{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T19:36:55Z","timestamp":1768505815701,"version":"3.49.0"},"posted":{"date-parts":[[2023,1,31]]},"group-title":"Cell Biology","reference-count":73,"publisher":"openRxiv","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2023,1,31]]},"abstract":"<jats:label>1<\/jats:label>\n                <jats:title>ABSTRACT<\/jats:title>\n                <jats:p>In the central nervous system (CNS), the crosstalk between neural cells is mediated by extracellular mechanisms, including brain-derived extracellular vesicles (bdEVs).<\/jats:p>\n                <jats:p>\n                  To study endogenous communication across the brain and periphery, we explored Cre-mediated DNA recombination to permanently record the functional uptake of bdEVs cargo overtime. To elucidate functional cargo transfer within the brain at physiological levels, we promoted the continuous secretion of physiological levels of neural bdEVs containing Cre mRNA from a localized region in the brain by\n                  <jats:italic>in situ<\/jats:italic>\n                  lentiviral transduction of the striatum of Flox-tdTomato Ai9 mice reporter of Cre activity. Our approach efficiently detected in vivo transfer of functional events mediated by physiological levels of endogenous bdEVs throughout the brain. Remarkably, a spatial gradient of persistent tdTomato expression was observed along the whole brain exhibiting an increment of more than 10-fold over 4 months. Moreover, bdEVs containing Cre mRNA were detected in the bloodstream and extracted from brain tissue to further confirm their functional delivery of Cre mRNA in a novel and highly sensitive Nanoluc reporter system.\n                <\/jats:p>\n                <jats:p>Overall, we report a sensitive method to track bdEVs transfer at physiological levels which will shed light on the role of bdEVs in neural communication within the brain and beyond.<\/jats:p>","DOI":"10.1101\/2023.01.29.525937","type":"posted-content","created":{"date-parts":[[2023,1,31]],"date-time":"2023-01-31T20:20:27Z","timestamp":1675196427000},"source":"Crossref","is-referenced-by-count":1,"title":["Extracellular communication between brain cells through functional transfer of Cre mRNA"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5125-9667","authenticated-orcid":false,"given":"David","family":"Rufino-Ramos","sequence":"first","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3404-5138","authenticated-orcid":false,"given":"Kevin","family":"Leandro","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2943-3515","authenticated-orcid":false,"given":"Pedro R.L.","family":"Perdig\u00e3o","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4340-6858","authenticated-orcid":false,"given":"Killian","family":"O\u2019Brien","sequence":"additional","affiliation":[]},{"given":"Maria Manuel","family":"Pinto","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4076-7516","authenticated-orcid":false,"given":"Magda M.","family":"Santana","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3762-3618","authenticated-orcid":false,"given":"Thomas S","family":"van Solinge","sequence":"additional","affiliation":[]},{"given":"Shadi","family":"Mahjoum","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6036-0399","authenticated-orcid":false,"given":"Xandra O","family":"Breakefield","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0713-2407","authenticated-orcid":false,"given":"Koen","family":"Breyne","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5831-3307","authenticated-orcid":false,"given":"Lu\u00eds","family":"Pereira de Almeida","sequence":"additional","affiliation":[]}],"member":"54368","reference":[{"issue":"12","key":"2023020202101173000_2023.01.29.525937v1.1","doi-asserted-by":"crossref","first-page":"3105","DOI":"10.1016\/j.celrep.2019.08.036","article-title":"Glioblastoma-Associated Microglia Reprogramming Is Mediated by Functional Transfer of Extracellular MiR-21","volume":"28","year":"2019","journal-title":"Cell Reports"},{"issue":"7","key":"2023020202101173000_2023.01.29.525937v1.2","doi-asserted-by":"crossref","first-page":"393","DOI":"10.1016\/j.trecan.2019.05.006","article-title":"Glioma