NRG1/ErbB signalling controls the dialogue between macrophages and neural crest-derived cells during zebrafish fin regeneration

Béryl Laplace-Builhé,Mai Nguyen-Chi,Gautier Tejedor,Marc Mathieu,Candice Bohaud,Said Assou,Patricia Luz-Crawford,Christian Jorgensen,Audrey Barthelaix,Dany Severac,Farida Djouad,Marine Pratlong

doi:10.1038/s41467-021-26422-5

Abstract

Fish species, such as zebrafish (Danio rerio), can regenerate their appendages after amputation through the formation of a heterogeneous cellular structure named blastema. Here, by combining live imaging of triple transgenic zebrafish embryos and single-cell RNA sequencing we established a detailed cell atlas of the regenerating caudal fin in zebrafish larvae. We confirmed the presence of macrophage subsets that govern zebrafish fin regeneration, and identified a foxd3-positive cell population within the regenerating fin. Genetic depletion of these foxd3-positive neural crest-derived cells (NCdC) showed that they are involved in blastema formation and caudal fin regeneration. Finally, chemical inhibition and transcriptomic analysis demonstrated that these foxd3-positive cells regulate macrophage recruitment and polarization through the NRG1/ErbB pathway. Here, we show the diversity of the cells required for blastema formation, identify a discrete foxd3-positive NCdC population, and reveal the critical function of the NRG1/ErbB pathway in controlling the dialogue between macrophages and NCdC.

Highlights

Fish species, such as zebrafish (Danio rerio), can regenerate their appendages after amputation through the formation of a heterogeneous cellular structure named blastema
To identify the paracrine mechanism underlying foxd3+neural crest-derived cells (NCdC) role in the macrophage response during regeneration, we focused our attention on neuregulin 1 (NRG1), a critical factor for the development of neural crest (NC) cells and of some NCdC, including SC32 that promotes the proliferation of damaged tissue cells in various models and during regeneration[33,34,35]
In Tg(foxd3:mCherry)ct[110] mutants, erbb[2], and erbb[3] expression levels were not different compared with those in WT larvae and in response to amputation (Fig. 5g and Supplementary Fig. 5a). These results suggest that the significant erbb[2] and nrg1.004 upregulation observed in wild-type larvae upon amputation depends on foxd[3], and that foxd3-dependent NRG1/ErbB2 signalling has an essential role in appendage regeneration, by highlighting the correlation between nrg[1] expression and presence of foxd3+ cells in the regenerating blastema

Summary

Introduction

Fish species, such as zebrafish (Danio rerio), can regenerate their appendages after amputation through the formation of a heterogeneous cellular structure named blastema. The list of cell types and factors involved in blastema formation and appendage regeneration has been expanded over the last years, but comparatively little is known about the cells and genes engaged upon tissue amputation in vertebrates To overcome this lack of knowledge, single-cell RNA sequencing (scRNA-seq) and lineage tracing experiments have been recently combined to study the axolotl limb blastema composition[8,9]. These studies allowed clearly demonstrating that the heterogeneous fibroblast population of the blastema loses its adult features, adopts a multipotent skeletal progenitor phenotype, and expresses genes of the developmental-like state[8,9] They confirmed the presence of muscle satellite cells, fibroblasts, and macrophages in the regenerating axolotl limbs, and identified different cell types[9]. These studies suggest that SC might play a pivotal role in the tight regulation of the immune response required for regeneration

Methods

Results

Conclusion