eLife (Feb 2025)
Live imaging of excitable axonal microdomains in ankyrin-G-GFP mice
- 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
Affiliations
- Christian Thome
- ORCiD
- Institute 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, Germany
- Jan Maximilian Janssen
- Institute 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, Germany
- Seda Karabulut
- Institute of Neuroanatomy, Mannheim Center for Translational Neuroscience (MCTN), Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany
- Claudio Acuna
- Chica and Heinz Schaller Research Group, Institute of Anatomy and Cell Biology, Heidelberg University, Heidelberg, Germany
- Elisa D'Este
- Optical Microscopy Facility, Max Planck Institute for Medical Research, Heidelberg, Germany
- Stella J Soyka
- Institute of Anatomy and Cell Biology, Dept. of Functional Neuroanatomy, Heidelberg University, Heidelberg, Germany
- Konrad Baum
- Institute of Anatomy and Cell Biology, Johannes Kepler University, Linz, Austria; Clinical Research Institute for Neurosciences, Johannes Kepler University, Linz, Austria
- Michael Bock
- Institute of Neuroanatomy, Mannheim Center for Translational Neuroscience (MCTN), Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany
- Nadja Lehmann
- ORCiD
- Institute of Neuroanatomy, Mannheim Center for Translational Neuroscience (MCTN), Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany
- Johannes Roos
- Institute 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, Germany
- Nikolas A Stevens
- ORCiD
- Institute of Physiology and Pathophysiology, Heidelberg University, Heidelberg, Germany
- Masashi Hasegawa
- German Center for Neurodegenerative Disease (DZNE), Neural Circuit Computations, Bonn, Germany
- Dan A Ganea
- Department of Biomedicine, University of Basel, Basel, Switzerland
- Chloé M Benoit
- German Center for Neurodegenerative Disease (DZNE), Neural Circuit Computations, Bonn, Germany; Department of Biomedicine, University of Basel, Basel, Switzerland
- Jan Gründemann
- German Center for Neurodegenerative Disease (DZNE), Neural Circuit Computations, Bonn, Germany; Department of Biomedicine, University of Basel, Basel, Switzerland
- Lia Y Min
- ORCiD
- Department of Pharmacology, University of Michigan Medical School, Ann Arbor, United States
- Kalynn M Bird
- Department of Pharmacology, University of Michigan Medical School, Ann Arbor, United States
- Christian Schultz
- Institute of Neuroanatomy, Mannheim Center for Translational Neuroscience (MCTN), Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany
- Vann Bennett
- Department of Biochemistry, Duke University Medical Center, Durham, United States
- Paul M Jenkins
- ORCiD
- Department of Pharmacology, University of Michigan Medical School, Ann Arbor, United States; Department of Psychiatry, University of Michigan Medical School, Ann Arbor, United States
- Maren Engelhardt
- ORCiD
- Institute 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, Germany
- DOI
- https://doi.org/10.7554/eLife.87078
- Journal volume & issue
-
Vol. 12
Abstract
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.
Keywords
