Anesthesia was induced with 3% isoflurane gas followed by ketamine (100mg/kg, IP) and xylazine (5mg/kg, IM). Additional doses of ketamine (20mg/kg IM) were used as needed. Six 0.7-mm diameter holes were drilled over the CFA contralateral to the dominant forelimb at anteroposterior +1.5, +0.5, and −0.5 mm and mediolateral +2.5 and +3.5 mm from bregma.14 (link) To induce cortical ischemia, 0.33 µL of endothelin-1 (ET-1; Bachem Laboratories, 0.3mg/mL) was injected into each hole at a depth of 1.5mm from the cortical surface, through a micropipette (160µm o.d.) attached to a Hamilton syringe using a microsyringe injector (UltraMicro Pump III, World Precision Instruments). Appropriate postoperative care was provided under veterinary supervision.
Protocol detail
Cortical Ischemia Induction Protocol
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Anesthesia
Cortical
Endothelin 1
Forelimb
Ischemia
Isoflurane
Ketamine
Postoperative care
Supervision
Syringe
Xylazine
Corresponding Organization :
Other organizations : Osaka University, University of Kansas Medical Center
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Variable analysis
independent variables
- Anesthesia induced with 3% isoflurane gas
- Ketamine (100mg/kg, IP)
- Xylazine (5mg/kg, IM)
- Additional doses of ketamine (20mg/kg IM) as needed
- Injection of 0.33 µL of endothelin-1 (ET-1; 0.3mg/mL) into six 0.7-mm diameter holes over the CFA contralateral to the dominant forelimb
- Cortical ischemia
- Six 0.7-mm diameter holes drilled over the CFA contralateral to the dominant forelimb at anteroposterior +1.5, +0.5, and −0.5 mm and mediolateral +2.5 and +3.5 mm from bregma
- Injection depth of 1.5mm from the cortical surface
- Appropriate postoperative care under veterinary supervision
- Negative control: No information provided about a negative control group.
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