Adi powerlab 8 30 stimulator

At 117 days of age, mice were anesthetized with isoflurane (Piramal Healthcare, Bethlehem, Pennsylvania) at a concentration of 1.0–1.5% in oxygen. Animals were placed supine, and the abdomen was shaved and cleaned with 70% ethanol. Phrenic nerve conduction studies were performed with stimulation of the phrenic nerve via needle electrodes trans-cutaneously inserted into the neck region in proximity to the passage of the phrenic nerve (Cheng et al., 2021 (link); Ghosh et al., 2018 (link)). A reference electrode was placed on the shaved surface of the right costal region. Phrenic nerve was stimulated with a single burst at 6 mV (amplitude) for a 0.5 ms duration. Each animal was stimulated between 10 and 20 times to ensure reproducibility, and recordings were averaged for analysis. ADI Powerlab8/30stimulator and BioAMPamplifier (ADInstruments, Colorado Springs, CO) were used for both stimulation and recording, and Scope 3.5.6 software (ADInstruments, Colorado Springs, CO; RRID: SCR_001620) was used for subsequent data analysis. Following recordings, animals were immediately euthanized, and tissue was collected (as described below).

Four weeks post-surgery, rats were anesthetized with isoflurane (Piramal Healthcare, Bethlehem, PA, United States) at a concentration of 3.0–3.5% diluted in oxygen. Animals were placed supine and the region just below the rib cage was shaved and cleaned with 70% ethanol. Phrenic nerve conduction studies were performed with stimulation at the neck via near nerve needle electrodes placed along the phrenic nerve. A reference electrode was placed on the shaved surface of the right costal region. The phrenic nerve was stimulated with a single burst at 6 mV (amplitude) for a 0.5 ms duration. Each animal was stimulated between 10 and 20 times to ensure reproducibility, and recordings were averaged for analysis. Animals were daily followed for any signs of distress in response to this procedure. ADI Powerlab 8/30 stimulator and BioAMPamplifier (ADInstruments, Colorado Springs, CO, United States) were used for both stimulation and recording, and Scope 3.5.6 software (ADInstruments, Colorado Springs, CO, United States) was used for subsequent data analysis. An additional control animal without lesion (laminectomy only) was used as an example of a normal CMAP recording.

Rats were anesthetized with isoflurane (Piramal Healthcare) at a concentration of 3.0–3.5% diluted in oxygen. Animals were placed supine, and the region just below the rib cage was shaved and cleaned with 70% ethanol. Phrenic nerve conduction studies were performed with stimulation of the phrenic nerve via needle electrodes transcutaneously inserted into the neck of the rat in proximity to the passage of the phrenic nerve (Lepore et al., 2008 (link), 2010 (link)). A reference electrode was placed on the shaved surface of the right costal region. The phrenic nerve was stimulated with a single burst at 6 mV (amplitude) for a 0.5-ms duration. Each animal was stimulated between 10 and 20 times to ensure reproducibility, and recordings were averaged for analysis. ADI Powerlab8/30 stimulator and BioAMP amplifier (ADInstruments) were used for both stimulation and recording, and Scope 3.5.6 software (ADInstruments; RRID: SCR_001620) was used for subsequent data analysis. Following recordings, animals were immediately euthanized with a dose of Euthasol, and tissue was collected (as described below).