Veterinary multi channel anesthesia delivery system and vaporizer

According to our published protocol (11 (link)), VPA (Calbiochem, San Diego, Calif) solution was prepared fresh by dissolving it in filtered distilled water to create a 400-mg/mL solution. Anesthesia was induced with 5% isoflurane (Abbott Laboratories, North Chicago, Ill) mixed with air in an induction chamber. Rats were then fitted with a nose cone scavenging system and allowed to breathe spontaneously. Anesthesia was maintained by delivering 0.7% to 1.2% isoflurane via the nose cone using a veterinary multichannel anesthesia delivery system and vaporizer (Kent Scientific Corporation, Torrington, Ct). After injecting 0.2 mL of 0.5% bupivacaine (AstraZeneca, Wilmington, Del) for local anesthesia, an incision was made over the right femoral vessels; the femoral artery and vein were dissected, and both vessels were cannulated with polyethylene 50 (PE50) catheters (Clay Adams, Sparks, Md). The venous cannula was used for hemorrhage, whereas the arterial catheter was connected to the Ponemah Physiology Platform (Gould Instrument Systems, Valley View, Ohio) for continuous hemodynamic monitoring in the first hit of HS.

On the day of experimentation, Tubastatin-A (70mg/kg, Calbiochem, San Diego, CA) solution was prepared freshly by dissolving it in dimethyl sulfoxide (DMSO; 1μl/g animal body weight). ^{24 (link)} Anesthesia was induced with 4% isoflurane (Abbott Laboratories, North Chicago, IL) mixed with air in an induction chamber, and maintained by delivering 0.8–1.5% isoflurane via the nose cone using a veterinary multi-channel anesthesia delivery system and vaporizer (Kent Scientific Corporation, Torrington, CT). Body temperature was maintained with an automated heating pad by monitoring anus temperature. After injecting 0.2mL of 0.25% bupivacaine (APP pharmaceuticals, LLC. Schaumburg, IL) for local anesthesia, an incision was made over the left femoral vessels. The femoral artery was dissected and cannulated with polyethylene 50 catheters (Clay Adams, Sparks, MD) for creating hemorrhage, obtaining blood samples, and hemodynamic monitoring (Ponemah Physiology Platform, Gould Instrument Systems, Valley View, OH).

On the day of experimentation, Tub A (70mg/kg, Calbiochem, San Diego, CA) solution was prepared by dissolving it in dimethylsulfoxide (DMSO, 1μl/gram of body weight) (Sigma-Aldrich, St. Louis, MO). Anesthesia was induced with 4% isoflurane (Abbott Laboratories, North Chicago, IL) mixed with air in an induction chamber, and maintained by delivering 0.8–1.5% isoflurane via the nose cone using a veterinary multi-channel anesthesia delivery system and vaporizer (Kent Scientific Corporation, Torrington, CT). Core body temperature was maintained with an automated heating pad. Bupivacaine (0.2mL of 0.25%, APP pharmaceuticals, LLC. Schaumburg, IL) was injected for local anesthesia, and using a micro cutdown technique femoral artery was cannulated with polyethylene 50 catheter (Clay Adams, Sparks, MD), which was used for blood withdrawal and hemodynamic monitoring (Ponemah Physiology Platform, Gould Instrument Systems, Valley View, OH).