{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,15]],"date-time":"2026-06-15T10:55:46Z","timestamp":1781520946753,"version":"3.54.1"},"reference-count":44,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2024,5,11]],"date-time":"2024-05-11T00:00:00Z","timestamp":1715385600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000062","name":"National Institute of Diabetes and Digestive and Kidney Diseases","doi-asserted-by":"publisher","award":["DK124269"],"award-info":[{"award-number":["DK124269"]}],"id":[{"id":"10.13039\/100000062","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000062","name":"National Institute of Diabetes and Digestive and Kidney Diseases","doi-asserted-by":"publisher","award":["G12MD007585-23"],"award-info":[{"award-number":["G12MD007585-23"]}],"id":[{"id":"10.13039\/100000062","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["DK124269"],"award-info":[{"award-number":["DK124269"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["G12MD007585-23"],"award-info":[{"award-number":["G12MD007585-23"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Type 1 diabetes (T1D) affects gastrointestinal (GI) motility, favoring gastroparesis, constipation, and fecal incontinence, which are more prevalent in women. The mechanisms are unknown. Given the G-protein-coupled estrogen receptor\u2019s (GPER) role in GI motility, we investigated sex-related diabetes-induced epigenetic changes in GPER. We assessed GPER mRNA and protein expression levels using qPCR and Western blot analyses, and quantified the changes in nuclear DNA methyltransferases and histone modifications (H3K4me3, H3Ac, and H3K27Ac) by ELISA kits. Targeted bisulfite and chromatin immunoprecipitation assays were used to evaluate DNA methylation and histone modifications around the GPER promoter by chromatin immunoprecipitation assays in gastric and colonic smooth muscle tissues of male and female control (CTR) and non-obese diabetic (NOD) mice. GPER expression was downregulated in NOD, with sex-dependent variations. In the gastric smooth muscle, not in colonic smooth muscle, downregulation coincided with differences in methylation ratios between regions 1 and 2 of the GPER promoter of NOD. DNA methylation was higher in NOD male colonic smooth muscle than in NOD females. H3K4me3 and H3ac enrichment decreased in NOD gastric smooth muscle. H3K4me3 levels diminished in the colonic smooth muscle of NOD. H3K27ac levels were unaffected, but enrichment decreased in NOD male gastric smooth muscle; however, it increased in the NOD male colonic smooth muscle and decreased in the female NOD colonic smooth muscle. Male NOD colonic smooth muscle exhibited decreased H3K27ac levels, not female, whereas female NOD colonic smooth muscle demonstrated diminished enrichment of H3ac at the GPER promoter, contrary to male NOD. Sex-specific epigenetic mechanisms contribute to T1D-mediated suppression of GPER expression in the GI tract. These insights advance our understanding of T1D complications and suggest promising avenues for targeted therapeutic interventions.<\/jats:p>","DOI":"10.3390\/ijms25105260","type":"journal-article","created":{"date-parts":[[2024,5,14]],"date-time":"2024-05-14T08:03:08Z","timestamp":1715673788000},"page":"5260","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Epigenetic Modulation of GPER Expression in Gastric and Colonic Smooth Muscle of Male and Female Non-Obese Diabetic (NOD) Mice: Insights into H3K4me3 and H3K27ac Modifications"],"prefix":"10.3390","volume":"25","author":[{"given":"Juanita C.","family":"Hixon","sequence":"first","affiliation":[{"name":"Center for Cancer Research, Department of Biology, Tuskegee University, Tuskegee, AL 36088, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jatna I.","family":"Rivas Zarete","sequence":"additional","affiliation":[{"name":"Department of Biomedical Sciences, College of Veterinary Medicine, Tuskegee University, Tuskegee, AL 36088, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jason","family":"White","sequence":"additional","affiliation":[{"name":"Center for Cancer Research, Department of Biology, Tuskegee University, Tuskegee, AL 36088, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mariline","family":"Hilaire","sequence":"additional","affiliation":[{"name":"Department of Environment & Nutrition Sciences, College of Agriculture, Tuskegee University, Tuskegee, AL 36088, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4167-7793","authenticated-orcid":false,"given":"Aliyu","family":"Muhammad","sequence":"additional","affiliation":[{"name":"Center for Cancer Research, Department of Biology, Tuskegee University, Tuskegee, AL 36088, USA"},{"name":"Department of Biochemistry, Faculty of