Singular strategies for promoting axon regeneration and motor recovery after spinal cord injury (SCI) have been attempted with limited success. For instance, the deletion of RhoA and Pten (an extrinsic and intrinsic modulating factor, respectively) in corticospinal neurons promotes axon sprouting after thoracic SCI, however it is unable to restore motor function. Here, we examine the effects of combining RhoA/Pten deletion in corticospinal neurons with chemogenetic neuronal stimulation on axonal growth and motor recovery after SCI in mice. We find that this combinatorial approach promotes greater axonal growth and synaptic bouton formation in corticospinal neurons within the spinal cord compared to RhoA;Pten deletion alone. Furthermore, chemogenetic neuronal stimulation of RhoA;Pten-deleted corticospinal neurons improved forelimb performance in behavioral tasks after SCI compared to RhoA;Pten deletion alone. These results demonstrate that combination therapies pairing genetic modifications with neuronal stimulation can promote greater synaptic formation and motor recovery following SCI than either strategy alone.Significance Statement In this study, we examined whether pairing neural stimulation with genetic deletion of RhoA and Pten in corticospinal neurons would enhance axonal growth and motor recovery after spinal cord injury (SCI). We found that this combinatorial approach, relative to singular strategies, promoted greater presynaptic bouton formation in injured corticospinal neurons in the spinal cord, resulting in improved motor recovery in those mice. These results open the possibility for even greater enhancements in healing and recovery following SCI through carefully crafted, multifaceted treatments tailored to address the specific needs of individual SCI patients.
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