Dissecting the Mechanisms of Neuromuscular Junction Plasticity through a Single-Cell Multi-Omics Lens

Abstract

The neuromuscular junction (NMJ) serves as the fundamental interface between motor neurons and skeletal muscles, enabling precise motor control. Its structural and functional plasticity underlies muscle regeneration, motor learning, and recovery from injury. Traditional approaches relying on histology and bulk omics have provided valuable insights into NMJ structure but fail to capture cell-type heterogeneity, temporal dynamics, and spatial coordination during regeneration. A systems-level understanding of NMJ plasticity remains limited. This study integrates single-cell RNA sequencing, proteomic, and spatial transcriptomic data to construct a multidimensional framework for NMJ plasticity. Analytical methods include clustering, trajectory inference, ligand-receptor network modeling, and cross-omic module identification, interpreted under the Neuro-Muscular Plasticity Systems Framework (NMPSF). The analysis reveals specialized Schwann cell subtypes, bidirectional neuron-muscle communication through neuregulin-ErbB and agrin-MuSK pathways, and spatially organized extracellular matrix remodeling supporting reinnervation. Integrated multi-omic networks further identify molecular modules governing synaptic stability, regeneration, and immune-ECM interaction. These findings redefine NMJ plasticity as a coordinated, multi-level process linking molecular signaling, cellular diversity, and spatial organization. The proposed NMPSF offers a reproducible analytical and conceptual framework for studying neuromuscular adaptation and guiding regenerative interventions for neuromuscular disorders.