Tissue-to-Analysis Framework Enables Multiscale Mapping of the Architectural and Cellular Organization in the Human Dorsal Root Ganglion

Abstract

Transcriptomic studies have helped us understand the dorsal root ganglias cellular milieu, yet our knowledge of protein expression and spatial organization/architecture remains less defined. Here we establish a comprehensive resource from processing through analysis of hDRG tissue. We optimize tissue-handling strategies and evaluate 114 antibodies targeting neuronal and non-neuronal cell types, identifying protocols that preserve neuronal morphology and antigen retain specificity. Integrating these workflows with our Deep Learning-assisted image analysis pipelines, we quantify size, expression, and spatial organization across 35,721 neurons from 15 donors. Female donors exhibited significantly larger neuronal somata, indicating sexual dimorphism. Neuronal subpopulations display clear spatial clustering. We further characterized the perineuronal niche, marked by dense vascularization, nuclear remodeling in perineuronal cells, and age-related increased turnover of neuron-associated macrophages. Together, this resource provides standardized methodologies and quantitative frameworks for reproducible protein-level interrogation of human sensory biology and pain mechanisms.