Fig. 7: RUVBL2 KO in the PVH reduces presynaptic boutons and alters synaptic vesicle distribution. | Nature Communications

Fig. 7: RUVBL2 KO in the PVH reduces presynaptic boutons and alters synaptic vesicle distribution.

From: Paraventricular hypothalamic RUVBL2 neurons suppress appetite by enhancing excitatory synaptic transmission in distinct neurocircuits

Fig. 7

a Representative images for IHC staining of the excitatory synapses in the ARC (n = 3 mice; scale bar, 100 μm). b Quantitation of VGluT2+ puncta in the ARC. AAV2-GFP (n = 17 profiles/3 mice), AAV2-Cre-GFP (n = 20 profiles/3 mice). c Representative images for IHC staining of the excitatory synapses in the DMH. (n = 3 mice; scale bar, 100 μm). d Quantitation of VGluT2+ puncta in the DMH. AAV2-GFP (n = 22 profiles/3 mice), AAV2-Cre-GFP (n = 23 profiles/3 mice). e Representative transmission electron microscopy (TEM) micrographs of the ARC. (n = 3 mice; scale bar, 200 nm). f Length of the active zone (AZ) in the ARC. AAV2-GFP (n = 75 profiles/3 mice), AAV2-Cre-GFP (n = 65 profiles/3 mice). g, h Quantity of synaptic vesicles (SVs) within different distances (g) and quantity of SVs within 200 nm (h) from the AZ in the ARC. AAV2-GFP (n = 75 profiles/3 mice), AAV2-Cre-GFP (n = 65 profiles/3 mice). i Representative TEM micrographs of the DMH (n = 3 mice; scale bar, 200 nm). j AZ length in the DMH. AAV2-GFP (n = 70 profiles/2 mice), AAV2-Cre-GFP (n = 66 profiles/2 mice). k,l Quantity of synaptic vesicles (SVs) within different distances (k) and quantity of SVs within 200 nm (l) from the AZ in the DMH. AAV2-GFP (n = 70 profiles/2 mice), AAV2-Cre-GFP (n = 66 profiles/2 mice). m GO enrichment analysis of the genes based on chromatin immunoprecipitation sequencing (ChIP-Seq). The top canonical pathways associated with the central nervous system are shown (p < 0.05). n, o Genes related to synaptic vesicle (n), or neuron axonogenesis (o) identified by ChIP-Seq. p, q Heatmap showing down-regulated genes related to synaptic vesicle (p), or neuron axonogenesis (q) by RNA sequencing (RNA-Seq). The combined analyses of RNA-Seq and ChIP-Seq (as in n, o) identify a number of genes related to synaptic vesicle or neuron axonogenesis that are down-regulated after RUVBL2 KO in the PVH. Two-tailed unpaired Student’s t-test for (b, d, f, h, j, l). ns, no significance. Two-way ANOVA followed by Bonferroni’s test for (g) and (k). All data are presented as means ± SEM. Source data are provided as a Source Data file.

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