Life Sciences, Ahmadu Bello University, P.M.B. 1044, Zaria 810211, Kaduna State, Nigeria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0404-5233","authenticated-orcid":false,"given":"Abdurrahman Pharmacy","family":"Yusuf","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Federal University of Technology, P.M.B. 65, Minna 920101, Niger State, Nigeria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Benjamin","family":"Adu-Addai","sequence":"additional","affiliation":[{"name":"Department of Biomedical Sciences, College of Veterinary Medicine, Tuskegee University, Tuskegee, AL 36088, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5420-9852","authenticated-orcid":false,"given":"Clayton C.","family":"Yates","sequence":"additional","affiliation":[{"name":"Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins School of Medicine, Johns Hopkins University, Baltimore, MD 21218, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sunila","family":"Mahavadi","sequence":"additional","affiliation":[{"name":"Center for Cancer Research, Department of Biology, Tuskegee University, Tuskegee, AL 36088, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,5,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"14790","DOI":"10.1038\/s41598-020-71908-9","article-title":"Global, regional, and national burden and trend of diabetes in 195 countries and territories: An analysis from 1990 to 2025","volume":"10","author":"Lin","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"109119","DOI":"10.1016\/j.diabres.2021.109119","article-title":"IDF Diabetes Atlas: Global, regional and country-level diabetes prevalence estimates for 2021 and projections for 2045","volume":"183","author":"Sun","year":"2022","journal-title":"Diabetes Res. Clin. Pract."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Gomber, A., Ward, Z.J., Ross, C., Owais, M., Mita, C., Yeh, J.M., Reddy, C.L., and Atun, R. (2022). Variation in the incidence of type 1 diabetes mellitus in children and adolescents by world region and country income group: A scoping review. PLOS Glob. Public Health, 2.","DOI":"10.1371\/journal.pgph.0001099"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"51","DOI":"10.4239\/wjd.v4.i3.51","article-title":"Gastrointestinal complications of diabetes mellitus","volume":"4","author":"Krishnan","year":"2013","journal-title":"World J. Diabetes"},{"key":"ref_5","first-page":"263","article-title":"Diabetic Gastroenteropathy: A Complication of Diabetes Mellitus","volume":"51","author":"Kurniawan","year":"2019","journal-title":"Acta Med. Indones."},{"key":"ref_6","first-page":"1697","article-title":"Gastrointestinal complications of diabetes","volume":"77","author":"Shakil","year":"2008","journal-title":"Am. Fam. Physician"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"794","DOI":"10.4239\/wjd.v12.i6.794","article-title":"Diabetic gastroenteropathy: An underdiagnosed complication","volume":"12","year":"2021","journal-title":"World J. Diabetes"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"213","DOI":"10.5056\/jnm15186","article-title":"Calcium Sensitization Mechanisms in Gastrointestinal Smooth Muscles","volume":"22","author":"Perrino","year":"2016","journal-title":"J. Neurogastroenterol. Motil."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"407","DOI":"10.1038\/s41574-023-00822-7","article-title":"The G protein-coupled oestrogen receptor GPER in health and disease: An update","volume":"19","author":"Prossnitz","year":"2023","journal-title":"Nat. Rev. Endocrinol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"715","DOI":"10.1038\/nrendo.2011.122","article-title":"The G-protein-coupled estrogen receptor GPER in health and disease","volume":"7","author":"Prossnitz","year":"2011","journal-title":"Nat. Rev. Endocrinol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"100080","DOI":"10.1016\/j.endmts.2021.100080","article-title":"Targeting the G protein-coupled estrogen receptor (GPER) in obesity and diabetes","volume":"2","author":"Sharma","year":"2021","journal-title":"Endocr. Metab. Sci."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"427","DOI":"10.1007\/978-3-319-70178-3_20","article-title":"G-Protein-Coupled Estrogen Receptor (GPER) and Sex-Specific Metabolic Homeostasis","volume":"1043","author":"Sharma","year":"2017","journal-title":"Adv. Exp. Med. Biol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1016\/j.jsbmb.2017.02.012","article-title":"Roles of G protein-coupled estrogen receptor GPER in metabolic regulation","volume":"176","author":"Sharma","year":"2018","journal-title":"J. Steroid Biochem. Mol. Biol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"627","DOI":"10.2337\/dc17-1536","article-title":"Gastrointestinal Symptoms in Diabetes: Prevalence, Assessment, Pathogenesis, and Management","volume":"41","author":"Du","year":"2018","journal-title":"Diabetes Care"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"14015","DOI":"10.1038\/s41598-023-41112-6","article-title":"Prevalence of gastroparesis in diabetic patients: A systematic review and meta-analysis","volume":"13","author":"Li","year":"2023","journal-title":"Sci. Rep."