Projecting neurons from lateral entorhinal cortex to basolateral amygdala mediate the encoding of incidental odor-taste associations

Projecting neurons from lateral entorhinal cortex to basolateral amygdala mediate the encoding of incidental odor-taste associations | 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 Projecting neurons from lateral entorhinal cortex to basolateral amygdala mediate the encoding of incidental odor-taste associations Arnau Busquets-Garcia, Jose Antonio González-Parra, Vittoria Acciai, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5754062/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 Daily choices are determined by prior direct or indirect associations between low-salience cues and reinforcers. In this study, we used a mouse odor-taste sensory preconditioning task combined with genetic, intersectional and chemogenetic approaches to identify a novel brain circuit involved in mediated learning. We found that neuronal projections from the lateral entorhinal cortex to the basolateral amygdala are engaged during low-salience stimuli associations, which is essential for mediated learning formation. Biological sciences/Neuroscience/Neural circuits Biological sciences/Neuroscience/Emotion/Amygdala Figures Figure 1 Figure 2 Figure 3 Full Text Additional Declarations There is NO Competing Interest. Supplementary Files GonzXXlezParraetalSupplementaryInformation.pdf Supplementary Material 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-5754062","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":398974060,"identity":"56c0617c-e372-49d7-bc62-8c84a5971bcb","order_by":0,"name":"Arnau Busquets-Garcia","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABCklEQVRIiWNgGAWjYDACCRiDvQFdhIAWCQaeA2haeAhqkUggUgv/7B7jD2/bbOrkZ759+PBHhV0+/+zuxM8VDHfk7HFZcueMmeTctjQJg9vpxsY8Z5ItZ9w5u1nyDMMzY5wOu5FjxszbdljCQDqNTZqxjdnAQCJ3g2QDw+HEHhw65G/kGH/mbfsvIT/zGPvPn//qQVo2/wRqqcelxeBGjoE0b9sBCYYbbGwMvA2HQVq2gWxJwOUwwzvHyiTnnEuW3HAmjVma59hxA4kbudssGwwOG/YcwK5F7nbz5g9vyuz45duPMX78UVNtwD8jd/PNhorD8vC4xQawOMEAj3IcWkbBKBgFo2AUIAAADORXrxHNxkcAAAAASUVORK5CYII=","orcid":"https://orcid.org/0000-0002-7100-8873","institution":"Hospital del Mar Research Institute","correspondingAuthor":true,"prefix":"","firstName":"Arnau","middleName":"","lastName":"Busquets-Garcia","suffix":""},{"id":398974061,"identity":"54abed7d-d71f-4968-bf46-6df1a02c2ede","order_by":1,"name":"Jose Antonio González-Parra","email":"","orcid":"","institution":"Hospital del Mar Research Institute","correspondingAuthor":false,"prefix":"","firstName":"Jose","middleName":"Antonio","lastName":"González-Parra","suffix":""},{"id":398974062,"identity":"75c5db5f-e5a7-4eff-9a7c-44d6afcc5ad4","order_by":2,"name":"Vittoria Acciai","email":"","orcid":"","institution":"Hospital del Mar Research Institute","correspondingAuthor":false,"prefix":"","firstName":"Vittoria","middleName":"","lastName":"Acciai","suffix":""},{"id":398974063,"identity":"c39aef61-53a3-4e78-9c58-0f0478c1c2b5","order_by":3,"name":"Laura Vidal-Palencia","email":"","orcid":"","institution":"Hospital del Mar Research Institute","correspondingAuthor":false,"prefix":"","firstName":"Laura","middleName":"","lastName":"Vidal-Palencia","suffix":""},{"id":398974064,"identity":"311f485f-c7ba-432f-b37e-00593e00d4c7","order_by":4,"name":"Marc Canela-Grimau","email":"","orcid":"","institution":"Hospital del Mar Research Institute","correspondingAuthor":false,"prefix":"","firstName":"Marc","middleName":"","lastName":"Canela-Grimau","suffix":""}],"badges":[],"createdAt":"2025-01-02 21:15:09","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-5754062/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5754062/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":78825487,"identity":"01a59a87-1890-4c05-8539-a48ea9953b78","added_by":"auto","created_at":"2025-03-19 12:23:42","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":1427369,"visible":true,"origin":"","legend":"\u003cp\u003eEnhanced Activity in the Basolateral Amygdala (BLA) During Odor-Taste Incidental Associations. (A) Schematic representation of the odor-taste sensory preconditioning paradigm. Percentage of liquid consumption during mediated (B) and direct (C) learning tests in C57BL/6J mice. (D) Schematic of the protocol used in TRAP2:Ai14 mice to investigate brain regions activated during incidental associations (preconditioning phase) and/or retrieval of mediated learning. (E) Percentage of liquid consumption during mediated learning test in TRAP2:Ai14 mice. Quantification of tdTomato-positive cells during the preconditioning phase (F), c-Fos-positive cells during mediated learning test (G) and colocalized tdTomato/c-Fos-positive cells (H) in the BLA of TRAP2:Ai14 mice. Data are represented as mean values and before-after for liquid consumption or as mean ± SEM for image-based quantifications. For statistical details and n, see Supplementary Table 1. Scale Bar: 25 μm. *** p\u0026lt;0.001; * p\u0026lt;0.05. mCS (mediated conditioned stimulus); CS (conditioned stimulus).