Subtle changes in crosslinking drive diverse anomalous transport characteristics in actin-microtubule networks

Subtle changes in crosslinking drive diverse anomalous transport characteristics in actin-microtubule networks | 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 Subtle changes in crosslinking drive diverse anomalous transport characteristics in actin-microtubule networks Sylas Anderson, Jonathan Garamella, Ryan McGorty, Rae Robertson-Anderson This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-119120/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Anomalous diffusion in crowded and complex environments is widely studied due to its importance in intracellular transport, fluid rheology and materials engineering. Specifically, diffusion through the cytoskeleton, a network comprised of semiflexible actin filaments and rigid microtubules that interact both sterically and via crosslinking, plays a principal role in viral infection, vesicle transport and targeted drug delivery. Here, we elucidate the impact of crosslinking on particle diffusion in composites of actin and microtubules with actin-actin, microtubule-microtubule and actin-microtubule crosslinking. We analyze a suite of complementary transport metrics by coupling single-particle tracking and differential dynamic microscopy. Using these orthogonal techniques, we find that particles display non-Gaussian and non-ergodic subdiffusion that is markedly enhanced by cytoskeletal crosslinking of any type, which we attribute to suppressed microtubule mobility. However, the extent to which transport deviates from normal Brownian diffusion depends strongly on the crosslinking motif – with actin-microtubule crosslinking inducing the most pronounced anomalous characteristics – due to increased actin fluctuation heterogeneity. Our results reveal that subtle changes to actin-microtubule interactions can have dramatic impacts on diffusion in the cytoskeleton, and suggest that less mobile and more locally heterogeneous networks lead to more strongly anomalous transport. Medical Physics Biophysics Applied & Industrial Microbiology intracellular transport fluid rheology materials engineering cytoskeleton Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Full Text Cite Share Download PDF Status: Posted 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. 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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-119120","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":5642179,"identity":"38e61748-5b44-4e60-80ef-d00834ecf317","order_by":0,"name":"Sylas Anderson","email":"","orcid":"","institution":"University of San Diego","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Sylas","middleName":"","lastName":"Anderson","suffix":""},{"id":5642180,"identity":"dbb20cf6-e527-4445-838e-96f0b80e3d5e","order_by":1,"name":"Jonathan Garamella","email":"","orcid":"","institution":"University of San Diego","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Jonathan","middleName":"","lastName":"Garamella","suffix":""},{"id":5642181,"identity":"6344b800-e9a1-4c12-8ea4-10ec8178a451","order_by":2,"name":"Ryan McGorty","email":"","orcid":"","institution":"University of San Diego","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Ryan","middleName":"","lastName":"McGorty","suffix":""},{"id":5642182,"identity":"85288b70-0899-4b37-b276-da476705e22a","order_by":3,"name":"Rae Robertson-Anderson","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA80lEQVRIiWNgGAWjYJACCTDJ3gAk2CBsBh6itPAcIFmLRAJICwNhLebtZw/e+LinTs5c8vnDzwVlFnLy7g2MD9624dYicyYv2XLGs8PGlrNzjKVnnJMwNjxzgNlwLh4tEgw5ZtI8Bw4kbridwyDN2yaRuHFGAhuQgUcL/xsz6T8H6hI33Dz++DdQSz1QC/tvvFokgLYwHGBO3HCDwQxkS4K8RAIbM34tb4wtew4cNjY4k2NmzXNOwnADz8FmyTnn8Dksx/DGjwN1cgbHjz++zVNWJy/f3nzww5sy3FowgcEBxgZS1AOBPKkaRsEoGAWjYNgDAFBnTjtwN4XsAAAAAElFTkSuQmCC","orcid":"","institution":"University of San Diego","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Rae","middleName":"","lastName":"Robertson-Anderson","suffix":""}],"badges":[],"createdAt":"2020-12-01 00:44:01","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-119120/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-119120/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":3996343,"identity":"a7a46230-7690-4403-83c2-9ddb043cce37","added_by":"auto","created_at":"2020-12-03 18:29:19","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":335398,"visible":true,"origin":"","legend":"Experimental approach to examine the impact of crosslinking on anomalous\ntransport in cytoskeleton networks.","description":"","filename":"Fig1.png","url":"https://assets-eu.researchsquare.com/files/rs-119120/v1/539119ff9643071db857f4e0.png"},{"id":3996344,"identity":"8f23ba02-dac3-426a-9933-7c3d56082af5","added_by":"auto","created_at":"2020-12-03 18:29:19","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":57310,"visible":true,"origin":"","legend":"Crosslinking of cytoskeleton networks leads to multi-phase particle transport with\nmore subdiffusive behavior at long times. ","description":"","filename":"Fig2.png","url":"https://assets-eu.researchsquare.com/files/rs-119120/v1/81be4bf776fb2e68365fd559.png"},{"id":3996345,"identity":"f9653b60-766f-44db-8338-e04506ae7c7d","added_by":"auto","created_at":"2020-12-03 18:29:19","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":67619,"visible":true,"origin":"","legend":"van