{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T05:10:17Z","timestamp":1788844217017,"version":"build-2803163510"},"reference-count":43,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2026,9,6]],"date-time":"2026-09-06T00:00:00Z","timestamp":1788652800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Good Samaritan Research Foundation"},{"name":"NIH","award":["R01 NS084920"],"award-info":[{"award-number":["R01 NS084920"]}]},{"award":["R01 NS084920"],"award-info":[{"award-number":["R01 NS084920"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Children\u2019s Health Research Institute (CHRI) of UNMC"},{"id":[{"id":"https:\/\/ror.org\/038pa9k74","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Int. J. Mol. Sci.","IJMS"],"abstract":"<jats:p>DNA methylation is implicated in epileptogenesis. Sarcosine, a glycine transporter 1 (GlyT1) inhibitor and methyl donor, attenuates behavioral progression during rapid hippocampal kindling and alters hippocampal DNA methylation, but its locus-specific epigenetic effects remain poorly understood. Here, reduced representation bisulfite sequencing (RRBS) was combined with targeted gene expression analysis in the hippocampi of sarcosine-treated kindled rats. RRBS identified 563, 533, and 390 differentially methylated regions (DMRs), corresponding to 521, 499, and 374 DMR-associated genes, in vehicle-kindled versus sham (vKD vs. vSH), sarcosine-kindled versus sham (sKD vs. vSH), and sarcosine-kindled versus vehicle-kindled (sKD vs. vKD) comparisons, respectively. Pathway enrichment analysis identified 217 significantly affected pathways, including glutamatergic signaling, extracellular matrix (ECM) organization, chromatin regulation, axon guidance, and apoptotic processes. Eleven candidate genes involved in epigenetic regulation, excitatory neurotransmission, and ECM remodeling were selected for transcriptional validation. All 11 genes were significantly upregulated in kindled hippocampi, whereas sarcosine was associated with reduced expression relative to vehicle-kindled rats for eight genes (Hdac9, Fos, Smad7, Unc5a, Grik2, Gpr37l1, Cacna2d2, and Yy1). Collectively, these findings indicate that sarcosine remodels DNA methylation-associated transcriptional networks during rapid hippocampal kindling and support GlyT1 inhibition as a potential disease-modifying approach in experimental epileptogenesis.<\/jats:p>","DOI":"10.3390\/ijms27177938","type":"journal-article","created":{"date-parts":[[2026,9,7]],"date-time":"2026-09-07T13:53:28Z","timestamp":1788789208000},"page":"7938","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Sarcosine Remodels DNA Methylation-Linked Transcriptional Networks During Epileptogenesis in the Rat Rapid Hippocampal Kindling Model"],"prefix":"10.3390","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0009-0004-2304-2366","authenticated-orcid":false,"given":"Nicole","family":"Ferris","sequence":"first","affiliation":[{"name":"Division of Neurology, Department of Pediatrics, University of Nebraska Medical Center, Children\u2019s Nebraska, Omaha, NE 68198, USA"},{"name":"Child Health Research Institute, University of Nebraska Medical Center, Children\u2019s Nebraska, Omaha, NE 68198, USA"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-3891-482X","authenticated-orcid":false,"given":"Lan","family":"Phung","sequence":"additional","affiliation":[{"name":"Department of Translational Neuroscience, Legacy Research Institute, Portland, OR 97232, USA"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"investigation","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"}]},{"given":"Wakaba","family":"Omi","sequence":"additional","affiliation":[{"name":"Department of Translational Neuroscience, Legacy Research Institute, Portland, OR 97232, USA"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"data-curation","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"}]},{"given":"Guoku","family":"Hu","sequence":"additional","affiliation":[{"name":"Department of Pharmacology & Experimental Neuroscience, University of Nebraska Medical Center, Omaha, NE 68198, USA"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"investigation","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8430-2978","authenticated-orcid":false,"given":"Hai-Ying","family":"Shen","sequence":"additional","affiliation":[{"name":"Division