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

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 predominantlypost hocapproaches since no robust murinein vivolive 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 nativeAnk3gene. Using confocal, superresolution, and two-photon microscopy as well as whole-cell patch-clamp recordingsin vitro, ex vivo, andin vivo, we confirm that the subcellular scaffold of the AIS and electrophysiological parameters of labelled cells remain unchanged. We further uncover rapid AIS remodeling following increased network activity in this model system, as well as highly reproduciblein vivolabeling of AIS over weeks. This novel reporter line allows longitudinal studies of AIS modulation and plasticityin vivoin real time and thus provides a unique approach to study subcellular plasticity in a broad range of applications.