Hove distributions reveal non-Gaussian, heterogeneous particle transport in\nall networks.","description":"","filename":"Fig3.png","url":"https://assets-eu.researchsquare.com/files/rs-119120/v1/09c444f04dfdba9b4517126d.png"},{"id":3996346,"identity":"3bef3d31-120b-49db-9797-50aed289f8b2","added_by":"auto","created_at":"2020-12-03 18:29:19","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":33147,"visible":true,"origin":"","legend":"DDM analysis reveals heterogenous, non-ergodic transport amplified in\ncrosslinked networks.","description":"","filename":"Fig4.png","url":"https://assets-eu.researchsquare.com/files/rs-119120/v1/b3bfa10b597eadf9e4449c96.png"},{"id":3996347,"identity":"20f7b468-19e0-4f37-86a4-55e05c135124","added_by":"auto","created_at":"2020-12-03 18:29:19","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":41599,"visible":true,"origin":"","legend":"Metrics from both SPT and DDM show that crosslinking increases the non-\nGaussianity and non-ergodicity of particle transport.","description":"","filename":"Fig5.png","url":"https://assets-eu.researchsquare.com/files/rs-119120/v1/a50f5e5865cecfa1829e3695.png"},{"id":3996348,"identity":"0287ce67-bc2a-4fca-8d11-5b34638fef6d","added_by":"auto","created_at":"2020-12-03 18:29:19","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":50322,"visible":true,"origin":"","legend":"Multiple transport metrics highlight the degree to which crosslinking motif drives deviations from normal Brownian motion in composite cytoskeleton networks.","description":"","filename":"Fig6.png","url":"https://assets-eu.researchsquare.com/files/rs-119120/v1/fc23da49dd3262b68d2c90ab.png"},{"id":13562189,"identity":"aad195d6-ae93-4653-b0b3-60f83d3589b1","added_by":"auto","created_at":"2021-09-17 03:12:08","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2140309,"visible":true,"origin":"","legend":"","description":"","filename":"SAndersonSRNov20Main.pdf","url":"https://assets-eu.researchsquare.com/files/rs-119120/v1_covered.pdf"},{"id":3996349,"identity":"3b851dfb-6bfd-4c1d-bad1-c3daedbfc07a","added_by":"auto","created_at":"2020-12-03 18:29:28","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2372219,"visible":true,"origin":"","legend":"","description":"","filename":"SAndersonSRNov20Main.pdf","url":"https://assets-eu.researchsquare.com/files/rs-119120/v1_stamped.pdf"}],"financialInterests":"","formattedTitle":"Subtle changes in crosslinking drive diverse anomalous transport characteristics in actin-microtubule networks","fulltext":[{"header":"Full Text","content":"\u003cp\u003eThis preprint is available for \u003ca href='/article/rs-119120/latest.pdf' target='_blank'\u003edownload as a PDF\u003c/a\u003e.\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"intracellular transport, fluid rheology, materials engineering, cytoskeleton","lastPublishedDoi":"10.21203/rs.3.rs-119120/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-119120/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Anomalous diffusion in crowded and complex environments is widely studied due to its importance in intracellular transport, fluid rheology and materials engineering. Specifically, diffusion through the cytoskeleton, a network comprised of semiflexible actin filaments and rigid microtubules that interact both sterically and via crosslinking, plays a principal role in viral infection, vesicle transport and targeted drug delivery. Here, we elucidate the impact of crosslinking on particle diffusion in composites of actin and microtubules with actin-actin, microtubule-microtubule and actin-microtubule crosslinking. We analyze a suite of complementary transport metrics by coupling single-particle tracking and differential dynamic microscopy. Using these orthogonal techniques, we find that particles display non-Gaussian and non-ergodic subdiffusion that is markedly enhanced by cytoskeletal crosslinking of any type, which we attribute to suppressed microtubule mobility. However, the extent to which transport deviates from normal Brownian diffusion depends strongly on the crosslinking motif – with actin-microtubule crosslinking inducing the most pronounced anomalous characteristics – due to increased actin fluctuation heterogeneity. Our results reveal that subtle changes to actin-microtubule interactions can have dramatic impacts on diffusion in the cytoskeleton, and suggest that less mobile and more locally heterogeneous networks lead to more strongly anomalous transport.","manuscriptTitle":"Subtle changes in crosslinking drive diverse anomalous transport characteristics in actin-microtubule networks","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2020-12-03 18:29:17","doi":"10.21203/rs.3.rs-119120/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"d4e7607f-10da-41d6-9d83-c5534c44d047","owner":[],"postedDate":"December 3rd, 2020","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":1316480,"name":"Medical Physics"},{"id":1316481,"name":"Biophysics"},{"id":1316482,"name":"Applied \u0026 Industrial Microbiology"}],"tags":[],"updatedAt":"2021-01-13T06:29:17+00:00","versionOfRecord":[],"versionCreatedAt":"2020-12-03 18:29:17","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-119120","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-119120","identity":"rs-119120","version":["v1"]},"buildId":"FbvkV6FR0MCFSLy54lSbu","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}