Intracerebroventricular Cytokine Delivery

Delivery of TNF-α, IL-1β ± HB-EGF by i.c.v. injection was performed as described previously^{18 (link),20 (link),53 (link)}. In brief, mice were anesthetized using 1% isoflurane mixed with oxygen. Heads were shaved and cleaned using 70% ethanol and lidocain gel followed by a medial incision of the skin to expose the skull. The ventricles were targeted bilaterally using the coordinates: ±1.0 (lateral), −0.44 (posterior), −2.2 (ventral) relative to Bregma. Mice were injected with 10 μl of vehicle or cytokine solution containing 100 ng of TNF-α (Peprotech, no. AF-315-01A), IL-1β (Peprotech, no. 211-11B) ± HB-EGF (Novus Biologicals, no. 35069) using a 10-μl Hamilton syringe (Sigma Aldrich, no. 20787) on a Stereotaxic Alignment System (Kopf, no. 1900), sutured and permitted to recover in a separate clean cage. Mice received a subcutaneous injection of 1 mg kg⁻¹ meloxicam post i.c.v. injection and were analyzed after 24 h.