FIG. 2.

Difference in ROS management between pyramidal cells in CA1 and CA3 is evidenced by artificially stimulating the generation and removal of ROS. (A) (1) Superoxide is generated by inhibiting complex I with MD; (2) superoxide dismutation can be accelerated with MnSOD_m. MnSOD_m at 20 μM was found to be sufficient to fully convert superoxide into H₂O₂ (see Supplementary Fig. S4 for more details). (3) H₂O₂ is removed by the GSH and Trx systems. In (B–E), there are five stages (labeled with I to V) in the treatment. Each trace is represented as mean ± SEM, n = 6 OHSCs. (B) OxD_P of the H₂O₂ probe is monitored in CA1 and CA3 during 5 min of basal conditions (I), 15 min; 10 μM MD (II), 15 min; WO (III); followed by 15 min 10 μM MD +20 μM MnSOD_m (IV); and finally15 min WO (V). ACSF is used for basal and WO. One-way ANOVA test results of CA1 and CA3 are indicated by black and red horizontal lines, respectively. Student's t-test results are indicated by vertical lines. (C) OxD_P is monitored in CA1 and CA3 during the treatment as above, but with higher MD concentration. (D) OxD_G of GSH probe is monitored in CA1 and CA3 when OHSC is treated in the same condition as in (B). (E) OxD_G is monitored in CA1 and CA3 treated in the same conditions as in (C). See Figure 1 legend for indicators of statistical significance. ACSF, artificial cerebrospinal fluid; MD, menadione; ROS, reactive oxygen species; WO, washout.