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Li, Z.L., Liu, J.C., Liu, S.B., Li, X.Q., Yi, D.H., and Zhao, M.G. (2012). Improvement of vascular function by acute and chronic treatment with the GPR30 agonist G1 in experimental diabetes mellitus. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0038787"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"e13025","DOI":"10.1111\/nmo.13025","article-title":"G protein-coupled estrogen receptor and estrogen receptor ligands regulate colonic motility and visceral pain","volume":"29","author":"Fichna","year":"2017","journal-title":"Neurogastroenterol. Motil. Off. J. Eur. Gastrointest. Motil. Soc."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2145","DOI":"10.1161\/CIRCULATIONAHA.110.956839","article-title":"Epigenetic modifications: Basic mechanisms and role in cardiovascular disease","volume":"123","author":"Handy","year":"2011","journal-title":"Circulation"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1141","DOI":"10.1080\/15592294.2019.1638701","article-title":"DNA hypermethylation in disease: Mechanisms and clinical relevance","volume":"14","author":"Ehrlich","year":"2019","journal-title":"Epigenetics"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"15782","DOI":"10.1073\/pnas.0607617103","article-title":"The genomic landscape of histone modifications in human T cells","volume":"103","author":"Roh","year":"2006","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"289","DOI":"10.1007\/978-94-007-4525-4_13","article-title":"The role of DNA methylation and histone modifications in transcriptional regulation in humans","volume":"61","author":"Miller","year":"2013","journal-title":"Subcell. Biochem."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1184","DOI":"10.1111\/ejn.15935","article-title":"Prenatal exposure of diabetes and progestin-mediated autistic biomarker in peripheral blood mononuclear cells","volume":"57","author":"Jiao","year":"2023","journal-title":"Eur. J. Neurosci."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"432","DOI":"10.1111\/nmo.12743","article-title":"G protein-coupled estrogen receptor is involved in modulating colonic motor function via nitric oxide release in C57BL\/6 female mice","volume":"28","author":"Li","year":"2016","journal-title":"Neurogastroenterol. Motil. Off. J. Eur. Gastrointest. Motil. Soc."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1522","DOI":"10.1101\/gr.156539.113","article-title":"Functional DNA methylation differences between tissues, cell types, and across individuals discovered using the M&M algorithm","volume":"23","author":"Zhang","year":"2013","journal-title":"Genome Res."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s13072-015-0047-z","article-title":"Genetic and epigenetic variation among inbred mouse littermates: Identification of inter-individual differentially methylated regions","volume":"8","author":"Oey","year":"2015","journal-title":"Epigenetics Chromatin"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"972","DOI":"10.1101\/gr.101535.109","article-title":"Chromatin and sequence features that define the fine and gross structure of genomic methylation patterns","volume":"20","author":"Edwards","year":"2010","journal-title":"Genome Res."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1038\/npp.2012.112","article-title":"DNA methylation and its basic function","volume":"38","author":"Moore","year":"2013","journal-title":"Neuropsychopharmacol. Off. Publ. Am. Coll. Neuropsychopharmacol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"e324","DOI":"10.1038\/emm.2017.11","article-title":"Writing, erasing and reading histone lysine methylations","volume":"49","author":"Hyun","year":"2017","journal-title":"Exp. Mol. Med."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"138","DOI":"10.1186\/s13148-021-01126-1","article-title":"The dynamic broad epigenetic (H3K4me3, H3K27ac) domain as a mark of essential genes","volume":"13","author":"Beacon","year":"2021","journal-title":"Clin. Epigenetics"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"3668","DOI":"10.1073\/pnas.1813565116","article-title":"Epigenomic analysis reveals DNA motifs regulating histone modifications in human and mouse","volume":"116","author":"Ngo","year":"2019","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"15912","DOI":"10.1038\/s41598-021-95398-5","article-title":"Investigating crosstalk between H3K27 acetylation and H3K4 trimethylation in CRISPR\/dCas-based epigenome editing and gene activation","volume":"11","author":"Zhao","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1038\/s41421-022-00514-y","article-title":"Dynamics of histone acetylation during human early embryogenesis","volume":"9","author":"Wu","year":"2023","journal-title":"Cell Discov."