Fig 1: Hv1 KO mice exhibit attenuated MPTP-induced pro-inflammatory cytokine production. (A) Quantitative PCR results demonstrate that Hv1 KO mice completely abolish the MPTP-induced mRNA levels of the inflammatory cytokines Il-1b (left), Il-6 (middle), and Tnfa (right) compared to WT mice 2 days after MPTP treatment. (B) Representative immunofluorescent images of TNFα (green) in IBA1-positive (red) cells, with DAPI nuclei stain (blue) in WT and Hv1 KO sections treated with saline or MPTP, scale bar 5 μm. (C) Semi-quantification of TNFα protein fluorescent intensity levels. n = 4–5 mice per group for all experiments. Asterisks denote statistically significant differences, ** p < 0.01, *** p < 0.001, and **** p < 0.0001, using two-way ANOVA followed by Tukey’s multiple comparisons. ‘ns’ indicates not statistically significant. Error bars denote the standard error of the mean.
Fig 2: Increased HVCN1 expression in the brain of PD patients. (A) HVCN1 expression levels are increased in the substantia nigra of PD patients (n = 39 males and 22 females) compared to age-matched controls (n = 22 males and 14 females). (B) HVCN1 expression levels in male PD patients compared to male controls from the same studies. (C) HVCN1 expression levels in female PD patients compared to female controls from the same studies. GEO numbers were normalized to 1 to generate relative expression values and allow comparison between datasets. Datasets were analyzed by Mann–Whitney non-parametric test. Asterisks denote statistically significant differences—* p < 0.05, ** p < 0.01, and ns = not statistically significant. Error bars denote the standard error of the mean.
Fig 3: Hv1 deficient mice have diminished MPTP-mediated loss in TH-positive neurons in the substantia nigra. (A,C) Representative 20× DAB immunostaining images for TH in the SN of WT and Hv1 KO mice following acute 4 × 16 mg/kg (A) or 4 × 10 mg/kg (C) MPTP treatment, scale bar 50 µm. (B,D) Stereological counts for the total number of TH-positive neurons in the SN following 4 × 16 mg/kg (B) or 4 × 10 mg/kg (D) in each genotype. n = 5 mice per group. Asterisks denote statistically significant differences—** p < 0.01, *** p < 0.001, using two-way ANOVA followed by Tukey’s multiple comparisons. ‘ns’ indicates not statistically significant. Error bars denote the standard error of the mean.
Fig 4: Hv1 KO mice exhibit reduced inflammatory response and protection of TH-positive neurons in an LPS model of PD. (A–C) Quantitative PCR mRNA levels of inflammatory factors in the striatum of WT or Hv1 KO mice following sub-chronic LPS treatment, including mRNA levels of Aif1 (A), Nos2, and Gp91phox (B), and pro-inflammatory cytokines Tnfα, Il-1β, and Il-6 (C). (D) Representative 10× immunofluorescent images for TH-positive neurons in the SN following treatment of LPS in WT and Hv1 KO mice, scale bar 100 µm. (E) Stereological counts for a total number of TH-positive neurons in the SN following subchronic LPS treatment in each genotype. n = 4–5 mice per group for all experiments. Asterisks denote statistically significant differences—* p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001, using two-way ANOVA followed by Tukey’s multiple comparisons. ‘ns’ indicates not statistically significant. Error bars denote the standard error of the mean.
Fig 5: Hv1 KO attenuates LPS-induced inflammatory response in primary mouse microglia and protects dopaminergic neurons from LPS-treated microglial conditioned media. (A) Primary microglia isolated from Hv1 KO mouse pups have reduced Hv1 mRNA levels of the inflammatory cytokines Il-1b, Il-6, and Tnfa by qPCR compared to LPS-treated WT microglia. (B) Representative immunofluorescence images for TNFα (green) in Iba1-positive cells (red), with nuclear DAPI stain (blue), scale bar 20 μm. TNFα fluorescence quantification (right). (C) Quantitative PCR gene expression results for the anti-inflammatory genes Arginase-1, Igf-1, Mrc1, Ym1, and Nrf2 in WT and Hv1 KO primary microglia following LPS treatment. (D) Cell viability of N27 rat dopaminergic neurons was measured using MTS reagent that produces a formazan product in metabolically active cells. Media conditioned from primary microglia (microglia-conditioned media or MCM) treated with or without LPS for 24 h was added to N27 cells for 24 h. n = 3 isolations for primary microglia and 3 separate passages for N27 cells. Asterisks denote statistically significant differences—* p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001, using two-way ANOVA followed by Tukey’s multiple comparisons. ‘ns’ indicates not statistically significant. Error bars denote the standard error of the mean.
Supplier Page from Abcam for Anti-HV1 antibody