Neuropilin-1-mediated pruning of corticospinal tract fibers is required for motor recovery after spinal cord injury

Abstract

Following incomplete spinal cord injury (SCI), reorganization of the corticospinal tract (CST) contributes to spontaneous motor recovery. Axotomized CST fibers form collaterals and make synapses with interneurons, followed by pruning of excess fibers. Although axonal pruning is involved in refinement of neural circuits, its molecular mechanisms and functional roles remain poorly understood. To address these questions, we performed dorsal hemisections of mouse thoracic spinal cord. We observed that Neuropilin-1 (Nrp1) mRNA was upregulated in layer 5 pyramidal neurons in the motor cortex 14 days after SCI, when the pruning occurred. Nrp1 knockdown using adeno-associated virus (AAV) vector encoding Nrp1 shRNA in the hindlimb motor area impaired the pruning of collaterals after SCI. Nrp1 knockout by injecting AAV vector encoding Cre recombinase into Nrp1 floxed mice also suppressed axonal pruning. Propriospinal neurons, interneurons that connect CST and motoneurons, expressed Semaphorin 3A (Sema3A), the ligand of Nrp1. Furthermore, the genetic deletion of Nrp1 specifically in the hindlimb motor area suppressed the recovery of skilled movement at 21 and 28 days after SCI. The present findings demonstrate that the pruning of collaterals mediated by Nrp1 is required for motor recovery after SCI, and suggest that refinement of the neuronal network facilitates motor recovery.