4 4 diisothiocyanatostilbene 2 2 disulfonic acid

The dissecting solution used for the preparation of brain slices was composed of: sucrose (252 mM), KCl (2.5 mM), CaCl₂ (0.1 mM), MgCl₂ (2 mM), Glucose (10 mM), NaHCO₃ (26 mM), and NaH₂PO₄ (1.25 mM). The extracellular fluid used for the patch-clamp recording contained: NaCl (117 mM), KCl (3.6 mM), CaCl₂ (2.5 mM), MgCl₂ (1.2 mM), NaH₂PO₄ (1.2 mM), NaHCO₃ (25 mM), and glucose (11 mM). It was continually aerated with 95% O₂ and 5% CO₂, which maintained the pH at approximately 7.4. The pipette (internal) solution contained: K-Glu (150 mM), HEPES (10 mM), KCl (5 mM), EGTA (0.1 mM), Mg-ATP (2 mM), and NaGTP (0.3 mM). The pH was adjusted to 7.2 using KOH. In low Na⁺ external solution, equimolar choline-Cl replaced NaCl. The following reagents were obtained from Sigma-Aldrich (St. Louis, MO, USA): X, XO, PBN, catalase, superoxide dismutase (SOD), 4,4-diisothiocyanatostilbene-2,2-disulfonic acid (DIDS). Dihydroethidium (DHE) and 2′,7′-dichlorofluorescin diacetate (DCF-DA) were obtained from Molecular Probes (Eugene, OR, USA).

Nicotine, RANTES, MIP-1β, 4,4′-diisothiocyanatostilbene-2,2′-disulfonic acid (DIDS), LPS, acetylcholine, bupropion, and maraviroc, were acquired from Sigma-Aldrich. Alexa-488 α-bungarotoxin was purchased from Invitrogen. gp120_JRFL and gp120_ADA were purchased from MyBioSource. gp120_IIIB was from Fitzgerald Industries International. Monoclonal α7-nAChR antibody (Cat. # sc-5544, Santa Cruz Biotechnology), monoclonal GAPDH antibody (Cat. # ab8645, Abcam), and goat anti-mouse IgG HRP (Cat. # sc-2005, Santa Cruz Biotechnology) were obtained from Santa Cruz Biotechnology. An anti-goat secondary antibody-labeled HRP-conjugated were obtained from Sigma Aldrich (Cat. # AP307P, EMD Millipore).