{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T16:04:46Z","timestamp":1788365086600,"version":"build-2803163510"},"posted":{"date-parts":[[2025,4,7]]},"group-title":"Molecular Biology","reference-count":78,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,4,7]],"date-time":"2025-04-07T00:00:00Z","timestamp":1743984000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,4,7]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>The blood-brain barrier (BBB) glycocalyx is the dense layer of glycans and glycoconjugates that coats the luminal surface of the central nervous system (CNS) vasculature. Despite being the first point of contact between the blood and brain, not much is known about the BBB glycocalyx. Here, we performed a multi-omic investigation of the BBB glycocalyx which revealed a unique glycan landscape characterized by enrichment of sialic acid, chondroitin sulfate, and hyaluronan. We found that the BBB glycocalyx was thicker than glycocalyces in the peripheral vasculature and that hyaluronan was the major contributor to its ultrastructure. Using endothelial RNA sequencing, we found potential genetic determinants for these differences, including BBB enrichment of genes involved in sialic acid addition and peripheral enrichment of Tmem2 and Hyal2, the only known cell-surface hyaluronidases. Glycocalyx degradation and increases in vascular permeability are widely associated with inflammation. However, we found that the BBB glycocalyx remains largely unchanged in neuroinflammation during the experimental autoimmune encephalomyelitis (EAE) model of multiple sclerosis and that its degradation is not sufficient to alter BBB permeability in health. Moreover, we showed that CNS endothelial sialic acid removal delays onset of EAE, indicating that BBB glycocalyx sialic acid may contribute to the progression of neuroinflammation. These findings underscore the unique and robust nature of the BBB glycocalyx and provide targets and tools for future studies into its role in health and neuroinflammation.<\/jats:p>","DOI":"10.1101\/2025.04.07.645297","type":"posted-content","created":{"date-parts":[[2025,4,7]],"date-time":"2025-04-07T12:40:58Z","timestamp":1744029658000},"source":"Crossref","is-referenced-by-count":9,"title":["Multi-omic analysis reveals the unique glycan landscape of the blood-brain barrier glycocalyx"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9783-636X","authenticated-orcid":false,"given":"Reid","family":"Larsen","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Krzysztof","family":"Kucharz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7791-7931","authenticated-orcid":false,"given":"Sidar","family":"Aydin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9656-406X","authenticated-orcid":false,"given":"Mariel Kristine B.","family":"Micael","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Biswa","family":"Choudhury","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mousumi","family":"Paulchakrabarti","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4302-7978","authenticated-orcid":false,"given":"Micael","family":"L\u00f8nstrup","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-1447-4979","authenticated-orcid":false,"given":"Ding Chiao","family":"Lin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Markus","family":"Abeln","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anja","family":"M\u00fcnster-K\u00fchnel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alejandro Gomez","family":"Toledo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martin","family":"Lauritzen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8322-1834","authenticated-orcid":false,"given":"Jeffrey