Cell-type profiling of the sympathetic nervous system using spatial transcriptomics and spatial mapping of mRNA

Area of Science:

• Neuroscience
• Developmental Biology
• Genomics

Background:

• Identifying neural progenitor cells for sympathetic nervous system (SNS) development is challenging due to limitations in mapping molecular data to tissue architecture.
• Current methods lack the resolution to precisely locate these cells within complex developing neural tissues.

Purpose of the Study:

• To develop and apply a novel spatial transcriptomics strategy for molecularly identifying neural progenitor cells involved in sympathetic nervous system formation.
• To overcome technical limitations in spatial mapping of molecular information within developing neural tissues.

Main Methods:

• Applied Slide-seq spatial transcriptomics to embryonic chick trunk tissue.
• Integrated data with single-cell (sc) RNA-seq and 10× Genomics Visium.
• Utilized HiPlex RNAscope fluorescence in situ hybridization and multispectral confocal microscopy for gene target visualization.

Main Results:

• Successfully identified molecular signatures of neural progenitor cells in peripheral sympathetic ganglia (SG) and spinal cord preganglionic neurons (PGNs).
• Visualized 12 gene targets in precise locations within developing chick trunk tissue.
• Achieved high-resolution mapping of molecular heterogeneity in neural tissue.

Conclusions:

• Demonstrated a robust strategy for integrating single-cell and spatial transcriptomic data.
• Provided a roadmap for functional studies of neural connectivity in the developing sympathetic nervous system.
• Established a platform for addressing complex questions in neural development and regeneration.