Imaging of Neurovascular Activity during Motor Responses Evoked by Displacement-guided Focused Ultrasound (DgFUS) Neuromodulation in Mice

Imaging of Neurovascular Activity during Motor Responses Evoked by Displacement-guided Focused Ultrasound (DgFUS) Neuromodulation in Mice | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Imaging of Neurovascular Activity during Motor Responses Evoked by Displacement-guided Focused Ultrasound (DgFUS) Neuromodulation in Mice Elisa Konofagou, Seongyeon Kim, Daniella Jimenez, Jonas Bendig, and 7 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9173358/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted You are reading this latest preprint version Abstract Focused ultrasound (FUS) shows great promise for precise and non-invasive neuromodulation, with applications from basic neuroscience investigation to the treatment of neurological disorders. Yet, mechanistic monitoring of FUS is challenging and a causal link between FUS-induced displacement and both neurovascular and behavioral responses evoked by FUS in mice remains poorly understood. Here we harness displacement-guided focused ultrasound (DgFUS) to induce target-specific and non-auditory motor responses in anesthetized mice, and functional ultrasound (fUS) to image neurovascular activity during the evoked responses. We show that DgFUS elicits ipsilateral neurovascular responses, in which the primary (i.e. focal-onset) activation localizes with brain displacement, followed by mild contralateral responses. We demonstrate that switching the target hemisphere of DgFUS within the thalamus results in lateralized hindlimb responses concurrent with lateralized neurovascular activation. We show strong correlation between displacement and both cerebral blood volume (CBV) and electromyography (EMG) measurement (Disp. vs. CBV: Spearman r = 0.9091, p < 0.0001, Disp. vs. EMG: r = 0.7332, p = 0.0091, CBV vs. EMG: r = 0.9041, p = 0.0002), indicating displacement-driven mechanism by which DgFUS evokes neurovascular and motor responses. Lastly, we demonstrate anxiety-related behavioral improvement with transcranial fUS-guided FUS delivered to the bilateral hippocampus, consistent with increases in resting-state hippocampal connectivity assessed by transcranial fUS imaging. The findings presented herein will help refine our understanding of neurovascular responses to FUS and the consequential motor and cognitive outcomes. Biological sciences/Neuroscience Biological sciences/Biological techniques/Imaging/Ultrasound Displacement-guided focused ultrasound (DgFUS) Functional ultrasound (fUS) imaging Neurovascular response Cerebral blood volume (CBV) Electromyography (EMG) Full Text Additional Declarations There is NO Competing Interest. Supplementary Files SupplemantaryMovie3A.mp4 Supplemantary Movie 3A SupplementaryMovie5ERTh.mp4 Supplemantary Movie 5E SupplementaryMovie5DLTh.mp4 Supplemantary Movie 5D Supplementarydocument.docx Supplementary document SupplementaryMovie5BRHL.mp4 Supplemantary Movie 5B SupplementaryMovie5ALHL.mp4 Supplemantary Movie 5A SupplementaryMovie4ALHL.mp4 Supplemantary Movie 4A SupplementaryMovie5COfftarget.mp4 Supplemantary Movie 5C SupplementaryMovie3BToetwitch.mp4 Supplemantary Movie 3B SupplementaryMovie4BRHL.mp4 Supplemantary Movie 4B Cite Share Download PDF Status: Under Review Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-9173358","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":612341016,"identity":"16f263f9-7153-4e47-9000-feb62b2b1d11","order_by":0,"name":"Elisa Konofagou","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA3ElEQVRIie3PoQvCQBTH8SeDrTy5uhX9F56IJpn/ykQwCV6yGBQWLLNP8I8QBPONAy3iqnDdtDCwGjxBk7LNZrhvPO7D4wdgMv1jaAmRUw/AAbCAnk9BGbEHScxHz/+VCbYl5vJFoAJhKEgipU1mYWfHuQ/MGVMh8VaLINmQaq1D7KqYhuBFWTGhFITISAX6Vlch6TmXkiv9tLYQSOc3mUO/jFA9hARJvIkEckuIezpAoifoLfZUbzmie7ryQsKiyS3P736TOeFe8fuswZbDbSH5CH/7bjKZTKavPQABwUX8Y+mKsgAAAABJRU5ErkJggg==","orcid":"https://orcid.org/0000-0002-9636-7936","institution":"Columbia