Live imaging of excitable axonal microdomains in ankyrin-G-GFP mice
The axon initial segment (AIS) constitutes not only the site of action potential initiation, but also a hub for activity-dependent modulation of output generation. Recent studies shedding light on AIS function used predominantly post-hoc approaches since no robust murine in vivo live reporters exist...
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eLife Sciences Publications Ltd
2025-02-01
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| Online Access: | https://elifesciences.org/articles/87078 |
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| author | Christian Thome Jan Maximilian Janssen Seda Karabulut Claudio Acuna Elisa D'Este Stella J Soyka Konrad Baum Michael Bock Nadja Lehmann Johannes Roos Nikolas A Stevens Masashi Hasegawa Dan A Ganea Chloé M Benoit Jan Gründemann Lia Y Min Kalynn M Bird Christian Schultz Vann Bennett Paul M Jenkins Maren Engelhardt |
| author_facet | Christian Thome Jan Maximilian Janssen Seda Karabulut Claudio Acuna Elisa D'Este Stella J Soyka Konrad Baum Michael Bock Nadja Lehmann Johannes Roos Nikolas A Stevens Masashi Hasegawa Dan A Ganea Chloé M Benoit Jan Gründemann Lia Y Min Kalynn M Bird Christian Schultz Vann Bennett Paul M Jenkins Maren Engelhardt |
| author_sort | Christian Thome |
| collection | DOAJ |
| description | The axon initial segment (AIS) constitutes not only the site of action potential initiation, but also a hub for activity-dependent modulation of output generation. Recent studies shedding light on AIS function used predominantly post-hoc approaches since no robust murine in vivo live reporters exist. Here, we introduce a reporter line in which the AIS is intrinsically labeled by an ankyrin-G-GFP fusion protein activated by Cre recombinase, tagging the native Ank3 gene. Using confocal, superresolution, and two-photon microscopy as well as whole-cell patch-clamp recordings in vitro, ex vivo, and in vivo, we confirm that the subcellular scaffold of the AIS and electrophysiological parameters of labeled cells remain unchanged. We further uncover rapid AIS remodeling following increased network activity in this model system, as well as highly reproducible in vivo labeling of AIS over weeks. This novel reporter line allows longitudinal studies of AIS modulation and plasticity in vivo in real-time and thus provides a unique approach to study subcellular plasticity in a broad range of applications. |
| format | Article |
| id | doaj-art-ed7e6a316fab4127895becdece2bebfe |
| institution | Kabale University |
| issn | 2050-084X |
| language | English |
| publishDate | 2025-02-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj-art-ed7e6a316fab4127895becdece2bebfe2025-02-05T17:07:00ZengeLife Sciences Publications LtdeLife2050-084X2025-02-011210.7554/eLife.87078Live imaging of excitable axonal microdomains in ankyrin-G-GFP miceChristian Thome0https://orcid.org/0000-0002-3344-8683Jan Maximilian Janssen1Seda Karabulut2Claudio Acuna3Elisa D'Este4Stella J Soyka5Konrad Baum6Michael Bock7Nadja Lehmann8https://orcid.org/0000-0003-4801-3057Johannes Roos9Nikolas A Stevens10https://orcid.org/0000-0002-5592-3326Masashi Hasegawa11Dan A Ganea12Chloé M Benoit13Jan Gründemann14Lia Y Min15https://orcid.org/0000-0002-3977-1684Kalynn M Bird16Christian Schultz17Vann Bennett18Paul M Jenkins19https://orcid.org/0000-0002-4207-5823Maren Engelhardt20https://orcid.org/0000-0001-8020-6604Institute of Anatomy and Cell Biology, Johannes Kepler University, Linz, Austria; Clinical Research Institute for Neurosciences, Johannes Kepler University, Linz, Austria; Institute of Physiology and Pathophysiology, Heidelberg University, Heidelberg, GermanyInstitute of Anatomy and Cell Biology, Johannes Kepler University, Linz, Austria; Clinical Research Institute for Neurosciences, Johannes Kepler University, Linz, Austria; Institute of Neuroanatomy, Mannheim Center for Translational Neuroscience (MCTN), Medical Faculty Mannheim, Heidelberg University, Mannheim, GermanyInstitute of Neuroanatomy, Mannheim Center for Translational Neuroscience (MCTN), Medical Faculty Mannheim, Heidelberg University, Mannheim, GermanyChica and Heinz Schaller Research Group, Institute of Anatomy and Cell Biology, Heidelberg University, Heidelberg, GermanyOptical Microscopy Facility, Max Planck Institute for Medical Research, Heidelberg, GermanyInstitute of Anatomy and Cell Biology, Dept. of Functional Neuroanatomy, Heidelberg University, Heidelberg, GermanyInstitute of Anatomy and Cell Biology, Johannes Kepler University, Linz, Austria; Clinical