EVs Contribute to Immune Privilege in the Brain","volume":"5","year":"2019","journal-title":"Trends in Cancer"},{"issue":"12","key":"2023020202101173000_2023.01.29.525937v1.3","doi-asserted-by":"crossref","first-page":"e1009951","DOI":"10.1371\/journal.pgen.1009951","article-title":"MicroRNAs Are Minor Constituents of Extracellular Vesicles That Are Rarely Delivered to Target Cells","volume":"17","year":"2021","journal-title":"PLOS Genetics"},{"issue":"1","key":"2023020202101173000_2023.01.29.525937v1.4","doi-asserted-by":"crossref","first-page":"206","DOI":"10.1002\/mds.28305","article-title":"TAF1 Transcripts and Neurofilament Light Chain as Biomarkers for X-Linked Dystonia-Parkinsonism","volume":"36","year":"2021","journal-title":"Movement disorders\u202f: official journal of the Movement Disorder Society"},{"key":"2023020202101173000_2023.01.29.525937v1.5","first-page":"1","article-title":"\u201cBiomarker Potential of Brain-Secreted Extracellular Vesicles in Blood in Alzheimer\u2019s Disease","volume":"12","year":"2020","journal-title":"Alzheimer\u2019s and Dementia: Diagnosis, Assessment and Disease Monitoring"},{"key":"2023020202101173000_2023.01.29.525937v1.6","doi-asserted-by":"crossref","first-page":"180","DOI":"10.1038\/ncomms1180","article-title":"Tumour Microvesicles Contain Retrotransposon Elements and Amplified Oncogene Sequences","volume":"2","year":"2011","journal-title":"Nature communications"},{"issue":"8","key":"2023020202101173000_2023.01.29.525937v1.7","doi-asserted-by":"crossref","first-page":"974","DOI":"10.1016\/j.devcel.2022.03.012","article-title":"VAP-A and Its Binding Partner CERT Drive Biogenesis of RNA-Containing Extracellular Vesicles at ER Membrane Contact Sites","volume":"57","year":"2022","journal-title":"Developmental Cell"},{"issue":"1","key":"2023020202101173000_2023.01.29.525937v1.8","doi-asserted-by":"crossref","first-page":"15297","DOI":"10.1038\/s41598-017-15717-7","article-title":"Ultrafiltration Combined with Size Exclusion Chromatography Efficiently Isolates Extracellular Vesicles from Cell Culture Media for Compositional and Functional Studies","volume":"7","year":"2017","journal-title":"Scientific reports"},{"issue":"1","key":"2023020202101173000_2023.01.29.525937v1.9","doi-asserted-by":"crossref","first-page":"442","DOI":"10.1038\/s41467-019-08390-z","article-title":"The Functional Synergism of MicroRNA Clustering Provides Therapeutically Relevant Epigenetic Interference in Glioblastoma","volume":"10","year":"2019","journal-title":"Nature communications"},{"key":"2023020202101173000_2023.01.29.525937v1.10","doi-asserted-by":"crossref","first-page":"e10","DOI":"10.1038\/mtna.2011.2","article-title":"MiR-1289 and \u2018Zipcode\u2019-like Sequence Enrich MRNAs in Microvesicles","volume":"1","year":"2012","journal-title":"Molecular Therapy - Nucleic Acids"},{"key":"2023020202101173000_2023.01.29.525937v1.11","first-page":"1","article-title":"Exogenous Loading of Extracellular Vesicles, Virus-like Particles, and Lentiviral Vectors with Supercharged Proteins","volume":"5","year":"2022","journal-title":"Communications Biology"},{"key":"2023020202101173000_2023.01.29.525937v1.12"},{"issue":"4","key":"2023020202101173000_2023.01.29.525937v1.13","doi-asserted-by":"crossref","first-page":"1038","DOI":"10.1016\/j.ymthe.2017.01.021","article-title":"Unravelling Endogenous MicroRNA System Dysfunction as a New Pathophysiological Mechanism in Machado-Joseph Disease","volume":"25","year":"2017","journal-title":"Molecular Therapy"},{"key":"2023020202101173000_2023.01.29.525937v1.14","doi-asserted-by":"crossref","first-page":"24722","DOI":"10.3402\/jev.v3.24722","article-title":"Exosomes Secreted by Cortical Neurons upon Glutamatergic Synapse Activation Specifically Interact with Neurons","volume":"3","year":"2014","journal-title":"Journal of extracellular vesicles"},{"key":"2023020202101173000_2023.01.29.525937v1.15","first-page":"826697","article-title":"Astrocytic MicroRNA in Ageing, Inflammation, and Neurodegenerative Disease","volume":"12","year":"2021","journal-title":"Frontiers in physiology"},{"key":"2023020202101173000_2023.01.29.525937v1.16","doi-asserted-by":"publisher","DOI":"10.1038\/s41434-020-00193-y"},{"key":"2023020202101173000_2023.01.29.525937v1.17","doi-asserted-by":"crossref","unstructured":"Van Duyne, Gregory D. 2015. \u201cCre Recombinase.