D.","family":"Esko","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Richard","family":"Daneman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2025040915501882000_2025.04.07.645297v1.1","doi-asserted-by":"publisher","DOI":"10.1152\/physrev.00050.2017"},{"key":"2025040915501882000_2025.04.07.645297v1.2","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20190062"},{"key":"2025040915501882000_2025.04.07.645297v1.3","doi-asserted-by":"publisher","DOI":"10.1101\/cshperspect.a020412"},{"key":"2025040915501882000_2025.04.07.645297v1.4","doi-asserted-by":"publisher","DOI":"10.1016\/j.nbd.2009.07.030"},{"key":"2025040915501882000_2025.04.07.645297v1.5","doi-asserted-by":"publisher","DOI":"10.1186\/s12987-020-00230-3"},{"key":"2025040915501882000_2025.04.07.645297v1.6","doi-asserted-by":"crossref","unstructured":"Haeren, R. et al. Assessment and Imaging of the Cerebrovascular Glycocalyx. Current neurovascular research 13, (2016).","DOI":"10.2174\/1567202613666160504104434"},{"key":"2025040915501882000_2025.04.07.645297v1.7","doi-asserted-by":"publisher","DOI":"10.1189\/jlb.3MR0115-011R"},{"key":"2025040915501882000_2025.04.07.645297v1.8","doi-asserted-by":"publisher","DOI":"10.1002\/ams2.896"},{"key":"2025040915501882000_2025.04.07.645297v1.9","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-biochem-032620-104745"},{"key":"2025040915501882000_2025.04.07.645297v1.10","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.bioeng.9.060906.151959"},{"key":"2025040915501882000_2025.04.07.645297v1.11","doi-asserted-by":"publisher","DOI":"10.1007\/s00424-007-0212-8"},{"key":"2025040915501882000_2025.04.07.645297v1.12","doi-asserted-by":"publisher","DOI":"10.1016\/j.mvr.2010.06.005"},{"key":"2025040915501882000_2025.04.07.645297v1.13","unstructured":"Essentials of Glycobiology. (Cold Spring Harbor Laboratory Press, Cold Spring Harbor (NY), 2022)."},{"key":"2025040915501882000_2025.04.07.645297v1.14","doi-asserted-by":"publisher","DOI":"10.1111\/j.1749-6632.2012.06517.x"},{"key":"2025040915501882000_2025.04.07.645297v1.15","doi-asserted-by":"publisher","DOI":"10.1007\/s004240000307"},{"key":"2025040915501882000_2025.04.07.645297v1.16","doi-asserted-by":"publisher","DOI":"10.1113\/JP274167"},{"key":"2025040915501882000_2025.04.07.645297v1.17","doi-asserted-by":"publisher","DOI":"10.1152\/ajpheart.2000.279.6.H2815"},{"key":"2025040915501882000_2025.04.07.645297v1.18","doi-asserted-by":"publisher","DOI":"10.1016\/j.ajpath.2019.11.015"},{"key":"2025040915501882000_2025.04.07.645297v1.19","doi-asserted-by":"publisher","DOI":"10.1007\/s10439-011-0427-x"},{"key":"2025040915501882000_2025.04.07.645297v1.20","doi-asserted-by":"publisher","DOI":"10.1038\/nm.2843"},{"key":"2025040915501882000_2025.04.07.645297v1.21","doi-asserted-by":"publisher","DOI":"10.1016\/j.yexcr.2011.01.004"},{"key":"2025040915501882000_2025.04.07.645297v1.22","doi-asserted-by":"publisher","DOI":"10.1038\/ni1233"},{"key":"2025040915501882000_2025.04.07.645297v1.23","doi-asserted-by":"publisher","DOI":"10.1177\/0271678X21992638"},{"key":"2025040915501882000_2025.04.07.645297v1.24","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1802155115"},{"key":"2025040915501882000_2025.04.07.645297v1.25","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-025-08589-9"},{"key":"2025040915501882000_2025.04.07.645297v1.26","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-018-35976-2"},{"key":"2025040915501882000_2025.04.07.645297v1.27","doi-asserted-by":"publisher","DOI":"10.1177\/0271678X17726062"},{"key":"2025040915501882000_2025.04.07.645297v1.28","doi-asserted-by":"publisher","DOI":"10.1097\/TA.0000000000000333"},{"key":"2025040915501882000_2025.04.07.645297v1.29","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2018.01254"},{"key":"2025040915501882000_2025.04.07.645297v1.30","doi-asserted-by":"publisher","DOI":"10.1161\/STROKEAHA.119.025953"},{"key":"2025040915501882000_2025.04.07.645297v1.31","doi-asserted-by":"crossref","unstructured":"Rj, M. , H, V., Rj, van