University","correspondingAuthor":true,"prefix":"","firstName":"Elisa","middleName":"","lastName":"Konofagou","suffix":""},{"id":612341017,"identity":"9727ae0d-97f1-4967-b92f-753f75192adf","order_by":1,"name":"Seongyeon Kim","email":"","orcid":"","institution":"Columbia University","correspondingAuthor":false,"prefix":"","firstName":"Seongyeon","middleName":"","lastName":"Kim","suffix":""},{"id":612341018,"identity":"756beec2-419b-4cce-941f-ee6f38909496","order_by":2,"name":"Daniella Jimenez","email":"","orcid":"","institution":"Columbia University","correspondingAuthor":false,"prefix":"","firstName":"Daniella","middleName":"","lastName":"Jimenez","suffix":""},{"id":612341019,"identity":"c8801b18-f762-407e-9a03-4ad6c6c16cb9","order_by":3,"name":"Jonas Bendig","email":"","orcid":"","institution":"Columbia University","correspondingAuthor":false,"prefix":"","firstName":"Jonas","middleName":"","lastName":"Bendig","suffix":""},{"id":612341020,"identity":"c11bd47f-3622-48d1-9eef-576eb3eb09f4","order_by":4,"name":"Moshe Willner","email":"","orcid":"","institution":"Columbia University","correspondingAuthor":false,"prefix":"","firstName":"Moshe","middleName":"","lastName":"Willner","suffix":""},{"id":612341021,"identity":"145baad4-27f1-4ee7-9695-554c7861d8f1","order_by":5,"name":"Gillian Ciaccio","email":"","orcid":"","institution":"Columbia University","correspondingAuthor":false,"prefix":"","firstName":"Gillian","middleName":"","lastName":"Ciaccio","suffix":""},{"id":612341022,"identity":"3ceb9b44-6541-4c04-8eb9-0bec6cb55999","order_by":6,"name":"Sergio Jimenez-Gambin","email":"","orcid":"","institution":"Columbia University","correspondingAuthor":false,"prefix":"","firstName":"Sergio","middleName":"","lastName":"Jimenez-Gambin","suffix":""},{"id":612341023,"identity":"d26e553d-37fc-403c-bed3-decdd88d4acd","order_by":7,"name":"Nancy Kwon","email":"","orcid":"","institution":"Columbia University","correspondingAuthor":false,"prefix":"","firstName":"Nancy","middleName":"","lastName":"Kwon","suffix":""},{"id":612341024,"identity":"4786eaaa-f366-4eb2-8485-c257ca284e36","order_by":8,"name":"Saachi Munot","email":"","orcid":"","institution":"Columbia University","correspondingAuthor":false,"prefix":"","firstName":"Saachi","middleName":"","lastName":"Munot","suffix":""},{"id":612341025,"identity":"50832469-2cdd-4209-b1b0-a25fb0e5ffa1","order_by":9,"name":"Erica McCune","email":"","orcid":"","institution":"Columbia University","correspondingAuthor":false,"prefix":"","firstName":"Erica","middleName":"","lastName":"McCune","suffix":""},{"id":612341026,"identity":"66524029-3182-4b71-afe9-f47cf2ac3e6b","order_by":10,"name":"Samuel Blackman","email":"","orcid":"","institution":"Columbia University","correspondingAuthor":false,"prefix":"","firstName":"Samuel","middleName":"","lastName":"Blackman","suffix":""}],"badges":[],"createdAt":"2026-03-19 23:40:19","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-9173358/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-9173358/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":108183746,"identity":"31bfdd6f-643b-4a12-b5ad-a4bd1b9c1463","added_by":"auto","created_at":"2026-04-30 09:02:38","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3830627,"visible":true,"origin":"","legend":"Article File","description":"","filename":"Paper2NatureCommSKFinal2.pdf","url":"https://assets-eu.researchsquare.com/files/rs-9173358/v1_covered_d6bcb413-179f-4f61-8eeb-18d07f70bc55.pdf"},{"id":107707614,"identity":"b88ca0c3-dde3-4fff-8c24-c0483f6dcfdc","added_by":"auto","created_at":"2026-04-24 09:20:45","extension":"mp4","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":423201,"visible":true,"origin":"","legend":"Supplemantary Movie 3A","description":"","filename":"SupplemantaryMovie3A.mp4","url":"https://assets-eu.researchsquare.com/files/rs-9173358/v1/2f8fc0f16558327180596508.mp4"},{"id":107629043,"identity":"b799f748-767a-4074-9a94-5659ca759d3e","added_by":"auto","created_at":"2026-04-23 