of Neurology, Department of Pediatrics, University of Nebraska Medical Center, Children\u2019s Nebraska, Omaha, NE 68198, USA"},{"name":"Child Health Research Institute, University of Nebraska Medical Center, Children\u2019s Nebraska, Omaha, NE 68198, USA"},{"name":"Department of Translational Neuroscience, Legacy Research Institute, Portland, OR 97232, USA"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,6]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"459","DOI":"10.1038\/s41582-019-0217-x","article-title":"Neuroinflammatory pathways as treatment targets and biomarkers in epilepsy","volume":"15","author":"Vezzani","year":"2019","journal-title":"Nat. Rev. Neurol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"108","DOI":"10.1016\/j.pharmthera.2017.03.002","article-title":"Epigenetic interventions for epileptogenesis: A new frontier for curing epilepsy","volume":"177","author":"Younus","year":"2017","journal-title":"Pharmacol. Ther."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Berger, T.C., Taub\u00f8ll, E., and Heuser, K. (2022). The potential role of DNA methylation as preventive treatment target of epileptogenesis. Front. Cell. Neurosci., 16.","DOI":"10.3389\/fncel.2022.931356"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"616","DOI":"10.1093\/brain\/awu373","article-title":"Differential DNA methylation profiles of coding and non-coding genes define hippocampal sclerosis in human temporal lobe epilepsy","volume":"138","author":"Bryan","year":"2015","journal-title":"Brain"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1186\/s13148-019-0793-z","article-title":"Epigenome-wide association study of seizures in childhood and adolescence","volume":"12","author":"Caramaschi","year":"2020","journal-title":"Clin. Epigenet."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"420","DOI":"10.1007\/s12031-011-9602-7","article-title":"Increased expression of DNA methyltransferase 1 and 3a in human temporal lobe epilepsy","volume":"46","author":"Zhu","year":"2012","journal-title":"J. Mol. Neurosci."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2413","DOI":"10.1111\/epi.17351","article-title":"Genome-wide decrease in DNA methylation in adults with epilepsy treated with modified ketogenic diet: A prospective study","volume":"63","author":"Pedersen","year":"2022","journal-title":"Epilepsia"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Tao, H., Chen, Z., Wu, J., Chen, J., Chen, Y., Fu, J., Sun, C., Zhou, H., Zhong, W., and Zhou, X. (2021). DNA Methylation Signature of Epileptic Encephalopathy-Related Pathogenic Genes Encoding Ion Channels in Temporal Lobe Epilepsy. Front. Neurol., 12.","DOI":"10.3389\/fneur.2021.692412"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1047","DOI":"10.2174\/1570159X23666241220163832","article-title":"Epigenetic Mechanisms in the Pathophysiology and Progression of Epilepsy: A Comprehensive Review of Experimental and Clinical Studies","volume":"23","author":"Li","year":"2025","journal-title":"Curr. Neuropharmacol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1577","DOI":"10.1523\/JNEUROSCI.5180-11.2012","article-title":"Differential DNA methylation patterns define status epilepticus and epileptic tolerance","volume":"32","author":"Das","year":"2012","journal-title":"J. Neurosci."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1111\/epi.12031","article-title":"The emerging role of DNA methylation in epileptogenesis","volume":"53","author":"Kobow","year":"2012","journal-title":"Epilepsia"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Berger, T.C., Vigeland, M.D., Hjorthaug, H.S., Etholm, L., Nome, C.G., Taub\u00f8ll, E., Heuser, K., and Selmer, K.K. (2019). Neuronal and glial DNA methylation and gene expression changes in early epileptogenesis. PLoS ONE, 14.","DOI":"10.1371\/journal.pone.0226575"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"186","DOI":"10.1177\/1073858417705840","article-title":"The Epigenetics of Epilepsy and Its Progression","volume":"24","author":"Hauser","year":"2018","journal-title":"Neuroscientist"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"25668","DOI":"10.1038\/srep25668","article-title":"Etiology matters\u2014Genomic DNA Methylation Patterns in Three Rat Models of Acquired Epilepsy","volume":"6","author":"Debski","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"105657","DOI":"10.1016\/j.neuint.2023.105657","article-title":"Epigenetic regulation in epilepsy: A novel mechanism and therapeutic strategy for epilepsy","volume":"173","author":"Chen","year":"2024","journal-title":"Neurochem. Int."