Thermoneutral Environment Improves Mouse Metabolism and Reduces Stress in Metabolic Cages

Thermoneutral Environment Improves Mouse Metabolism and Reduces Stress in Metabolic Cages | 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 Thermoneutral Environment Improves Mouse Metabolism and Reduces Stress in Metabolic Cages Petra Seebeck, Philipp Villiger, Charlotte Calvet, Eva Pastor-Arroyo, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4509623/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 10 Oct, 2025 Read the published version in Lab Animal → Version 1 posted You are reading this latest preprint version Abstract Metabolic cages [MCs] are frequently used to collect feces and urine samples. However, MC housing is stressful for mice and thereby potentially influences parameters of interest. We compared a standard protocol for MC housing (4 days at 23°C, including 3 days of permanent acclimatisation and 24h sampling) with a) short-term intermittent acclimatisation (3 days for 3h plus 24h MC housing), b) providing a nest (4 days at 23°C) and c) MC housing at thermoneutrality (4 days at 30°C). C57BL6/N mice were implanted with telemetric transmitters acquiring ECG, blood pressure, body core temperature, and activity. Body core temperature decreased, and heart and respiratory rates increased during MC housing at 23°C. Mice housed in MCs with a nest or at 30°C showed strongly reduced heart/respiratory rates and a higher body core temperature compared to the group housed at 23°C. Mice ate more and lost more weight when housed in MCs at 23°C in combination with a much higher brown fat activity. Torpor-like episodes were observed in male mice housed at 23°C. In addition, they had higher corticosterone levels. Our study demonstrates that MC housing at 23°C strongly influences mouse physiology and metabolism due to a substantial cold stress. Female mice seem to be less affected compared to male mice. MC housing at thermoneutrality offers a simple solution to improve animal wellbeing during MC housing. Moreover, one short-term acclimatization presumably has the same acclimatisation effect as repeated exposure to the MCs. Biological sciences/Physiology/Metabolism Biological sciences/Physiology/Circulation Biological sciences/Physiology/Respiration Metabolic cage Temperature Stress Laboratory Mice Refinement Distress Full Text Additional Declarations There is NO Competing Interest. The authors declare that the research was conducted without any commercial or financial relationships that could be construed as a potential conflict of interest. CAW reports honoraria from Kyowa Kirin and Medice outside this project. Cite Share Download PDF Status: Published Journal Publication published 10 Oct, 2025 Read the published version in Lab Animal → 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-4509623","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":324737920,"identity":"f43db2fc-0591-4593-b6fd-6bb8ff7a5d1c","order_by":0,"name":"Petra Seebeck","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA5UlEQVRIiWNgGAWjYFADCQYGxoYKBgYDBgY2AkqZkbWcIVlLYxsRWszZ+499+MFQJ8cv3Xzw48x5h+3NGZifPcCnxbLnMPPMHobDxpJzjiVLbtx2mNmygc3cAJ8WgxvJzAw8DAcSN9zIMWN8uO0wm8EBHjYJvFruP2Zm/MNQV7//Rv43xodzDvMQ1nKDmZmZh4E5wUAih41xY8NhCYJaLHuSjZllDA4bzrhzzFhyxrF0A4PDbGZ4tZizH3zM+KaiTp5/dvPDjz011vYGx5uf4XcYEgkFzFgVomsZBaNgFIyCUYAPAAC9Z0OhM+1EiQAAAABJRU5ErkJggg==","orcid":"https://orcid.org/0000-0002-8398-5495","institution":"Zurich Integrative Rodent Physiology (ZIRP), University of Zurich","correspondingAuthor":true,"prefix":"","firstName":"Petra","middleName":"","lastName":"Seebeck","suffix":""},{"id":324737921,"identity":"d8af5bb2-7bc9-47d3-a26d-73dac2408a9a","order_by":1,"name":"Philipp Villiger","email":"","orcid":"https://orcid.org/0000-0001-5286-3332","institution":"Institute of Physiology, University of Zurich and Zurich Integrative Rodent Physiology (ZIRP)","correspondingAuthor":false,"prefix":"","firstName":"Philipp","middleName":"","lastName":"Villiger","suffix":""},{"id":324737922,"identity":"41dac43d-7b9b-40ad-8945-1cdbc87a0f86","order_by":2,"name":"Charlotte Calvet","email":"","orcid":"https://orcid.org/0000-0002-6163-9807","institution":"Institute of Physiology, University of Zurich and Zurich Integrative Rodent Physiology (ZIRP)","correspondingAuthor":false,"prefix":"","firstName":"Charlotte","middleName":"","lastName":"Calvet","suffix":""},{"id":324737923,"identity":"fd97fd09-b6df-4dbd-aeea-84f8808fee10","order_by":3,"name":"Eva Pastor-Arroyo","email":"","orcid":"","institution":"Institute of Physiology, University of Zurich","correspondingAuthor":false,"prefix":"","firstName":"Eva","middleName":"","lastName":"Pastor-Arroyo","suffix":""},{"id":324737924,"identity":"f0eb13c5-4e55-4729-b482-88c776cbbdfe","order_by":4,"name":"Carsten Wagner","email":"","orcid":"https://orcid.org/0000-0002-9874-8898","institution":"University of Zurich","correspondingAuthor":false,"prefix":"","firstName":"Carsten","middleName":"","lastName":"Wagner","suffix":""}],"badges":[],"createdAt":"2024-05-31 14:26:11","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4509623/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4509623/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1038/s41684-025-01618-0","type":"published","date":"2025-10-10T04:00:00+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":93285670,"identity":"da1ed85a-186b-48e8-914a-8c983dab4fed","added_by":"auto","created_at":"2025-10-11 07:09:31","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3443940,"visible":true,"origin":"","legend":"","description":"","filename":"ThermoneutralEnvironmentImprovesMouseMetabolismandReducesStressinMetabolicCages.