11:19:25","extension":"mp4","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":288306,"visible":true,"origin":"","legend":"Supplemantary Movie 5E","description":"","filename":"SupplementaryMovie5ERTh.mp4","url":"https://assets-eu.researchsquare.com/files/rs-9173358/v1/f914dec4a823b21b4d71fdf8.mp4"},{"id":107707476,"identity":"e8c3bb60-c6be-4d55-b5dd-6c7222486305","added_by":"auto","created_at":"2026-04-24 09:20:24","extension":"mp4","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":305960,"visible":true,"origin":"","legend":"Supplemantary Movie 5D","description":"","filename":"SupplementaryMovie5DLTh.mp4","url":"https://assets-eu.researchsquare.com/files/rs-9173358/v1/8a53b50585239ade17471b23.mp4"},{"id":107629046,"identity":"5d90487e-4f2f-4a7b-a21e-1c4552a2559e","added_by":"auto","created_at":"2026-04-23 11:19:25","extension":"docx","order_by":4,"title":"","display":"","copyAsset":false,"role":"supplement","size":5935213,"visible":true,"origin":"","legend":"Supplementary document","description":"","filename":"Supplementarydocument.docx","url":"https://assets-eu.researchsquare.com/files/rs-9173358/v1/870a70a11d29bb301146e24e.docx"},{"id":107707547,"identity":"3dfb7168-bd37-4a16-a307-b348be8102fd","added_by":"auto","created_at":"2026-04-24 09:20:32","extension":"mp4","order_by":5,"title":"","display":"","copyAsset":false,"role":"supplement","size":14657982,"visible":true,"origin":"","legend":"Supplemantary Movie 5B","description":"","filename":"SupplementaryMovie5BRHL.mp4","url":"https://assets-eu.researchsquare.com/files/rs-9173358/v1/6e3042bcb049a2a9fd6818d5.mp4"},{"id":107629048,"identity":"2be7d57f-f684-46f3-9dac-08db3a0c833d","added_by":"auto","created_at":"2026-04-23 11:19:25","extension":"mp4","order_by":6,"title":"","display":"","copyAsset":false,"role":"supplement","size":8207740,"visible":true,"origin":"","legend":"Supplemantary Movie 5A","description":"","filename":"SupplementaryMovie5ALHL.mp4","url":"https://assets-eu.researchsquare.com/files/rs-9173358/v1/a8a3e30cda8da987cf20623e.mp4"},{"id":107706360,"identity":"da349e6e-fb91-4b40-a40d-53cd5e3ef2b6","added_by":"auto","created_at":"2026-04-24 09:17:56","extension":"mp4","order_by":7,"title":"","display":"","copyAsset":false,"role":"supplement","size":13661676,"visible":true,"origin":"","legend":"Supplemantary Movie 4A","description":"","filename":"SupplementaryMovie4ALHL.mp4","url":"https://assets-eu.researchsquare.com/files/rs-9173358/v1/61bcca1c92a3c24061c486ef.mp4"},{"id":108006001,"identity":"339e64ac-bbde-4b6f-94bb-965146e3859f","added_by":"auto","created_at":"2026-04-28 12:51:39","extension":"mp4","order_by":8,"title":"","display":"","copyAsset":false,"role":"supplement","size":12434705,"visible":true,"origin":"","legend":"Supplemantary Movie 5C","description":"","filename":"SupplementaryMovie5COfftarget.mp4","url":"https://assets-eu.researchsquare.com/files/rs-9173358/v1/c0b225ff0bd72d9239d5cfd5.mp4"},{"id":107629050,"identity":"b69b23c6-ef15-41a7-85a7-93c5c4a58893","added_by":"auto","created_at":"2026-04-23 11:19:25","extension":"mp4","order_by":9,"title":"","display":"","copyAsset":false,"role":"supplement","size":14719628,"visible":true,"origin":"","legend":"Supplemantary Movie 3B","description":"","filename":"SupplementaryMovie3BToetwitch.mp4","url":"https://assets-eu.researchsquare.com/files/rs-9173358/v1/d1c7a77da58fead7a6a83ee6.mp4"},{"id":108180979,"identity":"f1740e53-bf06-4f61-88c3-6a6a21ba98a7","added_by":"auto","created_at":"2026-04-30 08:55:54","extension":"mp4","order_by":10,"title":"","display":"","copyAsset":false,"role":"supplement","size":15098372,"visible":true,"origin":"","legend":"Supplemantary Movie 4B","description":"","filename":"SupplementaryMovie4BRHL.mp4","url":"https://assets-eu.researchsquare.com/files/rs-9173358/v1/9ab8b5d8edd97bc0578180e7.mp4"}],"financialInterests":"There is \u003cb\u003eNO\u003c/b\u003e