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1016\/j.eplepsyres.2018.11.006","article-title":"The molecular hallmarks of epigenetic effects mediated by antiepileptic drugs","volume":"149","author":"Rocha","year":"2019","journal-title":"Epilepsy Res."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"919","DOI":"10.1056\/NEJMra1004418","article-title":"Drug-resistant epilepsy","volume":"365","author":"Kwan","year":"2011","journal-title":"N. Engl. J. Med."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"5095","DOI":"10.1038\/s41598-026-35826-6","article-title":"Commonalities in gene expression and methylation changes across two rat models of acquired epilepsy","volume":"16","author":"Purnell","year":"2026","journal-title":"Sci. Rep."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Shen, H.Y., Weltha, L., Cook, J.M., Gesese, R., Omi, W., Baer, S.B., Rose, R.M., Reemmer, J., and Boison, D. (2020). Sarcosine Suppresses Epileptogenesis in Rats with Effects on Hippocampal DNA Methylation. Front. Mol. Neurosci., 13.","DOI":"10.3389\/fnmol.2020.00097"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1186\/s12929-016-0314-8","article-title":"Effects of sarcosine and N, N-dimethylglycine on NMDA receptor-mediated excitatory field potentials","volume":"24","author":"Lee","year":"2017","journal-title":"J. Biomed. Sci."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"441","DOI":"10.1038\/329441a0","article-title":"Kindling stimulation induces c-fos protein(s) in granule cells of the rat dentate gyrus","volume":"329","author":"Dragunow","year":"1987","journal-title":"Nature"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"192","DOI":"10.1126\/science.3037702","article-title":"Mapping patterns of c-fos expression in the central nervous system after seizure","volume":"237","author":"Morgan","year":"1987","journal-title":"Science"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"8927","DOI":"10.1523\/JNEUROSCI.0430-09.2009","article-title":"Transcriptome profiling reveals TGF-beta signaling involvement in epileptogenesis","volume":"29","author":"Cacheaux","year":"2009","journal-title":"J. Neurosci."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"3209","DOI":"10.1007\/s10787-026-02210-9","article-title":"From insult to hyperexcitability: Pharmacological targeting of MyD88 and JAK\/STAT3 pathways in epilepsy","volume":"34","author":"Abdelaziz","year":"2026","journal-title":"Inflammopharmacology"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Bortoletto, R., Balestrieri, M., Bhattacharyya, S., and Colizzi, M. (2022). Is It Time to Test the Antiseizure Potential of Palmitoylethanolamide in Human Studies? A Systematic Review of Preclinical Evidence. Brain Sci., 12.","DOI":"10.3390\/brainsci12010101"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"a022434","DOI":"10.1101\/cshperspect.a022434","article-title":"Role of astrocytes in epilepsy","volume":"5","author":"Coulter","year":"2015","journal-title":"Cold Spring Harb. Perspect. Med."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"12983","DOI":"10.1007\/s00210-026-05276-y","article-title":"GlyT1 (SLC6A9) inhibition in neurological and psychiatric disorders","volume":"399","author":"Cavalcante","year":"2026","journal-title":"Naunyn-Schmiedeberg\u2019s Arch. Pharmacol."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Auer, S., Hoffmann, L., Schicht, M., Blumcke, I., and Paulsen, F. (2026). Temporal lobe epilepsy is associated with neuroinflammation, extracellular matrix remodeling, and synaptic protein alterations. Front. Mol. Neurosci., 19.","DOI":"10.3389\/fnmol.2026.1728666"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"7369020","DOI":"10.1155\/2016\/7369020","article-title":"Matrix Metalloproteinase 9 in Epilepsy: The Role of Neuroinflammation in Seizure Development","volume":"2016","author":"Bronisz","year":"2016","journal-title":"Mediat. Inflamm."