Microglia Responses to Methamphetamine

Reagents were purchased from the following sources: methamphetamine hydrochloride (METH) (Sigma-Aldrich); P2X7R antibody (Alomone; Jerusalem, Israel); tyrosine hydroxylase (TH) (Abcam; Cambridge, England); and pHrodo and Calcein-AM (Life Technologies; Waltham, MA). Poly-L-lysine (PLL), lipopolysaccharide (LPS), and cytochalasin D (Cyto D) were purchased from Sigma-Aldrich. Cell viability was determined by LIVE/DEAD assay (Invitrogen) and showed that METH at 1-1000 μM concentration had no toxic effects on microglia after 48 h of exposure (Additional file 1: Figure S1). The concentration of METH (100 μM) used in the present study is similar to other published studies [19 (link), 20 (link)]. Optimal concentrations of cytochalasin D (5 μM), fractalkine (CX3CL1) (10 ng/mL), LPS (1 μg/ml), pHrodo (40 μg/ml), and Calcein (5 μM) were determined from dose- and time-dependent response studies.