Competing Interest.","formattedTitle":"Imaging of Neurovascular Activity during Motor Responses Evoked by Displacement-guided Focused Ultrasound (DgFUS) Neuromodulation in Mice","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"nature-portfolio","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"","title":"Nature Portfolio","twitterHandle":"","acdcEnabled":false,"dfaEnabled":false,"editorialSystem":"ejp","reportingPortfolio":"","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Displacement-guided focused ultrasound (DgFUS), Functional ultrasound (fUS) imaging, Neurovascular response, Cerebral blood volume (CBV), Electromyography (EMG)","lastPublishedDoi":"10.21203/rs.3.rs-9173358/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-9173358/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Focused ultrasound (FUS) shows great promise for precise and non-invasive neuromodulation, with applications from basic neuroscience investigation to the treatment of neurological disorders. Yet, mechanistic monitoring of FUS is challenging and a causal link between FUS-induced displacement and both neurovascular and behavioral responses evoked by FUS in mice remains poorly understood. Here we harness displacement-guided focused ultrasound (DgFUS) to induce target-specific and non-auditory motor responses in anesthetized mice, and functional ultrasound (fUS) to image neurovascular activity during the evoked responses. We show that DgFUS elicits ipsilateral neurovascular responses, in which the primary (i.e. focal-onset) activation localizes with brain displacement, followed by mild contralateral responses. We demonstrate that switching the target hemisphere of DgFUS within the thalamus results in lateralized hindlimb responses concurrent with lateralized neurovascular activation. We show strong correlation between displacement and both cerebral blood volume (CBV) and electromyography (EMG) measurement (Disp. vs. CBV: Spearman r = 0.9091, p \u003c 0.0001, Disp. vs. EMG: r = 0.7332, p = 0.0091, CBV vs. EMG: r = 0.9041, p = 0.0002), indicating displacement-driven mechanism by which DgFUS evokes neurovascular and motor responses. Lastly, we demonstrate anxiety-related behavioral improvement with transcranial fUS-guided FUS delivered to the bilateral hippocampus, consistent with increases in resting-state hippocampal connectivity assessed by transcranial fUS imaging. The findings presented herein will help refine our understanding of neurovascular responses to FUS and the consequential motor and cognitive outcomes.","manuscriptTitle":"Imaging of Neurovascular Activity during Motor Responses Evoked by Displacement-guided Focused Ultrasound (DgFUS) Neuromodulation in Mice","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-04-23 11:18:55","doi":"10.21203/rs.3.rs-9173358/v1","editorialEvents":[],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"communications-engineering","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"commseng","sideBox":"Learn more about [Communications Engineering](http://link.springer.com/journal/44172)","snPcode":"44172","submissionUrl":"https://mts-commseng.nature.com/cgi-bin/main.plex","title":"Communications Engineering","twitterHandle":"@commseng","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"ejp","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"2b28403d-7f82-4ecc-a054-528126f7f373","owner":[],"postedDate":"April 23rd, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[{"id":65156101,"name":"Biological sciences/Neuroscience"},{"id":65156102,"name":"Biological sciences/Biological techniques/Imaging/Ultrasound"}],"tags":[],"updatedAt":"2026-04-23T11:18:55+00:00","versionOfRecord":[],"versionCreatedAt":"2026-04-23 11:18:55","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-9173358","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-9173358","identity":"rs-9173358","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}