Time-restricted feeding promotes skeletal muscle function in diet- and genetic-induced obesity through shared and unique pathways

Time-restricted feeding promotes skeletal muscle function in diet- and genetic-induced obesity through shared and unique pathways | 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 Time-restricted feeding promotes skeletal muscle function in diet- and genetic-induced obesity through shared and unique pathways Christopher Livelo, Yiming Guo, Shweta Varshney, Farah Abou Daya, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-828135/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 21 Feb, 2023 Read the published version in Nature Communications → Version 1 posted You are reading this latest preprint version Abstract Obesity caused by genetic predisposition, a lifestyle of calorie-dense diets and/or circadian disruption can result in complications including metabolic syndrome, cardiovascular disease, and compromised muscle function. By employing time-restricted feeding (TRF), where daily feeding was limited to 12 hours during the day, we observed improved skeletal muscle function compared to ad libitum feeding (ALF). This was observed in both diet-induced obesity (DIO) and genetic-induced obesity (GIO) in a Drosophila melanogaster (fruit fly) model. We evaluated the mechanistic basis of TRF-mediated benefits by utilizing muscle transcriptomic data of indirect flight muscle (IFM) followed by genetic validations, cytological and biochemical evidences. Significant upregulation of glycine N methyltransferase (Gnmt), sarcosine dehydrogenase (Sardh), CG5955 and downregulation of diacylglycerol o-acyltransferase 2 (Dgat2) were commonly induced by TRF intervention under both obese conditions. Moreover, genetic inhibition of Gnmt, Sardh and CG5955 leads to skeletal muscle dysfunction, aberrant lipid accumulation and loss of TRF-mediated benefits. However, skeletal muscle-specific knockdown (KD) of Dgat2 retained muscle function during aging, a result that mimics TRF-mediated benefits. Furthermore, de novo purine biosynthesis appeared to be upregulated specifically in the DIO model under TRF which led to increased ATP levels resulting in improved muscle performance. Additionally, genes associated with AMP kinase (AMPK) signaling, glycogen metabolism, glycolysis, tricarboxylic acid (TCA) cycle and electron transport chain (ETC) signaling were specifically upregulated in GIO model under TRF. TRF mediated benefits in GIO via activation of AMPK, which led to increased ATP levels. Altogether, we identify the shared and distinct pathways in the regulation of muscle function under TRF, which may aid further research and alternative therapeutic avenues that focus on combating comorbidities linked with obesity. Nutrition & Dietetics Physiology Endocrinology & Metabolism Time-restricted feeding diet-induced obesity genetic-obesity circadian rhythm insulin resistance and sensitivity Drosophila genetics skeletal muscle physiology metabolic homeostasis triglycerides obesogenic challenges purine synthesis AMPK-signaling TCA OxPhos ETC glycolysis glycogen metabolism Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Full Text Additional Declarations There is NO Competing Interest. Supplementary Files SupplementaryMaterialsTRFMuscleTranscriptomicsManuscript.pdf This PDF file includes-Supplementary Figures 1-8 SupplementaryTables18TRFMuscleTranscriptomicsManuscript.xlsx This PDF file includes-Supplementary Tables 1-8 Cite Share Download PDF Status: Published Journal Publication published 21 Feb, 2023 Read the published version in Nature Communications → 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-828135","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":50710202,"identity":"9a661d63-900d-4a27-854e-94ec843266be","order_by":0,"name":"Christopher Livelo","email":"","orcid":"","institution":"University of Alabama Birmingham School of