Massive sequential spatial transcriptional RNA sequencing by capturing mouse spinal cord tissue sections

Abstract Objectives: To prepare for the analysis of spatio-temporal heterogeneity in the pathological process of spinal cord injury, by capturing sections of mouse spinal cord tissue, RNA sequencing was performed for massive sequential spatial transcriptional.Background: With the innovation of sequencing technology, probing the gene expression of an individual cell enables the identification of cell characteristics, type and state. Single-cell RNA sequencing (scRNA-seq) systematically identifies cell populations in tissues, but characterization of their spatial architectures remains challenging. Spatial transcriptomics assays gene expression at the RNA level, while preserving the valuable spatial context of data.Methods: T9-T11 thoracic spinal cord tissue sections were captured from mice suffering spinal cord injury for downstream massive sequential spatial transcriptional RNA sequencing by utilizing Poly-A capture and unique spatial barcodes to achieve unbiased gene expression at high spatial resolution.Results: Different sample libraries in four capture regions were constructed, and a total of 16 spatio-temporal heterogeneous spinal cord injury samples after data separation were successfully captured.Conclusions: The 10x Genomics Visium platform enables to obtain frozen sections from fresh or frozen stored spinal cord samples to study spatial-temporal characteristics of cells clustering, investigates heterogeneity and signaling pathways within spinal cords, which helps to augment ongoing neuroscience and spatial transcriptomics research.