O. & J, S. Sublingual microvascular glycocalyx dimensions in lacunar stroke patients. Cerebrovascular diseases (Basel, Switzerland) 35, (2013).","DOI":"10.1159\/000348854"},{"key":"2025040915501882000_2025.04.07.645297v1.32","doi-asserted-by":"crossref","unstructured":"Zhu, J. et al. Glycocalyx is critical for blood-brain barrier integrity by suppressing caveolin1-dependent endothelial transcytosis following ischemic stroke. Brain Pathology n\/a, e13006 (2021).","DOI":"10.1111\/bpa.13006"},{"key":"2025040915501882000_2025.04.07.645297v1.33","doi-asserted-by":"publisher","DOI":"10.1097\/00004647-200011000-00007"},{"key":"2025040915501882000_2025.04.07.645297v1.34","doi-asserted-by":"publisher","DOI":"10.1186\/s13054-017-1841-8"},{"key":"2025040915501882000_2025.04.07.645297v1.35","doi-asserted-by":"publisher","DOI":"10.1007\/BF01011888"},{"key":"2025040915501882000_2025.04.07.645297v1.36","doi-asserted-by":"publisher","DOI":"10.1038\/s41580-020-00294-x"},{"key":"2025040915501882000_2025.04.07.645297v1.37","doi-asserted-by":"publisher","DOI":"10.1093\/glycob\/cwaa010"},{"key":"2025040915501882000_2025.04.07.645297v1.38","doi-asserted-by":"crossref","unstructured":"Patterson, E. K. , Cepinskas, G. & Fraser, D. D . Endothelial Glycocalyx Degradation in Critical Illness and Injury. Frontiers in Medicine 9, (2022).","DOI":"10.3389\/fmed.2022.898592"},{"key":"2025040915501882000_2025.04.07.645297v1.39","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-022-31062-4"},{"key":"2025040915501882000_2025.04.07.645297v1.40","doi-asserted-by":"publisher","DOI":"10.1038\/s41593-019-0497-x"},{"key":"2025040915501882000_2025.04.07.645297v1.41","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbc.2021.100481"},{"key":"2025040915501882000_2025.04.07.645297v1.42","doi-asserted-by":"publisher","DOI":"10.1016\/j.isci.2022.104419"},{"key":"2025040915501882000_2025.04.07.645297v1.43","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M414276200"},{"key":"2025040915501882000_2025.04.07.645297v1.44","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-019-12672-x"},{"key":"2025040915501882000_2025.04.07.645297v1.45","doi-asserted-by":"publisher","DOI":"10.1186\/s12987-024-00523-x"},{"key":"2025040915501882000_2025.04.07.645297v1.46","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth745"},{"key":"2025040915501882000_2025.04.07.645297v1.47","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2006.29"},{"key":"2025040915501882000_2025.04.07.645297v1.48","doi-asserted-by":"publisher","DOI":"10.1007\/s11095-022-03266-2"},{"key":"2025040915501882000_2025.04.07.645297v1.49","doi-asserted-by":"crossref","unstructured":"Zhang, Q. et al. High Level of Serum and Cerebrospinal Fluid of Heparan Sulfate and Hyaluronic Acid Might Be a Biomarker of Severity of Neuromyelitis Optica. Frontiers in Immunology 12, (2021).","DOI":"10.3389\/fimmu.2021.705536"},{"key":"2025040915501882000_2025.04.07.645297v1.50","doi-asserted-by":"publisher","DOI":"10.1016\/j.expneurol.2020.113320"},{"key":"2025040915501882000_2025.04.07.645297v1.51","doi-asserted-by":"publisher","DOI":"10.1111\/j.1476-5381.2011.01302.x"},{"key":"2025040915501882000_2025.04.07.645297v1.52","doi-asserted-by":"publisher","DOI":"10.1016\/B978-0-444-52001-2.00008-X"},{"key":"2025040915501882000_2025.04.07.645297v1.53","unstructured":"Miller, S. D. , Karpus, W. J. & Davidson, T. S . Experimental Autoimmune Encephalomyelitis in the Mouse. Curr Protoc Immunol CHAPTER, Unit-15.1 (2007)."