Medicine","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Christopher","middleName":"","lastName":"Livelo","suffix":""},{"id":50710203,"identity":"73f40fa7-9b3b-463e-9215-1700ef938077","order_by":1,"name":"Yiming Guo","email":"","orcid":"","institution":"University of California Riverside","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Yiming","middleName":"","lastName":"Guo","suffix":""},{"id":50710204,"identity":"6d7fb462-a5eb-481c-8726-63428a62239d","order_by":2,"name":"Shweta Varshney","email":"","orcid":"","institution":"Salk Institute for Biological Studies","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Shweta","middleName":"","lastName":"Varshney","suffix":""},{"id":50710205,"identity":"a9912d87-6d9e-4413-937c-f3542a3d6c48","order_by":3,"name":"Farah Abou Daya","email":"","orcid":"","institution":"University of Alabama Birmingham School of Medicine","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Farah","middleName":"Abou","lastName":"Daya","suffix":""},{"id":50710206,"identity":"f9b735c5-fc11-4200-b59a-3bcc55bfc30d","order_by":4,"name":"Hiep Le","email":"","orcid":"","institution":"Salk Institute for Biological Studies","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Hiep","middleName":"","lastName":"Le","suffix":""},{"id":50710207,"identity":"790dd98c-3cc1-4c58-8b1c-16e9fdffb0d5","order_by":5,"name":"Satchidananda Panda","email":"","orcid":"","institution":"The Salk Institute for Biological Studies","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Satchidananda","middleName":"","lastName":"Panda","suffix":""},{"id":50710208,"identity":"035b4ef9-b286-4c9f-97e5-969b25230a88","order_by":6,"name":"Girish Melkani","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAxklEQVRIiWNgGAWjYHACxoMNDAw8/AwMbEAOM3F6wFokG0jVwmBwgFgt8mGHDxycUXNHxvhG8rMHDBXWiQ2EtBjeTks4uOHYMx6zG2nmBgxn0onQMjvH4OADtsNALQlmEoxth4nRkv/h4IN/h3mMZ6R/k2D8R4QWeekchoMb2w7zGEjkAG1pIEKLgXSawcGZfYd5JM68KTdIOJZuTNiW2ckPH/Z8O2zP356+7cGHGmtZwrYcQOYlEFIOtoWgoaNgFIyCUTAKADbARVRSaIasAAAAAElFTkSuQmCC","orcid":"https://orcid.org/0000-0003-0353-4633","institution":"University of Alabama Birmingham School of Medicine","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Girish","middleName":"","lastName":"Melkani","suffix":""}],"badges":[],"createdAt":"2021-08-19 20:35:37","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-828135/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-828135/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1038/s41467-023-36474-4","type":"published","date":"2023-02-21T05:00:00+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":13192669,"identity":"668b373b-1be4-413e-ab52-e9add98a19c9","added_by":"auto","created_at":"2021-09-08 18:48:38","extension":"tif","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":646848,"visible":true,"origin":"","legend":"Common differentially expressed genes identified under TRF versus ALF in WT and\nobesity models. (a) Schematic depicting the timing of food access in ALF (ad libitum feeding) and TRF (time-restricted\nfeeding). In ALF, flies have unrestricted access to food. In TRF, flies could consume food only during the\n12 h of daytime.\n(b) Flow chart depicting the experimental set up. Wild type and genetic obese mutant Sk2 flies were\nseparated into respective food conditions regular diet (RD) or high fat diet (HFD) to establish WT, GIO\nand DIO models. Fly models were further separated into either ALF or TRF. Thoraces were collected at\n3 weeks of age every 4 hours totaling to 6 time points for RNA sequencing.\n(c) Venn diagram of significantly upregulated genes performed using DeSeq2 (p value ≤ 0.05, Fold\nchange ≥ 1.25) under TRF in WT, DIO and GIO models.\n(d) Venn diagram of significantly downregulated genes using DeSeq2 (p value ≤0.05, Fold change ≥\n1.25) under TRF in WT, DIO and GIO models.