\u003c/p\u003e","description":"","filename":"Binder11.png","url":"https://assets-eu.researchsquare.com/files/rs-5754062/v1/c149666046d4d538db453d17.png"},{"id":78824465,"identity":"2f6ea542-05df-465f-9eaf-f3243977d5de","added_by":"auto","created_at":"2025-03-19 12:15:42","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":904645,"visible":true,"origin":"","legend":"\u003cp\u003eBLA Is Required for Encoding Incidental Associations during odor-taste sensory preconditioning. (A) Schematic illustration of BLA inhibition during the preconditioning phase in C57BL/6J mice. (B) Representative image showing DREADDGi expression in the BLA. Percentage of liquid consumption during mediated learning test (C) and direct learning test (D) in C57BL/6J mice. (E) Schematic procedure to inhibit activated preconditioning BLA neurons during mediated learning test in TRAP2:Ai14 mice. (F) Representative image of mCherry expression in the BLA. Percentage of liquid consumption during mediated learning test (G) and direct learning test (H) in TRAP2:Ai14 mice. Data are represented as mean values and before-after for liquid consumption. For statistical details and n, see Supplementary Table 1. Scale Bar: 25 μm. *** p\u0026lt;0.001; * p\u0026lt;0.05. mCS (mediated conditioned stimulus); CS (conditioned stimulus)\u003c/p\u003e","description":"","filename":"Binder12.png","url":"https://assets-eu.researchsquare.com/files/rs-5754062/v1/dda47e4fe8c1c19ba622894a.png"},{"id":78824470,"identity":"6e2e52b8-fdf9-4aae-89ce-93fbef19a19d","added_by":"auto","created_at":"2025-03-19 12:15:43","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":947688,"visible":true,"origin":"","legend":"\u003cp\u003eProjections from LEnt to BLA are Critical for the encoding of incidental associations. (A) Schematic procedure to identify activated brain regions projecting to the BLA during preconditioning. (B) Percentage of liquid consumption during mediated learning test. (C) Percentage of activated cells during the preconditioning phase that also project to the BLA (%tdTomato+GFP/tdTomato+) and (D) representative image of the LEnt. (F) Experimental protocol to inactivated LEnt-BLA projection during incidental associations. (G) Representative image showing DREADD-Gi expression in the LEnt and retro-GFP expression in the BLA. Percentage of liquid consumption during mediated learning test (H) and direct learning test (I) in C57BL/6J mice. Data are represented as mean values and before-after for liquid consumption or as mean ± SEM for image-based quantifications. For statistical details and n, see Supplementary Table 1. Scale Bar: 25 μm. White arrows indicate vTdtomato+GFP+ (colocalization) cells*** p\u0026lt;0.001.OFC: Orbitofrontal cortex; Pir: Piriform cortex; PrH: Perirhinal cortex; vCA1: ventral CA1; LEnt: Lateral Entorrinhal cortex. mCS (mediated conditioned stimulus); CS (conditioned stimulus)\u003c/p\u003e","description":"","filename":"Binder13.png","url":"https://assets-eu.researchsquare.com/files/rs-5754062/v1/c47c7f57d2ba7bc9f3741bcc.png"},{"id":78826568,"identity":"4bd75061-3c0b-4536-bbec-fdbadf1a048c","added_by":"auto","created_at":"2025-03-19 12:39:48","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1140018,"visible":true,"origin":"","legend":"Article File","description":"","filename":"GonzXXlezParraetalMainText.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5754062/v1_covered_5c15e31a-f89d-4c0e-ad26-14f1630b406c.pdf"},{"id":78824472,"identity":"41448786-30c6-4577-8416-41ee6b9df895","added_by":"auto","created_at":"2025-03-19 12:15:43","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":3771897,"visible":true,"origin":"","legend":"Supplementary Material","description":"","filename":"GonzXXlezParraetalSupplementaryInformation.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5754062/v1/8f650c756d00db396e91aabc.pdf"}],"financialInterests":"There is \u003cb\u003eNO\u003c/b\u003e Competing Interest.","formattedTitle":"Projecting neurons from lateral entorhinal cortex to basolateral amygdala mediate the encoding of incidental odor-taste associations","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"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":"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":"","lastPublishedDoi":"10.21203/rs.3.rs-5754062/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5754062/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Daily choices are determined by prior direct or indirect associations between low-salience cues and reinforcers. In this study, we used a mouse odor-taste sensory preconditioning task combined with genetic, intersectional and chemogenetic approaches to identify a novel brain circuit involved in mediated learning. 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