Stereotaxic Injection of AAVrg-CAG-GFP in Rat Cortex

Rats were handled for two days prior to surgery. Rats were anaesthetized with ketamine (87 mg/kg) and xylazine (10 mg/kg), and they were placed in a stereotaxic apparatus (Stoelting, Woodale, IL) and received a unilateral injection of AAVrg-CAG-GFP (Addgene,^{30 (link)}) into the IL (left- and right-side injections were counterbalanced). In order to perform the injection, a 22-gauge cannula was lowered into place (AP: + 3.00, ML: ± 0.6, DV: − 5.2). A 28-gauge internal cannula (connected to an infusion pump via PE 20 tubing) was inserted into the guide cannula and used to deliver 0.6 μL of the virus at a rate of 0.15 μL per minute and stayed in place for five minutes after the infusion was complete. After suturing rats were injected with Meloxicam (1 mg/kg) and they were returned to their home cages once ambulatory. Rats stayed in their home cage for about 7 weeks to allow for recovery and retrograde transport of the virus. Three rats died while under anesthesia, leaving 51 rats (94%) which successfully recovered from surgery.

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Russo A.S, & Parsons R.G. (2022). Neural activity in afferent projections to the infralimbic cortex is associated with individual differences in the recall of fear extinction. Scientific Reports, 12, 13703.

Publication 2022

stereotaxic apparatus AAVrg-CAG-GFP

Anesthesia Cannula Infusion pump Ketamine Meloxicam Rats Surgery Virus Xylazine

Corresponding Organization :

Other organizations : Stony Brook University

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Variable analysis

independent variables

Injection of AAVrg-CAG-GFP virus into the IL (left- and right-side injections were counterbalanced)

dependent variables

Not explicitly mentioned

control variables

Rats were handled for two days prior to surgery
Rats were anaesthetized with ketamine (87 mg/kg) and xylazine (10 mg/kg)
Rats were placed in a stereotaxic apparatus (Stoelting, Woodale, IL)
A 22-gauge cannula was lowered into place (AP: + 3.00, ML: ± 0.6, DV: − 5.2)
A 28-gauge internal cannula (connected to an infusion pump via PE 20 tubing) was used to deliver 0.6 μL of the virus at a rate of 0.15 μL per minute and stayed in place for five minutes after the infusion was complete
Rats were injected with Meloxicam (1 mg/kg) after suturing
Rats stayed in their home cage for about 7 weeks to allow for recovery and retrograde transport of the virus

controls

No explicit mention of positive or negative controls

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