Adult (8–10 weeks; 200–250 g) male Sprague Dawley rats were group-housed until surgery. As previously described [60 (link)], stereotaxic-guided virus injection surgeries were performed under isoflurane anesthesia (1.5–2.5%) and a mixture of 30% N2O and 70% O2 using a stereotaxic instrument (Kopf Instruments, Tujunga, CA). Four small holes (two per side) were drilled through the skull and bilateral virus injections were made using a glass injection pipette (Drummond Scientific Company). To activate the DH, we injected AAV9-SYN-PSAM-L141F-Y115F-5HT3HC-GFP (titer: 2.19× 1013 genomes/ml, Virovek Inc.) bilaterally using the following coordinates (bregma −3 mm; lateral ±1.7 mm; ventral −3.3 mm for the more rostral location; bregma −4.2 mm; lateral ±2.7; ventral −3.3 mm for the more caudal location; 500 nl/side) at a speed of 100nl per minute. Head-posts were implanted 2–3 weeks after virus injection. Following a 2-week acclimation period, SNI surgeries were performed and 4–5 days after SNI surgery, paw withdrawal thresholds were assessed before and 0.5–2 hours post i.p. injection of either saline or PSEM89s (30 mg/kg). After completion of the experiments the injection sites were verified by immunohistochemistry.
Glass injection pipette
Manufactured by Drummond
The glass injection pipette is a laboratory tool used for accurately measuring and transferring small volumes of liquid. It consists of a thin, calibrated glass tube with a tapered tip, allowing for precise liquid manipulation and delivery.
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2 protocols using glass injection pipette
1
Optogenetic Activation of Dorsal Hippocampus in Neuropathic Pain
2
Activation of Dorsal Hippocampus in Neuropathic Pain
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Adult (8-10 weeks; 200-250 g) male Sprague-Dawley rats were group-housed until surgery. As previously described,60 (link) stereotaxic-guided virus injection surgeries were performed under isoflurane anesthesia (1.5%-2.5%) and a mixture of 30% N2O and 70% O2 using a stereotaxic instrument (Kopf Instruments). Four small holes (2 per side) were drilled through the skull, and bilateral virus injections were made using a glass injection pipette (Drummond Scientific Company). To activate the DH, we injected AAV9-SYN-PSAM-L141F-Y115F-5HT3HC-GFP (PSAM, titer: 2.19 × 1013 genomes/mL, Virovek IncHayward, CA, USA) bilaterally using the following coordinates (bregma −3 mm; lateral ±1.7 mm; ventral −3.3 mm for the more rostral location; bregma −4.2 mm; lateral ±2.7; ventral −3.3 mm for the more caudal location; 500 nL/side) at a speed of 100 nL per minute. Head posts were implanted 2 to 3 weeks after virus injection. After a 2-week acclimation period, SNI surgeries were performed, and 4 to 5 days after SNI surgery, paw withdrawal thresholds were assessed before and 0.5 to 2 hours after i.p. injection of either saline or PSEM89s (30 mg/kg). After completion of the experiments, the injection sites were verified by immunohistochemistry.
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