\n(e-g) Volcano plots showing expression profiles of common DEGs under TRF versus ALF in WT (e), DIO\n(f) and GIO (g) models. Red dots represent differentially expressed genes under TRF (p value ≤ 0.05,\nFold change ≥ 1.25).","description":"","filename":"Figure1.tif","url":"https://assets-eu.researchsquare.com/files/rs-828135/v1/fccc390d5ad8f99adfaaeb7b.tif"},{"id":13192667,"identity":"d197c9ac-ba55-4497-b36c-bfcc25eb799f","added_by":"auto","created_at":"2021-09-08 18:48:38","extension":"tif","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":5160975,"visible":true,"origin":"","legend":"Gene expression and functional validation of Gnmt, Sardh and CG5955 in skeletal\nmuscle in WT and obesity models.\n(a) Schematic representation of Gnmt, Sardh and CG5955 connection with glycine utilization and\nproduction with metabolite intermediates and important genes (italic bold) encoding enzymes. Up arrows indicate upregulation of gene expression (fold change ≥ 1.25) and color indicates condition (grey WT,\nOrange DIO, purple GIO). Asterisk indicates p value ≤ 0.05 and fold change ≥ 1.25.\n(b) Expression level (normalized read count, log2) of gene Gnmt, Sardh and CG5955 under ALF and\nTRF in WT, DIO, GIO models.\n(c) Flight performance of WT female flies with Gnmt, Sardh or CG5955 IFM-specific KD at 1 week, 3\nweek and 5 weeks of age. Three independent RNAi lines per gene were tested and plotted individually.\nN# =100-170 females for each independent fly line per time point (N# = 335-528 combined).\n(d) Flight performance of KD in common upregulated genes at 3 weeks of age unchanged in TRF\ncondition. As the three independent lines showed similar results in (c), one fly line from an independent\nbatch was used and N# = 50-60 females for each condition. Bar graphs represent mean and SEM, with\nsignificance presented as nsp \u003e 0.05, *p ≤ 0.05, **p \u003c 0.01, ***p \u003c 0.001.\n(e) Fluorescence images of the IFM from 3-week-old females of control and IFM-specific KD of Gnmt,\nSardh and CG5955 flies upon probing with phalloidin (green) and Nile Red (red puncta). More\naccumulation of lipid (arrows) and actin-containing myofibrillar disorganization was detected in IFM of KD\nof Gnmt, Sardh and CG5955 (asterisks), compared with age-matched control. Scale bar is 20 μm.\n(f-g) Intramuscular lipid quantification (size (f) and density (g)) showed significant increase upon IFMspecific\nKD of Gnmt, Sardh and CG5955 compare to age-matched control. N# = 9 from three flies’ IFM\nper genotype.","description":"","filename":"Figure2.tif","url":"https://assets-eu.researchsquare.com/files/rs-828135/v1/5cbd1b28ae3f5ca73f52e1d1.tif"},{"id":13192876,"identity":"6fb18552-9ca7-43fa-9f38-83ca6a0514fd","added_by":"auto","created_at":"2021-09-08 18:51:38","extension":"tif","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":2728643,"visible":true,"origin":"","legend":"Gene expression and functional validation of Dgat2 in skeletal muscle in WT and\nobesity models.\n(a) Schematic representation of de novo triacylglycerol synthesis with metabolite intermediates and\nimportant genes (italic bold) encoding enzymes. Down arrows indicate downregulation of gene\nexpression and color indicates condition (grey WT, Orange DIO, purple GIO). Asterisk indicates p-value\n\u003c0.05 and fold change \u003e1.25.\n(b) Expression level (normalized read count, log2) of gene Dgat2 under ALF and TRF in WT, DIO, GIO\nmodels.\n(c) Flight performance of WT female flies with Dgat2 KD at 1, 3, 5 and 7 weeks of age. N# = 65-88 female\nflies per time point. Bar graphs represent mean and SEM, with significance presented as nsp \u003e 0.05,\n*p ≤ 0.05, **p \u003c 0.01, ***p \u003c 0.001. (d) Fluorescence images of the IFM from 3-week-old females with IFM-specific KD of Dgat2 upon probing\nwith phalloidin (green) and Nile Red (red puncta). Smaller lipid (arrows) was detected in IFM of KD of\nDgat2 (asterisks), compared with age-matched control. Scale bar is 20 μm.