{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T20:03:44Z","timestamp":1768507424105,"version":"3.49.0"},"posted":{"date-parts":[[2023,10,5]]},"group-title":"Neuroscience","reference-count":135,"publisher":"openRxiv","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2023,10,5]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Temporal lobe epilepsy (TLE) is a type of focal epilepsy characterized by spontaneous recurrent seizures originating from the hippocampus. The epigenetic reprogramming hypothesis of epileptogenesis suggests that the development of TLE is associated with alterations in gene transcription changes resulting in a hyperexcitable network in TLE. DNA 5-methylcytosine (5-mC) is an epigenetic mechanism that has been associated with chronic epilepsy. However, the contribution of 5-hydroxymethylcytosine (5-hmC), a product of 5-mC demethylation by the Ten-Eleven Translocation (TET) family proteins in chronic TLE is poorly understood. 5-hmC is abundant in the brain and acts as a stable epigenetic mark altering gene expression through several mechanisms. Here, we found that the levels of bulk DNA 5-hmC but not 5-mC were significantly reduced in the hippocampus of human TLE patients and in the kainic acid (KA) TLE rat model. Using 5-hmC hMeDIP-sequencing, we characterized 5-hmC distribution across the genome and found bidirectional regulation of 5-hmC at intergenic regions within gene bodies. We found that hypohydroxymethylated 5-hmC intergenic regions were associated with several epilepsy-related genes, including\n                  <jats:italic>Gal<\/jats:italic>\n                  ,\n                  <jats:italic>SV2,<\/jats:italic>\n                  and\n                  <jats:italic>Kcnj11<\/jats:italic>\n                  and hyperdroxymethylation 5-hmC intergenic regions were associated with\n                  <jats:italic>Gad65<\/jats:italic>\n                  ,\n                  <jats:italic>TLR4<\/jats:italic>\n                  , and\n                  <jats:italic>Bdnf<\/jats:italic>\n                  gene expression. Mechanistically,\n                  <jats:italic>Tet1<\/jats:italic>\n                  knockdown in the hippocampus was sufficient to decrease 5-hmC levels and increase seizure susceptibility following KA administration. In contrast,\n                  <jats:italic>Tet1<\/jats:italic>\n                  overexpression in the hippocampus resulted in increased 5-hmC levels associated with improved seizure resiliency in response to KA. These findings suggest an important role for 5-hmC as an epigenetic regulator of epilepsy that can be manipulated to influence seizure outcomes.\n                <\/jats:p>","DOI":"10.1101\/2023.10.03.560698","type":"posted-content","created":{"date-parts":[[2023,10,5]],"date-time":"2023-10-05T17:45:18Z","timestamp":1696527918000},"source":"Crossref","is-referenced-by-count":1,"title":["Alterations in DNA 5-hydroxymethylation Patterns in the Hippocampus of an Experimental Model of Refractory Epilepsy"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3748-2766","authenticated-orcid":false,"given":"Rudhab","family":"Bahabry","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8771-3895","authenticated-orcid":false,"given":"Rebecca M.","family":"Hauser","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6409-7786","authenticated-orcid":false,"given":"Richard G.","family":"S\u00e1nchez","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7997-8105","authenticated-orcid":false,"given":"Silvienne Sint","family":"Jago","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1859-2118","authenticated-orcid":false,"given":"Lara","family":"Ianov","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Remy J.","family":"Stuckey","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"R. Ryley","family":"Parrish","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1026-550X","authenticated-orcid":false,"given":"Lawrence Ver","family":"Hoef","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0234-6956","authenticated-orcid":false,"given":"Farah 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