Compensatory metabolic pathways during long-term selegiline treatment

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Fig. 3 Compensatory metabolic pathways during long-term selegiline treatment. Compensatory metabolic pathways during long-term selegiline treatment. (A) Schematic diagram of the metabolic pathway leading to GABA synthesis. (B) The experimental protocol for measuring mRNA levels by quantitative real-time polymerase chain reaction (qRT-PCR) in primary cultured astrocytes treated with selegiline (100 nM) and putrescine (180 μM) for 2 weeks. (C) Relative mRNA expression normalized to GAPDH (log10). (D) Relative mRNA expression of MAOB (0.88; n = 2), DAO (3.34; n = 2), GAD65 (0.88; n = 2), and GAD67 (0.80; n = 2). Expression levels of MAOB, DAO, GAD65, and GAD67 mRNAs in control naïve astrocytes were used to normalize different conditions. GAPDH mRNA levels were used to normalize the expression of each gene. (E) Immunostaining for GFAP and DAO after the oral administration of either selegiline or KDS2010 (10 mg/kg for 4 weeks) in APP/PS1 mice. Inset: Magnified images. (F) Mean intensity of DAO in GFAP-positive areas. *P < 0.05 and **P < 0.01, Kruskal-Wallis test and Dunnett’s multiple comparisons test. (G) Representative trace of GABAA receptor–mediated currents recorded from granule cells in the DG [n = 10 for WT + water; n = 8 for APP/PS1 + water; n = 8 for APP/PS1 + KDS2010 (10 mg/kg) for 4 weeks; n = 11 for APP/PS1 + selegiline (10 mg/kg) for 4 weeks; n = 9 for APP/PS1 + selegiline (10 mg/kg) for 4 weeks with 100 nM AG; both male and female mice aged 10 to 11 months were used]. The red bar indicates application of the GABAA receptor antagonist bicuculline (BIC) (20 μM). (H) Tonic GABA current generated by bicuculline. 2010, KDS2010. *P < 0.05 and **P < 0.01, Kruskal-Wallis test and Dunnett’s multiple comparisons test. Data are presented as means ± SEM. Bar graphs showing data distribution are presented in fig. S10. Jong-Hyun Park et al. Sci Adv 2019;5:eaav0316 Copyright © 2019 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).