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"5633","DOI":"10.1038\/s41598-024-54213-7","article-title":"Sex-specific epigenetics drive low GPER expression in gastrointestinal smooth muscles in type 2 diabetic mice","volume":"14","author":"Muhammad","year":"2024","journal-title":"Sci. Rep."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1235","DOI":"10.1038\/embor.2009.218","article-title":"Structural basis for recognition of H3K4 methylation status by the DNA methyltransferase 3A ATRX-DNMT3-DNMT3L domain","volume":"10","author":"Otani","year":"2009","journal-title":"EMBO Rep."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1362","DOI":"10.1016\/j.bbagrm.2014.02.007","article-title":"Understanding the relationship between DNA methylation and histone lysine methylation","volume":"1839","author":"Rose","year":"2014","journal-title":"Biochim. Biophys. Acta-Gene Regul. Mech."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"e13495","DOI":"10.1111\/nmo.13495","article-title":"Muscarinic m2 receptor-mediated actin polymerization via PI3 kinase \u03b3 and integrin-linked kinase in gastric smooth muscle","volume":"31","author":"Mahavadi","year":"2019","journal-title":"Neurogastroenterol. Motil."},{"key":"ref_37","unstructured":"(2022, May 04). Bio-Rad. Real-Time PCR Applications Guide (Bulletin 5279 US\/EG Rev B). Available online: https:\/\/www.bio-rad.com\/webroot\/web\/pdf\/lsr\/literature\/Bulletin_5279.pdf."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"mfad042","DOI":"10.1093\/mtomcs\/mfad042","article-title":"Regulation of Atp7a RNA contributes to differentiation-dependent Cu redistribution in skeletal muscle cells","volume":"15","author":"Whitlow","year":"2023","journal-title":"Metallomics"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1217","DOI":"10.1007\/s10695-019-00640-7","article-title":"Optimum salinity for Nile tilapia (Oreochromis niloticus) growth and mRNA transcripts of ion-regulation, inflammatory, stress-and immune-related genes","volume":"45","author":"Hemeda","year":"2019","journal-title":"Fish Physiol. Biochem."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"3114","DOI":"10.1172\/JCI27702","article-title":"TRPC6 fulfills a calcineurin signaling circuit during pathologic cardiac remodeling","volume":"116","author":"Kuwahara","year":"2006","journal-title":"J. Clin. Investig."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1693","DOI":"10.1094\/PDIS-08-18-1387-RE","article-title":"Treatment with wheat root exudates and soil microorganisms from wheat\/watermelon companion cropping can induce watermelon disease resistance against Fusarium oxysporum f. sp. niveum","volume":"103","author":"Li","year":"2019","journal-title":"Plant Dis."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"270","DOI":"10.1039\/c2ib00127f","article-title":"MudPIT analysis of released proteins in Pseudomonas aeruginosa laboratory and clinical strains in relation to pro-inflam-matory effects","volume":"4","author":"Bergamini","year":"2012","journal-title":"Integr. Biol."},{"key":"ref_43","first-page":"1453","article-title":"Ticagrelor suppresses oxidized low-density lipoprotein-induced endothelial cell apoptosis and alleviates atherosclerosis in ApoE\u2212\/\u2212 mice via downregulation of PCSK9","volume":"19","author":"Xia","year":"2019","journal-title":"Mol. Med. Rep."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1998","DOI":"10.1007\/s12035-017-0457-y","article-title":"Differences in 5-HT2A and mGlu2 receptor expression levels and repressive epigenetic modifications at the 5-HT2A promoter region in the Roman low-(RLA-I) and high-(RHA-I) avoidance rat strains","volume":"55","author":"Fomsgaard","year":"2018","journal-title":"Mol. Neurobiol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/10\/5260\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T14:40:56Z","timestamp":1760107256000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/10\/5260"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,5,11]]},"references-count":44,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2024,5]]}},"alternative-id":["ijms25105260"],"URL":"https:\/\/doi.org\/10.3390\/ijms25105260","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,5,11]]}}}