Resting-State Theta and Alpha Oscillations in Amputation and Phantom Limb Pain: A Pre-Registered High-Density EEG Study

Resting-State Theta and Alpha Oscillations in Amputation and Phantom Limb Pain: A Pre-Registered High-Density EEG Study | 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 Research Article Resting-State Theta and Alpha Oscillations in Amputation and Phantom Limb Pain: A Pre-Registered High-Density EEG Study Malin Ramne, Eva Lendaro This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8645281/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 7 You are reading this latest preprint version Abstract Phantom limb pain (PLP) affects a substantial portion of individuals with limb amputation, yet its neural mechanisms remain poorly understood. While neuroimaging studies have predominantly employed functional magnetic resonance imaging and magnetoencephalography, resting-state EEG could offer complementary insight into intrinsic brain oscillations but remains largely unexplored in this specific context. More generally, chronic and neuropathic pain have been associated with increased theta-band (4–8 Hz) power and reduced peak alpha frequency (8–13 Hz), but whether these patterns extend to PLP is unknown. Here, we conducted a pre-registered cross-sectional investigation of high-density resting-state EEG (58 channels) in 19 intact controls, 6 amputees without PLP, and 13 amputees with PLP. We employed mixed-effects models with bootstrap inference, multiple sensitivity analyses, and exploratory cluster-based permutation testing. Across primary and sensitivity analyses, we found no evidence for a robust association between the presence of PLP and either theta power or peak alpha frequency. Amputation-related differences in the alpha band reached statistical significance in some analyses, suggesting possible alpha-band alterations associated with limb loss rather than pain per se. An exploratory analysis within the PLP group showed a positive association between one measure of peak alpha frequency and pain intensity. However, inconsistent replication across spectral measures and statistical approaches indicates that these amputation and pain intensity-related findings should be interpreted with caution and require confirmation in larger, well-powered studies. Together, these results suggest that resting-state EEG markers commonly reported in chronic pain do not straightforwardly generalize to PLP. Phantom limb pain Amputation Resting-state EEG electroencephalography BIDS theta alpha oscillations Peak alpha frequency chronic pain Thalamocortical dysrhythmia Alpha slowing Full Text Additional Declarations No competing interests reported. Supplementary Files Supplementalmaterial.docx Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 10 Mar, 2026 Reviews received at journal 26 Feb, 2026 Reviewers agreed at journal 22 Feb, 2026 Reviewers invited by journal 05 Feb, 2026 Editor assigned by journal 02 Feb, 2026 Submission checks completed at journal 02 Feb, 2026 First submitted to journal 20 Jan, 2026 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. 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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-8645281","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":587476166,"identity":"388de81d-ee36-4ebc-936d-7eddbb7c49d9","order_by":0,"name":"Malin Ramne","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA1UlEQVRIiWNgGAWjYLCCBwYMcgwSDIwHiNeSYMBgDNTCQIoWBobEBqK1mLefTvyQUHAnvX9284NDNyru2TPwH8avVeZM7maJBINnuTPuHDM4nHOmGGhdWgJeLRIMuRuAWg5DybaEBAYJHgP8Wvjfbv4BVJxuIJH+4XDuvwSgw85/wK9FIncbyPwEA4kcoC0NCYwNDDl4dQC1vN1mAdRiOONGTsHhnGMJiW0SaYQclrv5xoc/h+X5Z6RvfJxTk2DPz3/4AX5rMAAbiepHwSgYBaNgFGABAO5CShSYtKxvAAAAAElFTkSuQmCC","orcid":"","institution":"Chalmers University of Technology","correspondingAuthor":true,"prefix":"","firstName":"Malin","middleName":"","lastName":"Ramne","suffix":""},{"id":587476167,"identity":"0a7e17e5-a624-445e-b456-0082a0f40b71","order_by":1,"name":"Eva Lendaro","email":"","orcid":"","institution":"Massachusetts Institute of Technology","correspondingAuthor":false,"prefix":"","firstName":"Eva","middleName":"","lastName":"Lendaro","suffix":""}],"badges":[],"createdAt":"2026-01-20 05:40:59","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8645281/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8645281/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":102242363,"identity":"43f53538-a1b8-4d91-bc31-88c551d21f3a","added_by":"auto","created_at":"2026-02-09 17:11:49","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":603985,"visible":true,"origin":"","legend":"","description":"","filename":"Manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8645281/v1_covered_7ce3df85-4132-4a46-aba8-9718c907a559.pdf"},{"id":102242336,"identity":"ef1107be-7cdb-4b6c-80dd-39a30a571926","added_by":"auto","created_at":"2026-02-09 17:11:43","extension":"docx","order_by":0,"title":"","display":"","copyAsset":false,"role":"supplement","size":955716,"visible":true,"origin":"","legend":"","description":"","filename":"Supplementalmaterial.docx","url":"https://assets-eu.researchsquare.com/files/rs-8645281/v1/61bd3a86e9dedb917f748741.