\u201d Microbiology spectrum 3(1): MDNA3-0014\u20132014.","DOI":"10.1128\/microbiolspec.MDNA3-0014-2014"},{"key":"2023020202101173000_2023.01.29.525937v1.18","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.5699-09.2010"},{"key":"2023020202101173000_2023.01.29.525937v1.19","doi-asserted-by":"publisher","DOI":"10.1016\/j.physbeh.2017.03.040"},{"key":"2023020202101173000_2023.01.29.525937v1.20","doi-asserted-by":"publisher","DOI":"10.1111\/j.1600-0854.2007.00677.x"},{"key":"2023020202101173000_2023.01.29.525937v1.21","first-page":"7","article-title":"Neurotransmitter-Triggered Transfer of Exosomes Mediates Oligodendrocyte-Neuron Communication","volume":"11","year":"2013","journal-title":"PLoS Biology"},{"key":"2023020202101173000_2023.01.29.525937v1.22","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-021-04234-3"},{"key":"2023020202101173000_2023.01.29.525937v1.23","doi-asserted-by":"crossref","first-page":"113961","DOI":"10.1016\/j.addr.2021.113961","article-title":"Dosing Extracellular Vesicles","volume":"178","year":"2021","journal-title":"Advanced Drug Delivery Reviews"},{"issue":"11","key":"2023020202101173000_2023.01.29.525937v1.24","doi-asserted-by":"crossref","first-page":"1848","DOI":"10.1021\/cb3002478","article-title":"Engineered Luciferase Reporter from a Deep Sea Shrimp Utilizing a Novel Imidazopyrazinone Substrate","volume":"7","year":"2012","journal-title":"ACS Chemical Biology"},{"key":"2023020202101173000_2023.01.29.525937v1.25","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.0147-18.2019"},{"key":"2023020202101173000_2023.01.29.525937v1.26","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2009.01.019"},{"key":"2023020202101173000_2023.01.29.525937v1.27","first-page":"1","article-title":"Influence of Species and Processing Parameters on Recovery and Content of Brain Tissue-Derived Extracellular Vesicles","volume":"9","year":"2020","journal-title":"Journal of Extracellular Vesicles"},{"key":"2023020202101173000_2023.01.29.525937v1.28","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2019.02.029"},{"key":"2023020202101173000_2023.01.29.525937v1.29","first-page":"5","article-title":"Intercellular Communication in the Brain through Tunneling Nanotubes","volume":"14","year":"2022","journal-title":"Cancers"},{"key":"2023020202101173000_2023.01.29.525937v1.30","first-page":"1","article-title":"Designer Exosomes Produced by Implanted Cells Intracerebrally Deliver Therapeutic Cargo for Parkinson\u2019s Disease Treatment","volume":"9","year":"2018","journal-title":"Nature Communications"},{"key":"2023020202101173000_2023.01.29.525937v1.31","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0036248"},{"key":"2023020202101173000_2023.01.29.525937v1.32","doi-asserted-by":"publisher","DOI":"10.1098\/rstb.2013.0513"},{"key":"2023020202101173000_2023.01.29.525937v1.33","doi-asserted-by":"publisher","DOI":"10.1038\/nri.2017.125"},{"issue":"4","key":"2023020202101173000_2023.01.29.525937v1.34","doi-asserted-by":"crossref","first-page":"e12206","DOI":"10.1002\/jev2.12206","article-title":"Analysis of Extracellular Vesicle DNA at the Single-Vesicle Level by Nano-Flow Cytometry","volume":"11","year":"2022","journal-title":"Journal of extracellular vesicles"},{"key":"2023020202101173000_2023.01.29.525937v1.35","doi-asserted-by":"publisher","DOI":"10.1038\/nn.2467"},{"key":"2023020202101173000_2023.01.29.525937v1.36","doi-asserted-by":"crossref","unstructured":"Mahjoum, Shadi , David Rufino-ramos , and Marike L D Broekman . 2021. \u201cLiving Proof of Activity of Extracellular Vesicles in the Central Nervous System.