Implanted and naive mice were anesthetized and placed on a warmed surgical platform in the supine position and instrumented with ECG leads. The right external jugular vein was isolated by blunt dissection and two sutures were placed rostrally and caudally. The rostral suture was ligated to prevent bleeding. After occluding blood flow by pulling on the caudal suture, a small incision was made in the jugular vein and a 1 French catheter (Instech Labs, Plymouth Meeting, PA) was carefully advanced into the vessel after removing the caudal suture. A small amount of saline was injected to confirm the catheter was functional. The right CS was then exposed. A 1.4 French pressure catheter (SPR-671, Millar, Houston, TX) was carefully advanced into the artery using the same technique described for the jugular vein. Once the two catheters were in place and confirmed functional, 100 μl of phenylephrine (25 μg/kg) supplemented with heparin (7 U/ml) in saline was injected into the jugular vein over 7 s (Fleming et al., 2013) and pressure and HR monitored. Systolic and diastolic pressure and ECG signals were amplified using Bio-Amp Octal (ADInstruments) as described before. To calculate the baroreflex sensitivity index, a 10 s window around the peak systolic blood pressure (BP) was identified to calculate the systolic BP and corresponding HR after phenylephrine injection. Baseline values were calculated from a 10 s window immediately before the injection.

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