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"\u003cp\u003eResting-State Theta and Alpha Oscillations in Amputation and Phantom Limb Pain: A Pre-Registered High-Density EEG Study\u003c/p\u003e","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"brain-topography","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"btop","sideBox":"Learn more about [Brain Topography](http://link.springer.com/journal/10548)","snPcode":"10548","submissionUrl":"https://submission.nature.com/new-submission/10548/3","title":"Brain Topography","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Phantom limb pain, Amputation, Resting-state, EEG, electroencephalography, BIDS, theta, alpha oscillations, Peak alpha frequency, chronic pain, Thalamocortical dysrhythmia, Alpha slowing","lastPublishedDoi":"10.21203/rs.3.rs-8645281/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8645281/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003ePhantom limb pain (PLP) affects a substantial portion of individuals with limb amputation, yet its neural mechanisms remain poorly understood. While neuroimaging studies have predominantly employed functional magnetic resonance imaging and magnetoencephalography, resting-state EEG could offer complementary insight into intrinsic brain oscillations but remains largely unexplored in this specific context. More generally, chronic and neuropathic pain have been associated with increased theta-band (4\u0026ndash;8 Hz) power and reduced peak alpha frequency (8\u0026ndash;13 Hz), but whether these patterns extend to PLP is unknown. Here, we conducted a pre-registered cross-sectional investigation of high-density resting-state EEG (58 channels) in 19 intact controls, 6 amputees without PLP, and 13 amputees with PLP. We employed mixed-effects models with bootstrap inference, multiple sensitivity analyses, and exploratory cluster-based permutation testing. Across primary and sensitivity analyses, we found no evidence for a robust association between the presence of PLP and either theta power or peak alpha frequency. Amputation-related differences in the alpha band reached statistical significance in some analyses, suggesting possible alpha-band alterations associated with limb loss rather than pain per se. An exploratory analysis within the PLP group showed a positive association between one measure of peak alpha frequency and pain intensity. However, inconsistent replication across spectral measures and statistical approaches indicates that these amputation and pain intensity-related findings should be interpreted with caution and require confirmation in larger, well-powered studies. Together, these results suggest that resting-state EEG markers commonly reported in chronic pain do not straightforwardly generalize to PLP.\u003c/p\u003e","manuscriptTitle":"Resting-State Theta and Alpha Oscillations in Amputation and Phantom Limb Pain: A Pre-Registered High-Density EEG Study","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-02-09 17:10:53","doi":"10.21203/rs.3.rs-8645281/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-03-10T16:37:49+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-02-26T19:15:51+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"149628798914536887575442947298874837297","date":"2026-02-23T01:25:27+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2026-02-05T14:38:59+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2026-02-02T07:34:52+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2026-02-02T06:23:50+00:00","index":"","fulltext":""},{"type":"submitted","content":"Brain Topography","date":"2026-01-20T05:29:57+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"brain-topography","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"btop","sideBox":"Learn more about [Brain Topography](http://link.springer.com/journal/10548)","snPcode":"10548","submissionUrl":"https://submission.nature.com/new-submission/10548/3","title":"Brain Topography","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"14694d6b-f778-46ad-9ce6-b19a51d3da05","owner":[],"postedDate":"February 9th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2026-05-01T12:08:05+00:00","versionOfRecord":[],"versionCreatedAt":"2026-02-09 17:10:53","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8645281","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8645281","identity":"rs-8645281","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}