Fig. 1: miR-203a-3p is upregulated in ARC neurons after CCI-ION. | Nature Communications

Fig. 1: miR-203a-3p is upregulated in ARC neurons after CCI-ION.

From: Epigenetic regulation of beta-endorphin synthesis in hypothalamic arcuate nucleus neurons modulates neuropathic pain in a rodent pain model

Fig. 1

a Escape threshold in the sham- and CCI-ION-operated groups. ***p < 0.001 versus sham, two-way ANOVA followed by Bonferroni’s test. n = 10 rats per group. b Protein expression of c-fos was analyzed in bilateral ARC regions (outlined by dashed line). Arrows show Fos+ ARC neurons. Bar charts show the quantification of Fos+ neurons. *p = 0.0197 versus sham, #p = 0.0113 versus sham, two-sided Student’s t test. n = 4 rats per group. Scale bar, 100 μm. c Heatmap indicating the increased ARC-rich miRNAs in the ARC of CCI-ION rats. Color bar (up left) indicates the scale of standardized miRNA levels. n = 6 rats per group. d qPCR analysis of sixteen conserved miRNAs in the ARC. *p < 0.05, **p < 0.01, ***p < 0.001 versus sham, two-sided Student’s t test. n = 6 rats per group. e Alterations in miR-30d-5p, miR-199a-5p, miR-203a-3p and miR-451-5p in the CSF of TN patients and aged-matched healthy subjects. **p = 0.0031 versus control, two-sided Student’s t test. n = 7 subjects in control group and n = 9 subjects in TN group. f qPCR analysis of miR-203a-3p (miR-203a) expression in different brain areas, including the arcuate nucleus (ARC), anterior cingulate cortex (ACC), amygdala (AMY), dorsal raphe nucleus (DR), hippocampus (HIP), intralaminar nucleus (ILN), locus coeruleus (LC), nucleus accumbens (NAc), nucleus of the solitary tract (NTS), periaqueductal gray (PAG), parabrachial nucleus (PBN), paraventricular nucleus (PVN), primary somatosensory cortex (S1), substantia nigra (SN), supraoptic nucleus (SON), and trigeminal nucleus caudalis (TNC). Data were normalized to miR-203a-3p expression in the ARC. n = 5 rats per group. g qPCR analysis of miR-203a level in the ARC, NAc, and HIP. ***p = 0.0001 versus sham, two-sided Student’s t test. n = 8 rats per group. h–i RT‒PCR (h) and qPCR analysis (i) of miR-203a-3p expression in the ARC and TG of intact rats. ***p < 0.001 versus TG group, two-sided Student’s t test. n = 6 rats per group. j Time course of miR-203a-3p expression. ***p < 0.001 versus sham, one-way ANOVA followed by Bonferroni’s test. n = 8 rats per time point per group. k Representative FISH images of miR-203a-3p expression in the bilateral ARC. n = 3 rats per group. Scale bar, 25 μm. l FISH analysis of miR-203a-3p (red) combined with immunostaining for NeuN, GFAP, or CD11b (green). Bar charts show the percentage of double-stained ARC neurons among total numbers of miR-203a-3p+-labeled neurons. Arrows show colocalization. n = 4 rats per group. Scale bar, 25 μm. All data are presented as mean values ± SEM.

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