Research Institute for Neurosciences, Johannes Kepler University, Linz, AustriaInstitute of Neuroanatomy, Mannheim Center for Translational Neuroscience (MCTN), Medical Faculty Mannheim, Heidelberg University, Mannheim, GermanyInstitute of Neuroanatomy, Mannheim Center for Translational Neuroscience (MCTN), Medical Faculty Mannheim, Heidelberg University, Mannheim, GermanyInstitute of Anatomy and Cell Biology, Johannes Kepler University, Linz, Austria; Clinical Research Institute for Neurosciences, Johannes Kepler University, Linz, Austria; Institute of Neuroanatomy, Mannheim Center for Translational Neuroscience (MCTN), Medical Faculty Mannheim, Heidelberg University, Mannheim, GermanyInstitute of Physiology and Pathophysiology, Heidelberg University, Heidelberg, GermanyGerman Center for Neurodegenerative Disease (DZNE), Neural Circuit Computations, Bonn, GermanyDepartment of Biomedicine, University of Basel, Basel, SwitzerlandGerman Center for Neurodegenerative Disease (DZNE), Neural Circuit Computations, Bonn, Germany; Department of Biomedicine, University of Basel, Basel, SwitzerlandGerman Center for Neurodegenerative Disease (DZNE), Neural Circuit Computations, Bonn, Germany; Department of Biomedicine, University of Basel, Basel, SwitzerlandDepartment of Pharmacology, University of Michigan Medical School, Ann Arbor, United StatesDepartment of Pharmacology, University of Michigan Medical School, Ann Arbor, United StatesInstitute of Neuroanatomy, Mannheim Center for Translational Neuroscience (MCTN), Medical Faculty Mannheim, Heidelberg University, Mannheim, GermanyDepartment of Biochemistry, Duke University Medical Center, Durham, United StatesDepartment of Pharmacology, University of Michigan Medical School, Ann Arbor, United States; Department of Psychiatry, University of Michigan Medical School, Ann Arbor, United StatesInstitute of Anatomy and Cell Biology, Johannes Kepler University, Linz, Austria; Clinical Research Institute for Neurosciences, Johannes Kepler University, Linz, Austria; Institute of Neuroanatomy, Mannheim Center for Translational Neuroscience (MCTN), Medical Faculty Mannheim, Heidelberg University, Mannheim, GermanyThe axon initial segment (AIS) constitutes not only the site of action potential initiation, but also a hub for activity-dependent modulation of output generation. Recent studies shedding light on AIS function used predominantly post-hoc approaches since no robust murine in vivo live reporters exist. Here, we introduce a reporter line in which the AIS is intrinsically labeled by an ankyrin-G-GFP fusion protein activated by Cre recombinase, tagging the native Ank3 gene. Using confocal, superresolution, and two-photon microscopy as well as whole-cell patch-clamp recordings in vitro, ex vivo, and in vivo, we confirm that the subcellular scaffold of the AIS and electrophysiological parameters of labeled cells remain unchanged. We further uncover rapid AIS remodeling following increased network activity in this model system, as well as highly reproducible in vivo labeling of AIS over weeks. This novel reporter line allows longitudinal studies of AIS modulation and plasticity in vivo in real-time and thus provides a unique approach to study subcellular plasticity in a broad range of applications.https://elifesciences.org/articles/87078ankryin-Gaxon initial segmentAIS plasticitylive imagingnode of Ranvier |
| spellingShingle | Christian Thome Jan Maximilian Janssen Seda Karabulut Claudio Acuna Elisa D'Este Stella J Soyka Konrad Baum Michael Bock Nadja Lehmann Johannes Roos Nikolas A Stevens Masashi Hasegawa Dan A Ganea Chloé M Benoit Jan Gründemann Lia Y Min Kalynn M Bird Christian Schultz Vann Bennett Paul M Jenkins Maren Engelhardt Live imaging of excitable axonal microdomains in ankyrin-G-GFP mice eLife ankryin-G axon initial segment AIS plasticity live imaging node of Ranvier |
| title | Live imaging of excitable axonal microdomains in ankyrin-G-GFP mice |
| title_full | Live imaging of excitable axonal microdomains in ankyrin-G-GFP mice |
| title_fullStr | Live imaging of excitable axonal microdomains in ankyrin-G-GFP mice |
| title_full_unstemmed | Live imaging of excitable axonal microdomains in ankyrin-G-GFP mice |
| title_short | Live imaging of excitable axonal microdomains in ankyrin-G-GFP mice |
| title_sort | live imaging of excitable axonal microdomains in ankyrin g gfp mice |
| topic | ankryin-G axon initial segment AIS plasticity live imaging node of Ranvier |
| url | https://elifesciences.org/articles/87078 |
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