Chemogenetic activation of the diaphragm after spinal cord injury in rats

We tested the hypothesis that activation of DREADDs in the mid-cervical spinal cord could restore diaphragm activation during spontaneous breathing after cervical spinal cord injury (SCI). Adult Sprague Dawley rats (n = 7) received bilateral mid-cervical ventral horn injections of an AAV construct encoding an excitatory DREADD (AAV9-hSyn-HA-hM3D(Gq)-mCherry; titer: 2.44 × 10¹³ vg/mL). Subsequently, diaphragm electromyogram (EMG) activity was recorded during spontaneous breathing under isoflurane anesthesia. The selective DREADD ligand JHU37160 (J60) was administered intravenously at acute (3 days), sub-acute (2 weeks), and chronic (2 months) timepoints following cervical hemilesion at spinal level C2. J60 administration resulted in robust increases in diaphragm EMG output at all timepoints, and near-complete restoration of diaphragm EMG activity from the paralyzed hemi-diaphragm in 50 % of trials. Administration of J60 to DREADD naïve, spinal intact rats (n = 8) did not produce an increase in diaphragm activity. These proof-of-concept results indicate that refinement of this technique may provide a strategy for improving diaphragm activation after cervical SCI.