Cgs 21680

The preparation of the fibrin glue and the selection of the A2A receptor agonist and antagonist doses were based on the methods reported by several articles [27 (link)–30 (link)]. Briefly, fibrinogen powder (Solarbio, China) was dissolved in physiological saline into a concentration with 10 mg/ml at 37°C. Thrombin powder (Solarbio, China) was dissolved in 40 mmol/l calcium chloride solution, and the concentration was 20 μg/ml. Then, 5 mg CGS 21680 (specific adenosine A2A receptor agonist; Selleck, USA) or ZM 241385 (specific adenosine A2A receptor antagonist; Selleck, USA) was dissolved in 75 μl DMSO and mixed with thrombin solution into a concentration with 16 mg/ml CGS 21680 or ZM 241385 thrombin solution. Afterwards, 125 μl CGS 21680, ZM 241385, or thrombin solution (with DMSO as the solvent accelerating agent) was mixed with 125 μl fibrinogen solution to form a fibrin gel drug delivery system after 1 min of mixing.

All drugs were dispensed on the spot. Clozapine‐N‐oxide (CNO) (Tocris, 4936), fluorocitric acid (FC) (Sigma, F9634), DCPCX (MCE, HY‐100937), SCH58261 (Selleck, S8104), and CGS 21680 (Selleck, S2153) were prepared as stock solutions in dimethyl sulfoxide (DMSO) and subsequently diluted to their final concentrations in sterile 0.9% saline. CCPA (Sigma, C7938) was dissolved directly in sterile 0.9% saline and further diluted to the desired concentration. The final concentrations employed were 100 mM for FC and 10 mM for CCPA, DCPCX, SCH58261, and CGS21680. For intraperitoneal (i.p.) injection, rats were administered CNO at 3 mg/kg body weight or the corresponding vehicle. In the case of Ca²⁺ and GRAB_Ado imaging, 10 mM CNO was applied to the slices by dissolving it in the perfused artificial cerebrospinal fluid (ACSF). Intracerebral drug delivery was facilitated through previously implanted infusion cannulas. On the day of the experiment, the internal cannulas, extending 0.5 mm beyond the ends of the guide cannulas, were inserted, and drugs (500 nL/side) were infused. The control groups received an equivalent amount of DMSO dissolved in sterile 0.9% saline or an equivalent volume of sterile 0.9% saline.