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4509623/v1_covered_343ea81c-5657-40cf-b72f-bc35ba3886e7.pdf"}],"financialInterests":"There is \u003cb\u003eNO\u003c/b\u003e Competing Interest.\nThe authors declare that the research was conducted without any commercial or financial relationships that could be construed as a potential conflict of interest. CAW reports honoraria from Kyowa Kirin and Medice outside this project.","formattedTitle":"Thermoneutral Environment Improves Mouse Metabolism and Reduces Stress in Metabolic Cages","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"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":"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":"Metabolic cage, Temperature Stress, Laboratory Mice, Refinement, Distress","lastPublishedDoi":"10.21203/rs.3.rs-4509623/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4509623/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eMetabolic cages [MCs] are frequently used to collect feces and urine samples. However, MC housing is stressful for mice and thereby potentially influences parameters of interest.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eWe compared a standard protocol for MC housing (4 days at 23°C, including 3 days of permanent acclimatisation and 24h sampling) with a) short-term intermittent acclimatisation (3 days for 3h plus 24h MC housing), b) providing a nest (4 days at 23°C) and c) MC housing at thermoneutrality (4 days at 30°C). C57BL6/N mice were implanted with telemetric transmitters acquiring ECG, blood pressure, body core temperature, and activity.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eBody core temperature decreased, and heart and respiratory rates increased during MC housing at 23°C. Mice housed in MCs with a nest or at 30°C showed strongly reduced heart/respiratory rates and a higher body core temperature compared to the group housed at 23°C. Mice ate more and lost more weight when housed in MCs at 23°C in combination with a much higher brown fat activity. Torpor-like episodes were observed in male mice housed at 23°C. In addition, they had higher corticosterone levels.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eOur study demonstrates that MC housing at 23°C strongly influences mouse physiology and metabolism due to a substantial cold stress. Female mice seem to be less affected compared to male mice. MC housing at thermoneutrality offers a simple solution to improve animal wellbeing during MC housing. Moreover, one short-term acclimatization presumably has the same acclimatisation effect as repeated exposure to the MCs.\u0026nbsp;\u003c/p\u003e","manuscriptTitle":"Thermoneutral Environment Improves Mouse Metabolism and Reduces Stress in Metabolic Cages","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-07-11 02:19:19","doi":"10.21203/rs.3.rs-4509623/v1","editorialEvents":[],"status":"published","journal":{"display":false,"email":"[email protected]","identity":"lab-animal","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"laban","sideBox":"Learn more about [Lab Animal](http://www.nature.com/laban/)","snPcode":"","submissionUrl":"","title":"Lab Animal","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"ejp","reportingPortfolio":"Nature Research","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"beabc9ba-f3b2-4582-86a0-4346a8f345e1","owner":[],"postedDate":"July 11th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":34348733,"name":"Biological sciences/Physiology/Metabolism"},{"id":34348734,"name":"Biological sciences/Physiology/Circulation"},{"id":34348735,"name":"Biological sciences/Physiology/Respiration"}],"tags":[],"updatedAt":"2025-10-11T07:09:20+00:00","versionOfRecord":{"articleIdentity":"rs-4509623","link":"https://doi.org/10.1038/s41684-025-01618-0","journal":{"identity":"lab-animal","isVorOnly":false,"title":"Lab Animal"},"publishedOn":"2025-10-10 04:00:00","publishedOnDateReadable":"October 10th, 2025"},"versionCreatedAt":"2024-07-11 02:19:19","video":"","vorDoi":"10.1038/s41684-025-01618-0","vorDoiUrl":"https://doi.org/10.1038/s41684-025-01618-0","workflowStages":[]},"version":"v1","identity":"rs-4509623","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4509623","identity":"rs-4509623","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}