Both male and female adult (8–12 weeks old) Thy1-yellow fluorescent protein mice, line H (Thy1-YFP-H; Jackson Laboratories, Bar Harbor, USA) [43 (link)], were used in these studies. All experimental procedures were approved by the IPMon Animal Care Committee and conducted according to international FELASA guidelines, national law, and ethical guidelines (Uruguayan Animal Care Committee).
The surgical procedure was carried out following sterile precautions. Mice were anesthetized with ketamine-xylazine (90–10 mg/kg) and the right sciatic nerve at the mid thigh level was exposed. Treatments were performed by direct injections at 45 mm from the tip of the third digit in 2 μl of sterile PBS, using a fine glass micropipette connected to a Hamilton syringe. Immediately after the injection and at the same position, the nerve was crushed in two different directions 30 s each time with fine forceps. The crush site was labeled with lamp black powder. The wound was closed with 5–0 mononylon Ethilon sutures (Ethicon) and disinfected. The sciatic and tibial nerves and the plantar skin were harvested at 24 h, 3, 10, and 28 days post lesion (dpl).
All nerve injections were performed in 2 μl PBS and at 10 μg/ml concentration of the following products: rCD300f-IgG2a (rat extracellular domain of CD300f fused to mouse IgG2a protein) or purified mouse myeloma IgG2a (Invitrogen, Cat. N° 026200).
The surgical procedure was carried out following sterile precautions. Mice were anesthetized with ketamine-xylazine (90–10 mg/kg) and the right sciatic nerve at the mid thigh level was exposed. Treatments were performed by direct injections at 45 mm from the tip of the third digit in 2 μl of sterile PBS, using a fine glass micropipette connected to a Hamilton syringe. Immediately after the injection and at the same position, the nerve was crushed in two different directions 30 s each time with fine forceps. The crush site was labeled with lamp black powder. The wound was closed with 5–0 mononylon Ethilon sutures (Ethicon) and disinfected. The sciatic and tibial nerves and the plantar skin were harvested at 24 h, 3, 10, and 28 days post lesion (dpl).
All nerve injections were performed in 2 μl PBS and at 10 μg/ml concentration of the following products: rCD300f-IgG2a (rat extracellular domain of CD300f fused to mouse IgG2a protein) or purified mouse myeloma IgG2a (Invitrogen, Cat. N° 026200).