Single-cell and isoform-specific translational profiling of the mouse brain

Area of Science:

• Neuroscience
• Molecular Biology
• Genomics

Background:

• The brain utilizes complex post-transcriptional regulation for mRNA translation.
• Alternative splicing is vital for neuronal specificity and implicated in neurological disorders.
• Previous methods lacked single-cell, isoform-sensitive resolution for translatome analysis.

Purpose of the Study:

• To develop and apply an isoform-sensitive, single-cell translatomics platform for the brain.
• To comprehensively map cell-type-specific mRNA translation in the mouse hippocampus.
• To investigate differential translation of gene isoforms across distinct neuronal populations.

Main Methods:

• Deployment of Surveying Ribosomal Targets by APOBEC-Mediated Profiling (Ribo-STAMP).
• Integration with short-read and long-read single-cell RNA sequencing.
• Analysis of mouse hippocampus translatomes at single-cell resolution.

Main Results:

• Generated the first isoform-sensitive single-cell translatomes of the mouse hippocampus.
• Discovered cell-type-specific translation of 3,857 alternative transcripts across 1,641 genes.
• Identified differential translation of isoforms within and across 8 distinct cell types, defining translational states in CA1 and CA3 neurons.

Conclusions:

• The Ribo-STAMP platform enables high-resolution mapping of cell-type and isoform-specific translation in the brain.
• Findings reveal novel insights into the regulation of neuronal function and disease pathogenesis.
• This technology will advance the understanding of translational control in brain physiology and neurological disorders.