},{"key":"2025040915501882000_2025.04.07.645297v1.54","doi-asserted-by":"publisher","DOI":"10.1016\/j.clim.2013.06.004"},{"key":"2025040915501882000_2025.04.07.645297v1.55","doi-asserted-by":"publisher","DOI":"10.1093\/glycob\/cwu101"},{"key":"2025040915501882000_2025.04.07.645297v1.56","doi-asserted-by":"publisher","DOI":"10.1002\/cbic.201700083"},{"key":"2025040915501882000_2025.04.07.645297v1.57","doi-asserted-by":"publisher","DOI":"10.1172\/JCI99945"},{"key":"2025040915501882000_2025.04.07.645297v1.58","doi-asserted-by":"publisher","DOI":"10.1111\/j.1549-8719.2012.00168.x"},{"key":"2025040915501882000_2025.04.07.645297v1.59","doi-asserted-by":"publisher","DOI":"10.1111\/micc.12643"},{"key":"2025040915501882000_2025.04.07.645297v1.60","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCRESAHA.107.160226"},{"key":"2025040915501882000_2025.04.07.645297v1.61","doi-asserted-by":"publisher","DOI":"10.1161\/ATVBAHA.111.225268"},{"key":"2025040915501882000_2025.04.07.645297v1.62","doi-asserted-by":"publisher","DOI":"10.1152\/ajpheart.2000.278.1.H285"},{"key":"2025040915501882000_2025.04.07.645297v1.63","doi-asserted-by":"publisher","DOI":"10.1152\/ajpheart.2000.279.6.H2815"},{"key":"2025040915501882000_2025.04.07.645297v1.64","doi-asserted-by":"publisher","DOI":"10.1038\/s41423-021-00757-x"},{"key":"2025040915501882000_2025.04.07.645297v1.65","doi-asserted-by":"publisher","DOI":"10.1101\/2024.05.21.595098"},{"key":"2025040915501882000_2025.04.07.645297v1.66","first-page":"1135","article-title":"Role of Syndecan-1 in Leukocyte\u2013Endothelial Interactions in the Ocular Vasculature","volume":"43","year":"2002","journal-title":"Investigative Ophthalmology & Visual Science"},{"key":"2025040915501882000_2025.04.07.645297v1.67","doi-asserted-by":"publisher","DOI":"10.1038\/jcbfm.2014.79"},{"key":"2025040915501882000_2025.04.07.645297v1.68","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-0716-2059-5_8"},{"key":"2025040915501882000_2025.04.07.645297v1.69","doi-asserted-by":"publisher","DOI":"10.1002\/JLB.3RU0618-246R"},{"key":"2025040915501882000_2025.04.07.645297v1.70","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbrc.2009.03.140"},{"key":"2025040915501882000_2025.04.07.645297v1.71","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2008-08-174508"},{"key":"2025040915501882000_2025.04.07.645297v1.72","doi-asserted-by":"publisher","DOI":"10.3389\/fnagi.2019.00289"},{"key":"2025040915501882000_2025.04.07.645297v1.73","doi-asserted-by":"publisher","DOI":"10.1021\/pr7008252"},{"key":"2025040915501882000_2025.04.07.645297v1.74","doi-asserted-by":"publisher","DOI":"10.1038\/labinvest.3700281"},{"key":"2025040915501882000_2025.04.07.645297v1.75","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.49405"},{"key":"2025040915501882000_2025.04.07.645297v1.76","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-021-24323-1"},{"key":"2025040915501882000_2025.04.07.645297v1.77","unstructured":"Li, K. The Image Stabilizer Plugin for ImageJ. https:\/\/www.cs.cmu.edu\/~kangli\/code\/Image_Stabilizer.html."},{"key":"2025040915501882000_2025.04.07.645297v1.78","doi-asserted-by":"publisher","DOI":"10.1002\/1521-4141(200207)32:7<1905::AID-IMMU1905>3.0.CO;2-L"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1101\/2025.04.07.645297","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T08:26:06Z","timestamp":1768465566000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.1101\/2025.04.07.645297"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,4,7]]},"references-count":78,"URL":"https:\/\/doi.org\/10.1101\/2025.04.07.645297","relation":{},"subject":[],"published":{"date-parts":[[2025,4,7]]},"subtype":"preprint"}}