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. 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.
Authors
- Chloé M Benoit (ORCID: https://orcid.org/0000-0003-2634-2650)
- Nikolas A Stevens (ORCID: https://orcid.org/0000-0002-5592-3326)
- Stella J Soyka (ORCID: https://orcid.org/0000-0002-8070-8951)
- Johannes Roos (ORCID: https://orcid.org/0000-0001-7409-9101)
- Jan Gründemann (ORCID: https://orcid.org/0000-0002-3149-0001)
- Seda Karabulut (ORCID: https://orcid.org/0000-0003-3302-5004)
- Paul M. Jenkins (ORCID: https://orcid.org/0000-0002-4207-5823)
- Maren Engelhardt (ORCID: https://orcid.org/0000-0001-8020-6604)
- Michael Bock (ORCID: https://orcid.org/0000-0001-9720-3506)
- Dan A Ganea (ORCID: https://orcid.org/0000-0003-2558-8202)
- Kalynn M Bird (ORCID: https://orcid.org/0009-0008-7818-6243)
- Elisa D’Este (ORCID: https://orcid.org/0000-0003-0781-187X)
- Vann Bennett (ORCID: https://orcid.org/0000-0003-2695-7209)
- Konrad Baum
- Nadja Lehmann (ORCID: https://orcid.org/0000-0003-4801-3057)
- Claudio Acuna (ORCID: https://orcid.org/0000-0002-8849-6260)
- Masashi Hasegawa (ORCID: https://orcid.org/0000-0003-4055-4377)
- Christian Thome (ORCID: https://orcid.org/0000-0002-3344-8683)
- Jan Maximilian Janssen (ORCID: https://orcid.org/0000-0002-3818-7910)
- Lia Min (ORCID: https://orcid.org/0000-0002-3977-1684)
- Christian Schultz (ORCID: https://orcid.org/0000-0002-9040-9891)
Institutions
- Johannes Kepler University of Linz (AT)
- University of Basel (CH)
- Heidelberg University (DE)
- University of Michigan (US)
- German Center for Neurodegenerative Diseases (DE)
- University Hospital Heidelberg (DE)
- Max Planck Institute for Medical Research (DE)
- Chica and Heinz Schaller Foundation (DE)
- Duke Medical Center (US)
- Duke University Hospital (US)
- Czech Academy of Sciences, Institute of Physiology (CZ)
Publication Details
- Journal
- eLife
- Published
- 2025-02-03
- DOI
- https://doi.org/10.7554/elife.87078.3
- Citations
- 4
- Primary Topic
- Erythrocyte Function and Pathophysiology
- Type
- article
- Field-Weighted Citation Impact
- 4.66
Funders
- Deutsche Forschungsgemeinschaft
- Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung
- National Institutes of Health