels_ajax_tab/biorxiv_tab_art/node:5895871/1) * [Info/History](https://www.biorxiv.org/content/10.64898/2026.10.22.760917v1.article-info)[](https://www.biorxiv.org/panels_ajax_tab/biorxiv_tab_info/node:5895871/1) * [Metrics](https://www.biorxiv.org/content/10.64898/2026.10.22.760917v1.article-metrics)[](https://www.biorxiv.org/panels_ajax_tab/article_tab_metrics/node:5895871/1) * [Supplementary material](https://www.biorxiv.org/content/10.64898/2026.10.22.760917v1.supplementary-material)[](https://www.biorxiv.org/panels_ajax_tab/biorxiv_tab_data/node:5895871/1) * [ Preview PDF](https://www.biorxiv.org/content/10.64898/2026.10.22.760917v1.full.pdf+html)[](https://www.biorxiv.org/panels_ajax_tab/biorxiv_tab_pdf/node:5895871/1) ![Loading](https://www.biorxiv.org/sites/all/modules/contrib/panels_ajax_tab/images/loading.gif) ## Abstract Ca2+ is both the signal that spreads injury information through the endothelium and the trigger for phospholipid scrambling that makes a membrane procoagulant. How a blood vessel can disseminate the first without broadcasting the second is unknown. Here, we combine single-cell laser injury in endothelial monolayers with imaging of intact mesenteric vessels in vitro and intravital Ca2+ imaging in vivo. Injury generated a coherent Ca2+ wavefront that spread rapidly before progressively slowing and terminating at a reproducible boundary 197 ± 7 μm from the wound, engaging 50 ± 8 cells. The first ring of neighboring cells exhibited a distinct response that was prevented by carbenoxolone, whereas propagation beyond this ring required extracellular ATP, purinergic receptor activation, and IP3-dependent Ca2+ release. Wavefront kinematics were consistent with ATP diffusion. In vivo, injury elicited vasoconstriction and bidirectional Ca2+ waves propagating 205 ± 28 μm before terminating. Remarkably, despite widespread Ca2+ elevations, phosphatidylserine (PS) externalization remained confined to the wounded cell in monolayers and intact vessels in vitro. In contrast, ionomycin-induced Ca2+ overload exposed phosphatidylserine in nearly all endothelial cells. These findings reveal an unexpected spatial uncoupling between endothelial Ca2+ signaling and phospholipid scrambling, demonstrating that Ca2+ dependent scramblase activation is subject to additional spatial regulation. The endothelium can therefore propagate injury signals across a broad field of cells while maintaining a highly localized procoagulant membrane response. ### Competing Interest Statement The authors have declared no competing interest. ## Funder Information Declared National Institute of General Medical Sciences, https://ror.org/04q48ey07, R35GM144099 Copyright The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a [CC-BY 4.0 International license](http://creativecommons.org/licenses/by/4.0/). bioRxiv and medRxiv thank the following for their generous financial support: > The Chan Zuckerberg Initiative, Cold Spring Harbor Laboratory, the Sergey Brin Family Foundation, California Institute of Technology, Centre National de la Recherche Scientifique, Fred Hutchinson Cancer Center, Imperial College London, Massachusetts Institute of Technology, Stanford University, The University of Edinburgh, University of Washington, and Vrije Universiteit Amsterdam. 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[ Download PDF](https://www.biorxiv.org/content/10.64898/2026.10.22.760917v1.full.pdf) Print/Save Options [Download PDF](https://www.biorxiv.org/content/biorxiv/early/2026/10/23/2026.10.22.760917.full.pdf)Full Text & In-line FiguresXML [More Info](https://www.biorxiv.org/about/FAQ#PrintOptions "More Information on Print/Save Options") [Supplementary Material ](https://www.biorxiv.org/content/10.64898/2026.10.22.760917v1.supplementary-material) [ Email](https://www.biorxiv.org/ "Email this Article") [ Share](https://www.biorxiv.org/) Spatial uncoupling of endothelial calcium signaling from phospha-tidylserine externalization in vascular injury Olivia Fauver, Grant Hennig, Adrian Sackheim, Bryan Walters, Rachel Barba, Rushi Sonawane, Christopher Bidlack, Devdoot Majumdar, Mohamed Trebak, Mark T. Nelson, Kalev Freeman bioRxiv 2026.10.22.760917; doi: https://doi.org/10.64898/2026.10.22.760917 This article is a preprint and has not been certified by peer review [[what does this mean?](https://www.biorxiv.org/about/FAQ#unrefereed)]. Share This Article: Copy [![Twitter logo](https://www.biorxiv.org/sites/all/modules/highwire/highwire/images/twitter.png)](https://www.biorxiv.org/highwire_lo
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