\u201d","DOI":"10.3390\/ijms22147294"},{"key":"2023020202101173000_2023.01.29.525937v1.37","first-page":"1","article-title":"Exosome Reporter Mice Reveal the Involvement of Exosomes in Mediating Neuron to Astroglia Communication in the CNS","volume":"10","year":"2019","journal-title":"Nature Communications"},{"key":"2023020202101173000_2023.01.29.525937v1.38","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M112.410944"},{"key":"2023020202101173000_2023.01.29.525937v1.39","doi-asserted-by":"publisher","DOI":"10.1038\/nrm.2017.125"},{"key":"2023020202101173000_2023.01.29.525937v1.40","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gks658"},{"issue":"6","key":"2023020202101173000_2023.01.29.525937v1.41","doi-asserted-by":"crossref","first-page":"631","DOI":"10.1038\/s41592-021-01174-8","article-title":"L1CAM Is Not Associated with Extracellular Vesicles in Human Cerebrospinal Fluid or Plasma","volume":"18","year":"2021","journal-title":"Nature Methods"},{"key":"2023020202101173000_2023.01.29.525937v1.42","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0012413"},{"key":"2023020202101173000_2023.01.29.525937v1.43","doi-asserted-by":"publisher","DOI":"10.1038\/s41580-020-0251-y"},{"issue":"MAY","key":"2023020202101173000_2023.01.29.525937v1.44","first-page":"1","article-title":"Lentiviral Vectors as Tools to Understand Central Nervous System Biology in Mammalian Model Organisms","volume":"8","year":"2015","journal-title":"Frontiers in Molecular Neuroscience"},{"key":"2023020202101173000_2023.01.29.525937v1.45","doi-asserted-by":"publisher","DOI":"10.1038\/onc.2015.358"},{"key":"2023020202101173000_2023.01.29.525937v1.46","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0914843107"},{"key":"2023020202101173000_2023.01.29.525937v1.47","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0603838103"},{"key":"2023020202101173000_2023.01.29.525937v1.48","first-page":"6","volume":"12","year":"2014","journal-title":"Extracellular Vesicle-Mediated Transfer of Genetic Information between the Hematopoietic System and the Brain in Response to Inflammation"},{"issue":"6","key":"2023020202101173000_2023.01.29.525937v1.49","first-page":"1","article-title":"Extracellular Vesicle-Mediated Transfer of Functional RNA in the Tumor Microenvironment","volume":"4","year":"2015","journal-title":"OncoImmunology"},{"key":"2023020202101173000_2023.01.29.525937v1.50","doi-asserted-by":"crossref","first-page":"121366","DOI":"10.1016\/j.biomaterials.2022.121366","article-title":"Biomaterials Using Genetically Modified Extracellular Vesicles as a Non-Invasive Strategy to Evaluate Brain-Specific Cargo","volume":"281","year":"2022","journal-title":"Biomaterials"},{"key":"2023020202101173000_2023.01.29.525937v1.51","unstructured":"Ruivo, Carolina F et al. 2022. \u201cExtracellular Vesicles from Pancreatic Cancer Stem Cells Lead an Intratumor Communication Network (EVNet) to Fuel Tumour Progression.\u201d Gut: gutjnl-2021\u2013324994."},{"issue":"5660","key":"2023020202101173000_2023.01.29.525937v1.52","doi-asserted-by":"crossref","first-page":"1007","DOI":"10.1126\/science.1093133","article-title":"Nanotubular Highways for Intercellular Organelle Transport","volume":"303","year":"2004","journal-title":"Science (New York, N.Y.)"},{"key":"2023020202101173000_2023.01.29.525937v1.53","doi-asserted-by":"crossref","first-page":"96","DOI":"10.1016\/j.pneurobio.2019.01.005","article-title":"New Windows into the Brain: Central Nervous System-Derived Extracellular Vesicles in Blood","volume":"175","year":"2019","journal-title":"Progress in Neurobiology"},{"key":"2023020202101173000_2023.01.29.525937v1.54","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.19276"},{"key":"2023020202101173000_2023.01.29.525937v1.55","doi-asserted-by":"publisher","DOI":"10.15252\/embj.201798099"},{"key":"2023020202101173000_2023.01.29.525937v1.56","doi-asserted-by":"crossref","unstructured":"Sterzenbach, Ulrich et al. 2017. \u201cEngineered Exosomes as Vehicles for Biologically Active Proteins.