Mapping the axolotl brain and its regeneration

The Axolotl is an extraordinary model system to study regeneration in tetrapods. Remarkably, the axolotl isable to regenerate parts of the brain after major injury, however the molecular mechanisms that orchestrateregeneration are unclear, and the completeness of functional circuit and behavioral recovery is unknown. Ourvision is to develop the axolotl model system to rigorously study central nervous system regeneration, anduse discoveries from the axolotl to design novel strategies for regenerating mammalian tissues. The mappingof brain cell types and connections in the axolotl will also have a profound impact on the understanding ofvertebrate brain organization and evolution.In AxoBrain, we will integrate scale-crossing technologies to map the axolotl brain at molecular, cellularand circuit resolution, and establish assays to understand how brain cells respond to damage and behavioralcircuits regenerate. Our project will be achieved through the following objectives: 1) We will generate aspatiotemporal multiomic atlas of the axolotl brain at single cell resolution. 2) We will generate aconnectome of the axolotl retino-tectal-hindbrain circuit and develop behavioral and circuit functional assaysin response to visual stimulation. 3) We will establish injury assays and assess regeneration throughbehavioral, connectomic and single-cell multiomic analysis. 4) Finally, we will apply CRISPR perturbationscreens with single-cell genomic readout in vivo and in vitro to identify modulators of regeneration.AxoBrain utilizes quantitative and state-of-the-art methods to explore how the axolotl can regenerate neuralcircuitry after injury. We have assembled a team with synergistic skill sets that will be required to achieveour vision. This project is a groundbreaking starting point to rigorously investigate how mammals have lostregeneration capabilities over evolution, and will serve as a springboard to design novel strategies forregenerating