POU6F1 promote lumbar motor circuit reorganization following spinal cord injury

Spinal cord injury (SCI) causes irreversible motor deficits due to disrupted lumbar circuitry. However, transcriptional mechanisms in distal lumbar circuits are poorly understood. We identify POU6F1 as a critical transcriptional regulator in spinal lumbar segment (SLS, L3-L5) motor circuit regeneration. Using an integrative approach combining a T9 contusion SCI model, primary motor neurons (MNs) cultures exposed to OGD/R, RNA sequencing, anterograde/retrograde neural tracing, IncuCyte® ZOOM live-cell imaging, Co-IP, molecular docking, luciferase reporter assays, transmission electron microscopy, and other techniques, we identified sustained POU6F1 upregulation temporally coupled to endogenous SLS MNs axon regeneration after SCI. AAV-mediated POU6F1 overexpression activated JNK/c-Jun/GAP-43/BDNF signaling, rescuing MNs viability and promoting neurite outgrowth, however, these restorative effects were fully abolished by JNK inhibition. POU6F1 overexpression in SLS MNs enhanced dendritic complexity, synaptic reorganization, and the descending propriospinal tract (dPST) axonal growth. This overexpression further attenuated SCI-induced dendritic pathology, drove remodeling of the SLS motor circuit, and culminated in tibialis anterior (TA) muscle reinnervation and locomotor recovery. Mechanistically, POU6F1 directly bound JNK and transactivated the c-Jun promoter, thereby driving downstream effectors GAP-43 and BDNF/TrkB expression to synergistically enhancing intrinsic neurite outgrowth and microenvironmental remodeling. Our findings establish POU6F1 as the first transcription factor coordinating SLS motor circuit reorganization.