Live imaging of excitable axonal microdomains in ankyrin-G-GFP mice

• DOI: 10.7554/elife.87078.1
• Publication Date: 2023-06-27T10:54:42Z
• Authors: Christian Thome; Jan Maximilian Ja...; Seda Karabulut; Claudio Acuna; Elisa D’Este; Stella J. Soyka; Konrad Baum; Michael Bock; Nadja Lehmann; Masashi Hasegawa; Dan Alin Ganea; Chloé Maëlle Benoit; Jan Gründemann; Christian Schultz; Vann Bennett; Paul M. Jenkins; Maren Engelhardt
• Keywords: Ankyrin; Live cell imaging; Ankyrin repeat

The axon initial segment (AIS) constitutes not only the site of action potential initiation, but also a hub for activity-dependent modulation 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 reporter line which is intrinsically labeled by an ankyrin-G-GFP fusion protein activated Cre recombinase, tagging native Ank3 gene. Using confocal, superresolution, and two-photon microscopy as well whole-cell patch-clamp recordings vitro, ex , confirm that subcellular scaffold electrophysiological parameters labelled cells remain unchanged. We further uncover rapid remodeling following increased network activity this model system, highly reproducible labeling over weeks. This novel allows longitudinal plasticity real time thus provides unique approach to study broad range applications.