Transcriptional profiling of mouse peripheral nerves to the single-cell level to build a sciatic nerve ATlas (SNAT).

Daniel Gerber,Emilio Yángüez,Jorge A Pereira,Joanne Gerber,Ge Tan,Slavica Dimitrieva,Ueli Suter

doi:10.7554/elife.58591

Abstract

Peripheral nerves are organ-like structures containing diverse cell types to optimize function. This interactive assembly includes mostly axon-associated Schwann cells, but also endothelial cells of supporting blood vessels, immune system-associated cells, barrier-forming cells of the perineurium surrounding and protecting nerve fascicles, and connective tissue-resident cells within the intra-fascicular endoneurium and inter-fascicular epineurium. We have established transcriptional profiles of mouse sciatic nerve-inhabitant cells to foster the fundamental understanding of peripheral nerves. To achieve this goal, we have combined bulk RNA sequencing of developing sciatic nerves up to the adult with focused bulk and single-cell RNA sequencing of Schwann cells throughout postnatal development, extended by single-cell transcriptome analysis of the full sciatic nerve both perinatally and in the adult. The results were merged in the transcriptome resource Sciatic Nerve ATlas (SNAT: https://www.snat.ethz.ch). We anticipate that insights gained from our multi-layered analysis will serve as valuable interactive reference point to guide future studies.

Highlights

Peripheral nerves are specialized in the transport of information between the central nervous system and peripheral body parts, including muscles and sensory cells (Stassart et al, 2018)
We developed a strategy that allowed the distinction of two fractions of Schwann cells (SCs) from postnatal day 5 (P5) sciatic nerves by Fluorescence-Activated Cell Sorting (FACS), one enriched in myelinating SCs and the another enriched in not-myelinating SCs
We started by resolving the transcriptomes of peripheral nerves at various time points of nerve maturation and SC differentiation using bulk RNA sequencing technology

Summary

Introduction

Peripheral nerves are specialized in the transport of information between the central nervous system and peripheral body parts, including muscles and sensory cells (Stassart et al, 2018). Exemplary potential benefits include: (1) Straight-forward determination of transcript expression and changes during development and in the adult nerve, in bulk and down to the cellular level; (2) Potential use of gene expression in individual cells, or in group of cells, to expand and refine the current set of available markers that can be employed to define cells with particular structures and functions in the nerve tissue under different conditions in health and disease; (3) Identification of regulated genes that may provide the basis for developing improved tools for cell-type specific labeling and genetic alteration studies; and (4) Refinement of the interpretation and integration of previous research results that were obtained with lower spatial and/or temporal cellular resolution, or with different experimental emphasis. Interpretations, and data mining, we merged our datasets in an openly accessible web-based resource called Sciatic Nerve ATlas (SNAT) at https://www.snat.ethz.ch/

Results and discussion

Materials and methods

Data and code availability