\n(e-f) Intramuscular lipid quantification (size (e) and density (f)) showed significant reduction in lipid droplet\narea upon IFM-specific KD of Dgat2 compare to age-matched control. N# = 9 from three flies’ IFM per\ngenotype.","description":"","filename":"Figure3.tif","url":"https://assets-eu.researchsquare.com/files/rs-828135/v1/077653268b1e77f25e3561e1.tif"},{"id":13192671,"identity":"491595bd-5e7d-448e-9978-cff2cd7fb0ff","added_by":"auto","created_at":"2021-09-08 18:48:38","extension":"tif","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":760650,"visible":true,"origin":"","legend":"TRF upregulated genes linked with purine biosynthesis and folate cycle in DIO model.\n(a) GO analysis and (b) Reactome pathway analysis of 270 genes that were significantly upregulated\nunder TRF in the DIO model. Bar charts represent the -log10 (p value) of each enriched GO term and\npathway. Related pathways are colored similarly. The number of genes identified in each GO term and\npathway is shown in parenthesis.\n(c) Schematic representation of de novo purine biosynthesis and folate cycle with metabolic intermediates\nand important genes (italic bold) encoding enzymes possibly involved in energy regulation in muscle. Up\narrows indicate upregulation of gene expression (fold change ≥ 1.25), asterisk indicates p value ≤ 0.05\nand color indicates condition (grey WT, Orange DIO, purple GIO).\n(d) Expression level (normalized read count, log2) of significantly upregulated genes in de novo purine\nbiosynthesis and folate cycle under TRF versus ALF in the DIO model.\n(e) Heatmap representation of the genes in (d) with periodicity in the DIO model. Asterisk indicates the\ngain of periodicity under TRF.\n(f) Flight performance of control, Nmdmc KD and AdSL KD at 1 and 3 weeks of age. FL indicates\nflightless. N# = 50-70 per condition. FL indicates flightless. Error bars represent SEM. Bar graphs show\nmean and SEM, with significance presented as nsp \u003e 0.05, *p ≤ 0.05, **p \u003c 0.01, ***p \u003c 0.001.\n(g) Flight performance of control, Gnmt KD and Nmdmc KD at 3 weeks of age under HFD ALF, and\nHFD+FA ALF. Error bars represent SEM. N# 40-92 per condition. Bar graphs show mean and SEM, with\nsignificance presented as nsp \u003e 0.05, *p ≤ 0.05, **p \u003c 0.01, ***p \u003c 0.001.\n(h) Relative ATP level measurement in control, IFM-specific KD of Gnmt and Nmdmc at 3 weeks of age\nunder HFD ALF and HFD+FA ALF conditions with N# = 3-5 per condition. Bar graphs represent mean\nand SEM, with significance presented as nsp \u003e 0.05, *p ≤ 0.05, **p \u003c 0.01, ***p \u003c 0.001.","description":"","filename":"Figure4.tif","url":"https://assets-eu.researchsquare.com/files/rs-828135/v1/0b10584a26f9e1030ab9cee6.tif"},{"id":13192672,"identity":"a9944580-2173-472c-9b05-9eaddea17224","added_by":"auto","created_at":"2021-09-08 18:48:38","extension":"tif","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":1120196,"visible":true,"origin":"","legend":"TRF upregulated genes associated with AMPK and downstream pathways in the GIO\nmodel.\n(a) GO analysis and (b) Reactome pathway analysis of 408 genes that were significantly upregulated\nunder TRF in the GIO model. Bar charts represent the -log10 (p value) of each enriched GO term and\npathway. Related pathways are colored similarly. The number of genes identified in each GO term and\npathway is shown in parenthesis.\n(c) Schematic representation of connection between Gnmt, Ampk and downstream of AMPK signaling\nwith important genes (italic bold) encoding enzymes. Up arrows indicate upregulation of gene expression\n(fold change ≥ 1.25) and color indicates condition (grey WT, Orange DIO, purple GIO). Asterisk indicates\np value ≤ 0.05 and fold change ≥ 1.25. The up arrow of Ampk represents the expression of AMPK α\nsubunit.\n(d) Expression level (normalized read count, log2) of significantly upregulated genes in glycolysis,\nglycogen metabolism, TCA cycle and electron transport chain under TRF versus ALF in the GIO model.\n(e) Heatmap representation of the genes in (d) with periodicity in the GIO model. Asterisk indicates the\ngain of periodicity under TRF.\n(f) Representative western blot of p-AMPKα levels (top), AMPKα levels (middle) and α-tubulin (bottom),\nfrom 3-week-old female fly IFMs in WT, DIO and GIO models under ALF (A) and TRF (T).