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1111\/epi.13629","article-title":"Perampanel effects in the WAG\/Rij rat model of epileptogenesis, absence epilepsy, and comorbid depressive-like behavior","volume":"58","author":"Citraro","year":"2017","journal-title":"Epilepsia"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"413","DOI":"10.1038\/nm.2127","article-title":"Toll-like receptor 4 and high-mobility group box-1 are involved in ictogenesis and can be targeted to reduce seizures","volume":"16","author":"Maroso","year":"2010","journal-title":"Nat. Med."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1055","DOI":"10.1016\/S1474-4422(06)70626-3","article-title":"Effects of seizures on developmental processes in the immature brain","volume":"5","author":"Holmes","year":"2006","journal-title":"Lancet Neurol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"755","DOI":"10.1016\/S0079-6123(07)63041-6","article-title":"Epileptogenesis in the dentate gyrus: A critical perspective","volume":"163","author":"Dudek","year":"2007","journal-title":"Prog. Brain Res."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"E22","DOI":"10.3171\/FOC\/2008\/25\/9\/E22","article-title":"The bumetanide-sensitive Na-K-2Cl cotransporter NKCC1 as a potential target of a novel mechanism-based treatment strategy for neonatal seizures","volume":"25","author":"Kahle","year":"2008","journal-title":"Neurosurg. Focus"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1404","DOI":"10.1007\/s12035-023-03630-2","article-title":"Knockdown of SIK3 in the CA1 Region can Reduce Seizure Susceptibility in Mice by Inhibiting Decreases in GABAAR \u03b11 Expression","volume":"61","author":"Jiang","year":"2024","journal-title":"Mol. Neurobiol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"e138332","DOI":"10.1172\/JCI138332","article-title":"The matrix metalloproteinase inhibitor IPR-179 has antiseizure and antiepileptogenic effects","volume":"131","author":"Broekaart","year":"2021","journal-title":"J. Clin. Investig."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"273","DOI":"10.1177\/15357597211018688","article-title":"Perineuronal Net Dynamics in the Pathophysiology of Epilepsy","volume":"21","author":"Chaunsali","year":"2021","journal-title":"Epilepsy Curr."},{"key":"ref_38","first-page":"ENEURO-0379","article-title":"Differential Expression and Cell-Type Specificity of Perineuronal Nets in Hippocampus, Medial Entorhinal Cortex, and Visual Cortex Examined in the Rat and Mouse","volume":"4","author":"Christensen","year":"2017","journal-title":"eNeuro"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"741","DOI":"10.1007\/s00401-013-1168-8","article-title":"Deep sequencing reveals increased DNA methylation in chronic rat epilepsy","volume":"126","author":"Kobow","year":"2013","journal-title":"Acta Neuropathol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1210","DOI":"10.1021\/acschemneuro.5c00039","article-title":"Effect of SSR504734, a Selective Glycine Transporter Type 1 Inhibitor, on Seizure Thresholds, Neurotransmitter Levels, and Inflammatory Markers in Mice","volume":"16","author":"Gapinska","year":"2025","journal-title":"ACS Chem. Neurosci."},{"key":"ref_41","unstructured":"Paxinos, G., and Watson, C. (2007). The Rat Brain in Stereotaxic Coordinates, Academic Press. [6th ed.]."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"234","DOI":"10.1227\/00006123-197809000-00018","article-title":"Kindling: The first decade","volume":"3","author":"Racine","year":"1978","journal-title":"Neurosurgery"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"19232","DOI":"10.1038\/s41598-019-55291-8","article-title":"Synergistic Effects of Hyperandrogenemia and Obesogenic Western-Style Diet on Transcription and DNA Methylation in Visceral Adipose Tissue of Nonhuman Primates","volume":"9","author":"Carbone","year":"2019","journal-title":"Sci. Rep."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/17\/7938\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T04:24:55Z","timestamp":1788841495000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/17\/7938"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,6]]},"references-count":43,"journal-issue":{"issue":"17","published-online":{"date-parts":[[2026,9]]}},"alternative-id":["ijms27177938"],"URL":"https:\/\/doi.org\/10.3390\/ijms27177938","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,6]]}}}