Nitroreductase-mediated cell ablation uncovers the pivotal role of ependymoglial cells in entire cortex regeneration in axolotls

Nitroreductase-mediated cell ablation uncovers the pivotal role of ependymoglial cells in entire cortex regeneration in axolotls | 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 Nitroreductase-mediated cell ablation uncovers the pivotal role of ependymoglial cells in entire cortex regeneration in axolotls Ji-Feng Fei, Sulei Fu, Yan-Yun Zeng, Cheng Peng, Liqun Wang, Yuxian Feng, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6585454/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 20 Jan, 2026 Read the published version in Nature Communications → Version 1 posted You are reading this latest preprint version Abstract Developing precise targeted cell ablation in the axolotl (Ambystoma mexicanum) is crucial for elucidating the roles or interactions of specific cell types in regeneration and modeling diseases. Here we establish a Nitroreductase (NTR)-based inducible cell ablation system in axolotls. Through generation of Sox2:Cherry-NTR knock-in axolotls, we achieve efficient ablation of ependymoglial cells (EGCs) in the central nervous system. Combined spinal cord and brain transplantation and injury models demonstrate regeneration failure upon EGC depletion, suggesting that EGCs are solo source of central nervous system regeneration. Additionally, EGC ablation in the spinal cord resulted in delayed tail regeneration. Moreover, we establish NeuroD6:Cherry-NTR and NeuroD6:Cherry-NTR2.0 knock-in lines to ablate postmitotic cortical neurons, to model degenerative disease and study the process of brain regeneration after large scale neuronal depletion. We found that NTR2.0 (but not NTR) leads to elimination of >95% of cortical neurons. Following the depletion of nearly all cortical neurons, all lost neuronal subtypes are chronological regenerated with layer organization mirroring developmental patterning. Finally, we create Cre-LoxP-based conditional NTR2.0 transgenic axolotls using a constitutive CAGGs promoter, enabling tissue-specific ablation of the targeted cell when combine established Cre lines. In summary, our study establishes an efficient and versatile targeted cell ablation system in axolotls, providing a valuable tool for deep dissection of tissue regeneration in axolotls. Biological sciences/Stem cells/Regeneration Biological sciences/Biological techniques Axolotl Cell ablation Cortex NeuroD6 NTR2.0 Sox2 Full Text Additional Declarations There is NO Competing Interest. Supplementary Files Fuetal.SupplementrayInformation.pdf Supplementray Information SupplementaryVideo1Neurod6NTR2.0MTZ10dpt5xspeed.mp4 Supplementary Video 1 Neurod6:CherryNTR2.0 predatory behavior at 10-day post MTZ treatment.mp4 SupplementaryVideo2Neurod6NTR2.0DMSO10dpt5xspeed.mp4 Supplementary Video 2 Neurod6:CherryNTR2.0 predatory behavior at 10-day post DMSO treatment.mp4 SupplementaryVideo3Neurod6NTR2.0MTZ50dpt5xspeed.mp4 Supplementary Video 3 Neurod6:CherryNTR2.0 predatory behavior at 50-day post MTZ treatment.mp4 SupplementaryVideo4Neurod6NTR2.0DMSO50dpt5xspeed.mp4 Supplementary Video 4 Neurod6:CherryNTR2.0 predatory behavior at 50-day post DMSO treatment.mp4 Cite Share Download PDF Status: Published Journal Publication published 20 Jan, 2026 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. 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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-6585454","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":452854899,"identity":"dca42c6d-bf16-45a1-a6d3-10551dea2d5b","order_by":0,"name":"Ji-Feng 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Here we establish a Nitroreductase (NTR)-based inducible cell ablation system in axolotls. Through generation of Sox2:Cherry-NTR knock-in axolotls, we achieve efficient ablation of ependymoglial cells (EGCs) in the central nervous system. Combined spinal cord and brain transplantation and injury models demonstrate regeneration failure upon EGC depletion, suggesting that EGCs are solo source of central nervous system regeneration. Additionally, EGC ablation in the spinal cord resulted in delayed tail regeneration. Moreover, we establish NeuroD6:Cherry-NTR and NeuroD6:Cherry-NTR2.0 knock-in lines to ablate postmitotic cortical neurons, to model degenerative disease and study the process of brain regeneration after large scale neuronal depletion. We found that NTR2.0 (but not NTR) leads to elimination of \u003e95% of cortical neurons. Following the depletion of nearly all cortical neurons, all lost neuronal subtypes are chronological regenerated with layer organization mirroring developmental patterning. Finally, we create Cre-LoxP-based conditional NTR2.0 transgenic axolotls using a constitutive CAGGs promoter, enabling tissue-specific ablation of the targeted cell when combine established Cre lines. 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