\n(g) Ratios of p-AMPKα/AMPKα normalized to WT ALF with N# = 3.\n(h) Flight index of GIO genes in relation to downstream pathways shown at 1 and 3 weeks of age. N# =\n50-100 per condition. FL indicates flightless. Bar graphs represent mean and SEM, with significance\npresented as nsp \u003e 0.05, *p ≤ 0.05, **p \u003c 0.01, ***p \u003c 0.001.","description":"","filename":"Figure5.tif","url":"https://assets-eu.researchsquare.com/files/rs-828135/v1/8cd01885b4b06002af6ac689.tif"},{"id":13192878,"identity":"5e789104-95d8-43f2-b045-31db1fcadbc2","added_by":"auto","created_at":"2021-09-08 18:51:38","extension":"tif","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":326572,"visible":true,"origin":"","legend":"Proposed mechanism of TRF in Drosophila skeletal muscle under obesogenic\nchallenges.\n12h eating and 12h fasting TRF intervention led to increased SAM regulation by upregulating of Gnmt,\nSardh and CG5955, along with decreased TAG synthesis, which is supported by downregulation of\nDgat2 in all conditions. In the diet-induced obesity model, TRF promoted purine biosynthesis and folate\npool regulation, potentially resulting in higher ATP levels. However, in the genetic-induced obesity\nmodel, AMPK signaling, and downstream pathways were exclusively upregulated via increased p-\nAMPKα levels and increased ATP levels. All pathways regulated led to improved muscle performance\nin obesity models.","description":"","filename":"Figure6.tif","url":"https://assets-eu.researchsquare.com/files/rs-828135/v1/3cb6ea3f73d1b9152695eda2.tif"},{"id":13677287,"identity":"2ea208f6-2c77-4ae3-b418-1745e516bd12","added_by":"auto","created_at":"2021-09-17 11:34:30","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3224676,"visible":true,"origin":"","legend":"","description":"","filename":"TRFMuscleTranscriptomicsManuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-828135/v1_covered.pdf"},{"id":13192879,"identity":"bcb5042e-2357-4e27-a6ba-0f78adffedc5","added_by":"auto","created_at":"2021-09-08 18:51:53","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3221146,"visible":true,"origin":"","legend":"","description":"","filename":"TRFMuscleTranscriptomicsManuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-828135/v1_covered.pdf"},{"id":13192875,"identity":"09baa1b0-6078-460c-ae19-892023bc99ad","added_by":"auto","created_at":"2021-09-08 18:51:38","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":2295404,"visible":true,"origin":"","legend":"This PDF file includes-Supplementary Figures 1-8","description":"","filename":"SupplementaryMaterialsTRFMuscleTranscriptomicsManuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-828135/v1/8507d2aad0b92520b025cede.pdf"},{"id":13192877,"identity":"a568786d-d927-45d7-8400-26bdb9d0f67d","added_by":"auto","created_at":"2021-09-08 18:51:38","extension":"xlsx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":9077802,"visible":true,"origin":"","legend":"This PDF file includes-Supplementary Tables 1-8","description":"","filename":"SupplementaryTables18TRFMuscleTranscriptomicsManuscript.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-828135/v1/0af704e46f83dea08c527f8f.xlsx"}],"financialInterests":"There is \u003cb\u003eNO\u003c/b\u003e Competing Interest.","formattedTitle":"Time-restricted feeding promotes skeletal muscle function in diet- and genetic-induced obesity through shared and unique pathways","fulltext":[{"header":"Full Text","content":"This preprint is available for \u003ca href='/article/rs-828135/latest.pdf' target='_blank'\u003edownload as a PDF\u003c/a\u003e."