Fig. 1: Astrocytic DRD2 facilitates GSH synthesis via Nrf2 activation. | Nature Communications

Fig. 1: Astrocytic DRD2 facilitates GSH synthesis via Nrf2 activation.

From: Pyridoxine induces glutathione synthesis via PKM2-mediated Nrf2 transactivation and confers neuroprotection

Fig. 1

a GSH levels in astrocytes and neurons after treatment with 0, 5, 10, 20, 40, or 80 µM cabergoline for 24 h. b GSH levels in astrocytes and neurons after treatment with 10 µM cabergoline for 6, 12, 18, or 24 h. c GSH levels in astrocytes treated with 10 μM cabergoline for 12 h after pretreatment with SCH23390 (10 μM) or sulpiride (10 μM). d GSH levels in astrocytes from wild-type mice or Drd2-knockout mice after quinpirole (10 μM), quinelorane (20 μM), or bromocriptine (40 μM) treatment for 12 h. Six independent experiments per condition in a–d. e Striatal GSH levels in Drd2flox/flox mice and Drd2hGFAPcKO mice after 5 mg kg−1 quinpirole administration for 7 days. n = 6 mice per group. f Representative dot plots of ACSA-2 labeling of astrocytes (gated in blue circles) collected after microbead kit separation. g, h Gene set enrichment analysis of upregulated pathways (g) and heat map of the expression of Nrf2 targets (h) identified by RNA sequencing in astrocytes from mice after quinpirole administration. i Astrocytes were treated with 10 μM quinpirole, and Nrf2 target expression was determined by qRT-PCR. Three independent experiments per condition. j, k Immunoblotting with actin as a loading control (j) and GSH levels (k) in wild-type and Nrf2-null astrocytes after 10 μM quinpirole treatment. Immunoblotting: three independent experiments; GSH assay: six independent experiments. l Diagram of the Gclc and Gclm promoters showing the locations of the different fragments tested. The numbers indicate the base pairs upstream of the transcriptional start site. m, n ChIP assays showing enrichment of the Gclc (m) and Gclm (n) promoters in DNA isolated from astrocytes treated with 10 μM quinpirole and precipitated with anti-Nrf2 antibodies. Three independent experiments per condition. Data are presented as the mean ± s.e.m. *P < 0.05, **P < 0.01, ***P < 0.001, NS, not significant. One-way ANOVA with Tukey’s multiple comparisons test (c), two-way ANOVA with Dunnett’s multiple comparisons test (a, b and d) or with Sidak’s multiple comparisons test (e, k), Student’s two-tailed unpaired t-test (i, m and n). Source data are provided as a Source Data file.

Back to article page