Reversible cervical hemispinalization of the rat spinal cord by a cooling device

Differentiation between traumatic and activity dependent plasticity in the CNS has been a challenge to neuroscientists in the past. We describe a cooling device that allows reversible block of the inspiratory drive to phrenic motoneurons without injury to the spinal cord at the C2 level. Thus, this experimental approach can be used to differentiate between the plasticity induced by blockade of synaptic activity in the phrenic nucleus from the trauma-induced plasticity caused by a C2 spinal cord hemisection which would also interrupt descending inspiratory drive. Complete block of axon transmission of the respiratory pathways running unilaterally in the ventral as well as in the lateral funiculus was achieved by approximation of a cold probe to the ventral surface of the spinal cord. The spinal cord surface temperature was lowered to 7 degrees C. The temperature was maintained by a cold recirculated alcohol system. The efficacy of the reversible block was assessed by bilateral continuous EMG activity recording from the hemidiaphragms ipsilateral and contralateral to the cold application. Quantitative analysis of the EMG hemidiaphragmatic signals was performed in two sham-operated control (no cold application) and an experimental (cold application) group of Sprague-Dawley rats. The control groups were employed to confirm that the surgical exposure of the cord and/or the chronic placement of the probe and the administration of IV dopamine given to maintain stable blood pressure did not affect respiration. No significant change occurred in EMG hemidiaphragmatic activity in control animals. The descending pathway from the rVRG to the phrenic nucleus was completely and continuously blocked for 4 h in all four experimental animals as demonstrated by abolition of the EMG hemidiaphragmatic signal ipsilateral to cold block. In all experimental animals hemidiaphragmatic activity returned when the cold block was removed. The recovered EMG activity was significantly higher than pre-block values. Interestingly, EMG activity contralateral to the block did not change significantly from control values after the block was removed, but was significantly enhanced during cold block. The present results suggest that cold block provides a means of studying activity-dependent plasticity in the respiratory pathways of the spinal cord.