{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T13:14:21Z","timestamp":1785417261590,"version":"3.56.0"},"posted":{"date-parts":[[2026,2,9]]},"group-title":"Developmental Biology","reference-count":69,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,2,9]],"date-time":"2026-02-09T00:00:00Z","timestamp":1770595200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"award":["R01EY013760"],"award-info":[{"award-number":["R01EY013760"]}],"id":[{"id":"https:\/\/ror.org\/03wkg3b53","id-type":"ROR","asserted-by":"publisher"}]},{"award":["R21EY033473"],"award-info":[{"award-number":["R21EY033473"]}],"id":[{"id":"https:\/\/ror.org\/03wkg3b53","id-type":"ROR","asserted-by":"publisher"}]},{"award":["P30EY008216"],"award-info":[{"award-number":["P30EY008216"]}],"id":[{"id":"https:\/\/ror.org\/03wkg3b53","id-type":"ROR","asserted-by":"publisher"}]},{"award":["T32EY007135"],"award-info":[{"award-number":["T32EY007135"]}],"id":[{"id":"https:\/\/ror.org\/03wkg3b53","id-type":"ROR","asserted-by":"publisher"}]},{"award":["F31EY0355654"],"award-info":[{"award-number":["F31EY0355654"]}],"id":[{"id":"https:\/\/ror.org\/03wkg3b53","id-type":"ROR","asserted-by":"publisher"}]},{"id":[{"id":"https:\/\/ror.org\/04drjs621","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,2,9]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Mueller glial reprogramming studies demonstrate that mammalian Mueller glia can be induced to proliferate and\/or engage in neural differentiation, as occurs naturally in teleost fish. A major objective is the identification of combined strategies that promote both robust proliferation and neurogenesis. These studies would benefit from a translatable screening platform that enables controlled perturbation, maintained tissue context and longitudinal analysis, such as 3D culture for first tier analysis of reprogramming strategies. Here, we validate a 3D retinal culture for Mueller glial reprogramming studies by recapitulating key signatures of an\n                  <jats:italic>in vivo<\/jats:italic>\n                  reprogramming paradigm. Next, we find that electrical stimulation (E-stim) as a tunable, extrinsic cue is sufficient to activate endogenous Ascl1 expression, indicating a state transition favorable for neurogenesis, while Mueller glia-specific\n                  <jats:italic>\n                    p27\n                    <jats:sup>Kip<\/jats:sup>\n                  <\/jats:italic>\n                  <jats:sup>1<\/jats:sup>\n                  inactivation promotes robust, prolonged proliferation. Utilization of the lineage-tracing proliferation-history reporter H3.1-iCOUNT enabled longitudinal proliferation analysis and assessment of reprogramming outcomes within the proliferative, Mueller-derived population. With this model, we find that E-stim and\n                  <jats:italic>\n                    p27\n                    <jats:sup>Kip<\/jats:sup>\n                  <\/jats:italic>\n                  <jats:sup>1<\/jats:sup>\n                  inactivation in combination (ESPI) increases proliferation, endogenous Ascl1 expression, and neurogenesis of Mueller-derived cells across modalities. Together, this work establishes a 3D culture framework for discovery of combinatorial reprogramming strategies within a proliferative context and identifies ESPI as an efficient approach to proliferative, neurogenic Mueller glial reprogramming.\n                <\/jats:p>","DOI":"10.64898\/2026.02.05.704102","type":"posted-content","created":{"date-parts":[[2026,2,9]],"date-time":"2026-02-09T19:45:17Z","timestamp":1770666317000},"source":"Crossref","is-referenced-by-count":0,"title":["Electrical stimulation combined with p27Kip1 inactivation drives proliferative neurogenic reprogramming of Mueller glia in the adult mouse retina"],"prefix":"10.64898","author":[{"given":"Megan L.","family":"Stone","sequence":"first","affiliation":[{"name":"Department of Cell and Developmental Biology, Vanderbilt University, Nashville, TN, 37232"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joel","family":"Jovanovic","sequence":"additional","affiliation":[{"name":"Vanderbilt Eye Institute, Vanderbilt University Medical Center, Nashville, TN, 37232"},{"name":"Department of Ophthalmology and Visual Sciences, Vanderbilt University, Nashville, TN, 37232"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1725-2805","authenticated-orcid":false,"given":"Edward M.","family":"Levine","sequence":"additional","affiliation":[{"name":"Department of Cell and Developmental Biology, Vanderbilt University, Nashville, TN, 37232"},{"name":"Vanderbilt Eye Institute, Vanderbilt University Medical Center, Nashville, TN, 37232"},{"name":"Department of Ophthalmology and Visual Sciences, Vanderbilt University, Nashville, TN, 37232"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026021108365024000_2026.02.05.704102v1.1","doi-asserted-by":"publisher","DOI":"10.1002\/dneu.20362"},{"key":"2026021108365024000_2026.02.05.704102v1.2","doi-asserted-by":"publisher","DOI":"10.1038\/nrn3723"},{"key":"2026021108365024000_2026.02.05.704102v1.3","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-genet-111523-102000"},{"key":"2026021108365024000_2026.02.05.704102v1.4","doi-asserted-by":"publisher","DOI":"10.3389\/fncel.2023.1305896"},{"key":"2026021108365024000_2026.02.05.704102v1.5","doi-asserted-by":"publisher","DOI":"10.1002\/dneu.20596"},{"key":"2026021108365024000_2026.02.05.704102v1.6","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.adn2091"},{"key":"2026021108365024000_2026.02.05.704102v1.7","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.0938-24.2024"},{"issue":"8","key":"2026021108365024000_2026.02.05.704102v1.8","first-page":"1101","article-title":"Reassessing 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