TY  - JOUR
AU  - Thome, Christian
AU  - Janssen, Jan Maximilian
AU  - Karabulut, Seda
AU  - Acuna, Claudio
AU  - D'Este, Elisa
AU  - Soyka, Stella J
AU  - Baum, Konrad
AU  - Bock, Michael
AU  - Lehmann, Nadja
AU  - Roos, Johannes
AU  - Stevens, Nikolas A
AU  - Hasegawa, Masashi
AU  - Ganea, Dan Alin
AU  - Benoit, Chloé M
AU  - Gründemann, Jan
AU  - Min, Lia Y
AU  - Bird, Kalynn M
AU  - Schultz, Christian
AU  - Bennett, Vann
AU  - Jenkins, Paul
AU  - Engelhardt, Maren
TI  - Live imaging of excitable axonal microdomains in ankyrin-G-GFP mice
JO  - eLife
VL  - 12
SN  - 2050-084X
CY  - Cambridge
PB  - eLife Sciences Publications
M1  - DZNE-2025-00294
SP  - RP87078
PY  - 2025
AB  - 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.
KW  - Animals
KW  - Ankyrins: metabolism
KW  - Ankyrins: genetics
KW  - Mice
KW  - Green Fluorescent Proteins: metabolism
KW  - Green Fluorescent Proteins: genetics
KW  - Axons: physiology
KW  - Axons: metabolism
KW  - Axon Initial Segment: metabolism
KW  - Patch-Clamp Techniques
KW  - Genes, Reporter
KW  - Mice, Transgenic
KW  - Action Potentials
KW  - AIS plasticity (Other)
KW  - ankryin-G (Other)
KW  - axon initial segment (Other)
KW  - live imaging (Other)
KW  - mouse (Other)
KW  - neuroscience (Other)
KW  - node of Ranvier (Other)
KW  - Ankyrins (NLM Chemicals)
KW  - Green Fluorescent Proteins (NLM Chemicals)
KW  - Ank3 protein, mouse (NLM Chemicals)
LB  - PUB:(DE-HGF)16
C6  - pmid:39898808
DO  - DOI:10.7554/eLife.87078
UR  - https://pub.dzne.de/record/276472
ER  -