\u201d Molecular Therapy.","DOI":"10.1016\/j.ymthe.2017.03.030"},{"issue":"1","key":"2023020202101173000_2023.01.29.525937v1.57","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1186\/1479-5876-10-5","article-title":"Identification and Proteomic Profiling of Exosomes in Human Cerebrospinal Fluid","volume":"10","year":"2012","journal-title":"Journal of Translational Medicine"},{"key":"2023020202101173000_2023.01.29.525937v1.58","first-page":"7","article-title":"Characterization of Brain-Derived Extracellular Vesicle Lipids in Alzheimer\u2019s Disease","volume":"10","year":"2021","journal-title":"Journal of Extracellular Vesicles"},{"key":"2023020202101173000_2023.01.29.525937v1.59","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.62047"},{"key":"2023020202101173000_2023.01.29.525937v1.60","first-page":"1","article-title":"Minimal Information for Studies of Extracellular Vesicles 2018 (MISEV2018): A Position Statement of the International Society for Extracellular Vesicles and Update of the MISEV2014 Guidelines","volume":"7","year":"2018","journal-title":"Journal of Extracellular Vesicles"},{"key":"2023020202101173000_2023.01.29.525937v1.61","first-page":"11","article-title":"Formation of a Protein Corona on the Surface of Extracellular Vesicles in Blood Plasma","volume":"10","year":"2021","journal-title":"Journal of Extracellular Vesicles"},{"key":"2023020202101173000_2023.01.29.525937v1.62","doi-asserted-by":"publisher","DOI":"10.1038\/ncb1596"},{"key":"2023020202101173000_2023.01.29.525937v1.63","doi-asserted-by":"publisher","DOI":"10.3390\/cells9071737"},{"key":"2023020202101173000_2023.01.29.525937v1.64","first-page":"1","article-title":"A Rigorous Method to Enrich for Exosomes from Brain Tissue","volume":"6","year":"2017","journal-title":"Journal of Extracellular Vesicles"},{"issue":"1","key":"2023020202101173000_2023.01.29.525937v1.65","doi-asserted-by":"crossref","first-page":"2980","DOI":"10.1038\/ncomms3980","article-title":"Sumoylated HnRNPA2B1 Controls the Sorting of MiRNAs into Exosomes through Binding to Specific Motifs","volume":"4","year":"2013","journal-title":"Nature Communications"},{"key":"2023020202101173000_2023.01.29.525937v1.66","doi-asserted-by":"publisher","DOI":"10.1093\/neuonc\/nov244"},{"issue":"1","key":"2023020202101173000_2023.01.29.525937v1.67","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1186\/s13024-016-0143-y","article-title":"The Release and Trans-Synaptic Transmission of Tau via Exosomes","volume":"12","year":"2017","journal-title":"Molecular neurodegeneration"},{"issue":"1","key":"2023020202101173000_2023.01.29.525937v1.68","doi-asserted-by":"crossref","first-page":"1145","DOI":"10.1038\/s41467-017-01196-x","article-title":"Coding and Noncoding Landscape of Extracellular RNA Released by Human Glioma Stem Cells","volume":"8","year":"2017","journal-title":"Nature Communications"},{"issue":"1","key":"2023020202101173000_2023.01.29.525937v1.69","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1016\/j.neulet.2012.09.059","article-title":"Novel Disease-Specific Promoters for Use in Gene Therapy for Parkinson\u2019s Disease","volume":"530","year":"2012","journal-title":"Neuroscience Letters"},{"key":"2023020202101173000_2023.01.29.525937v1.70","first-page":"1","article-title":"Human Neural Cell Type-specific Extracellular Vesicle Proteome Defines Disease-related Molecules Associated with Activated Astrocytes in Alzheimer\u2019s Disease Brain","volume":"11","year":"2022","journal-title":"Journal of Extracellular Vesicles"},{"key":"2023020202101173000_2023.01.29.525937v1.71","doi-asserted-by":"publisher","DOI":"10.1172\/JCI81134"},{"key":"2023020202101173000_2023.01.29.525937v1.72","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2015.04.042"},{"issue":"1","key":"2023020202101173000_2023.01.29.525937v1.73","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1038\/nprot.2015.138","article-title":"Studying Extracellular Vesicle Transfer by a Cre-LoxP Method","volume":"11","year":"2016","journal-title":"Nature Protocols"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1101\/2023.01.29.525937","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T04:12:27Z","timestamp":1768450347000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.1101\/2023.01.29.525937"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,1,31]]},"references-count":73,"URL":"https:\/\/doi.org\/10.1101\/2023.01.29.525937","relation":{},"subject":[],"published":{"date-parts":[[2023,1,31]]},"subtype":"preprint"}}