Figure 1

Identification of TFLLR-induced anion conductance from hippocampal CA1 astrocytes. A. Schematic diagram of whole-cell patch clamp recordings from astrocytes in hippocampal CA1 stratum radiatum region for measuring Ca²⁺-induced glutamate conductance from CA1 astrocytes. TFLLR-NH₂ peptide (TFLLR) was used as PAR-1 agonist to induce increase in [Ca²⁺]_i in CA1 astrocytes. B. Representative current responses by voltage ramp command (from +100 mV to -100 mV) in hippocampal CA1 astrocyte. Arrows indicate ramp current before TFLLR treatment (black), after TFLLR treatment (red), and after washing (blue). C. Representative ramp current before TFLLR treatment (black trace), after TFLLR treatment (red trace), and after washing (blue trace). Dotted black line indicates 0 pA. D. Normalized average I-V relationships from whole-cell voltage clamp measurements of TFLLR-induced currents with various anions such as CsCl (n = 6), Cs-glutamate (n = 8), or gluconate (n = 7) in pipette solution. Experiment was performed in the presence of gap junction blocker 100 μM of carbenoxolone. E. Averaged I-V curves recorded from naïve, scrambled-shRNA- (sc-shRNA) or mBest1-shRNA (mB1-shRNA)-expressing astrocytes. Inset: the representative images showing lentiviral-shRNA and EGFP- expressing astrocytes stained with astrocyte marker dye (SR-101). SR-101 and EGFP-stained hippocampal CA1 astrocytes were patch clamped with 140 mM glutamate-containing pipette solution and TFLLR peptide was used to induce CAAC current. F. Bar graph representing averaged current amplitude at V_h = 80 mV or - 100 mV. Numbers on each bar indicate number of astrocytes from hippocampal slices of at least two mice. *p < 0.05.