}],"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":false,"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":"Time-restricted feeding, diet-induced obesity, genetic-obesity, circadian rhythm, insulin resistance and sensitivity, Drosophila genetics, skeletal muscle physiology, metabolic homeostasis, triglycerides, obesogenic challenges, purine synthesis, AMPK-signaling, TCA, OxPhos, ETC, glycolysis, glycogen metabolism","lastPublishedDoi":"10.21203/rs.3.rs-828135/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-828135/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Obesity caused by genetic predisposition, a lifestyle of calorie-dense diets and/or circadian disruption can result in complications including metabolic syndrome, cardiovascular disease, and compromised muscle function. By employing time-restricted feeding (TRF), where daily feeding was limited to 12 hours during the day, we observed improved skeletal muscle function compared to ad libitum feeding (ALF). This was observed in both diet-induced obesity (DIO) and genetic-induced obesity (GIO) in a Drosophila melanogaster (fruit fly) model. We evaluated the mechanistic basis of TRF-mediated benefits by utilizing muscle transcriptomic data of indirect flight muscle (IFM) followed by genetic validations, cytological and biochemical evidences. Significant upregulation of glycine N methyltransferase (Gnmt), sarcosine dehydrogenase (Sardh), CG5955 and downregulation of diacylglycerol o-acyltransferase 2 (Dgat2) were commonly induced by TRF intervention under both obese conditions. Moreover, genetic inhibition of Gnmt, Sardh and CG5955 leads to skeletal muscle dysfunction, aberrant lipid accumulation and loss of TRF-mediated benefits. However, skeletal muscle-specific knockdown (KD) of Dgat2 retained muscle function during aging, a result that mimics TRF-mediated benefits. Furthermore, de novo purine biosynthesis appeared to be upregulated specifically in the DIO model under TRF which led to increased ATP levels resulting in improved muscle performance. Additionally, genes associated with AMP kinase (AMPK) signaling, glycogen metabolism, glycolysis, tricarboxylic acid (TCA) cycle and electron transport chain (ETC) signaling were specifically upregulated in GIO model under TRF. TRF mediated benefits in GIO via activation of AMPK, which led to increased ATP levels. Altogether, we identify the shared and distinct pathways in the regulation of muscle function under TRF, which may aid further research and alternative therapeutic avenues that focus on combating comorbidities linked with obesity.","manuscriptTitle":"Time-restricted feeding promotes skeletal muscle function in diet- and genetic-induced obesity through shared and unique pathways","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2021-09-08 18:48:36","doi":"10.21203/rs.3.rs-828135/v1","editorialEvents":[],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"nature-communications","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"NCOMMS","sideBox":"Learn more about [Nature Communications](http://www.nature.com/ncomms/)","snPcode":"","submissionUrl":"https://mts-ncomms.nature.com/","title":"Nature Communications","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"ejp","reportingPortfolio":"Nature Communications","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"d43639d3-a68c-4838-99fc-402e32beaf9f","owner":[],"postedDate":"September 8th, 2021","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":7057385,"name":"Nutrition \u0026 Dietetics"},{"id":7057386,"name":"Physiology"},{"id":7057387,"name":"Endocrinology \u0026 Metabolism"}],"tags":[],"updatedAt":"2023-02-22T08:06:36+00:00","versionOfRecord":{"articleIdentity":"rs-828135","link":"https://doi.org/10.1038/s41467-023-36474-4","journal":{"identity":"nature-communications","isVorOnly":false,"title":"Nature Communications"},"publishedOn":"2023-02-21 05:00:00","publishedOnDateReadable":"February 21st, 2023"},"versionCreatedAt":"2021-09-08 18:48:36","video":"","vorDoi":"10.1038/s41467-023-36474-4","vorDoiUrl":"https://doi.org/10.1038/s41467-023-36474-4","workflowStages":[]},"version":"v1","identity":"rs-828135","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-828135","identity":"rs-828135","version":["v1"]},"buildId":"7rjqhiLT3MXkJMwkYKINL","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}