Journal: Neuroscience Bulletin

Article Title: ADAM17 Supports Disinhibition of Pre-sympathetic Glutamatergic Neurons Through Microglial Chemotaxis

doi: 10.1007/s12264-025-01471-8

Figure Lengend Snippet: Microglial displacement facilitates the Ang-II-induced effect of disinhibition on glutaminergic neurons in the PVN. A – C Representative images of PVN neurons and nearby cells, via transmission electron microscopy. ( A ) the microglia and neuron are illustrated by pseudo colors, red for neuron and green for microglia; Scale bar, 10 μm; ( B ) GABA stained by colloidal gold particles in microglia, as indicated in pseudo color (light blue); Scale bar, 5 μm; ( C ) GABA stained by colloidal gold particles outside of the microglia. Scale bar, 1 μm. D The AAV injection sites and a representative image of the BSTM (bed nucleus of the stria terminalis, medial division), where the neuro-tracing virus was injected. Scale bar, 50 μm. E Representative images of glutamatergic PVN-RVLM projecting neurons (PSN, green), PV + presynaptic terminals (red), and microglia (cyan) in the PVN, showing a close spatial relationship among these cells. 3D reconstruction with z-stacked images by Imaris 9. Scale bar, as indicated. F Representative immunofluorescent image of NeuN nuclei (white), Iba1 (green), and VGAT + puncta (red) in the PVN prepared 30 min following Ang-II/saline injection. Yellow arrows in the main panel and orthogonal panels indicate sites of microglial bulbous tip contact with VGAT + terminals. Scale bar, 2 μm. G Percentage of Iba1 processes colocalizing with VGAT + puncta ( n = 4 mice). Data are expressed as the mean ± SEM. (vehicle+Ang-II vs vehicle+saline, ns, no significant difference, *** P <0.001, unpaired Student’s t -test). H Representative traces of miniature IPSCs (mIPSCs) in PVN-RVLM projecting neurons from sections with Ang-II bath application and without TRAM34 pretreatment (a selective blocker of IKCa1 to inhibit microglial migration, 1 μmol/L, 30 min before Ang-II, no wash). I, J Summary of amplitude ( I ) and frequency ( J ) of mIPSCs before and after Ang-II application, in the presence of TRAM34 ( n = 5 mice/group. After vs Before, ** P <0.01, paired Student’s t -test). K Suppression ratio of Ang-II-induced reduction of mIPSC frequency, in the presence of TRAM34 (A17G vs WT, *** P <0.001, unpaired Student’s t -test; WT+TRAM34 vs WT, ††† P <0.001)

Article Snippet: In the inhibitor group, TRAM34 (Cat.# HY-13519, MCE, China) and AZD8797 (Cat.# 2255, Axon Medchem, The Netherlands) were given 30 min before Ang-II (Cat.# 4006473, BCHEM, USA).

Techniques: Transmission Assay, Electron Microscopy, Staining, Injection, Virus, Saline, Migration

Journal: Neuroscience Bulletin

Article Title: ADAM17 Supports Disinhibition of Pre-sympathetic Glutamatergic Neurons Through Microglial Chemotaxis

doi: 10.1007/s12264-025-01471-8

Figure Lengend Snippet: Pharmacologically blocking the CX3CL1/CX3CR1 chemotactic pathway blunts the Ang-II-induced pressor response. A Location of Cx3cl1 mRNA expression based on the single-nuclei RNA-seq data collected from the PVN and adjacent tissue. Upper panel: mRNA location; lower panel: UMAP for individual cell clusters. B , C Schematic and image of the administration route for Ang-II or antagonists. Scale bar, 50 μm. D Representative blood pressure (BP) recording traces from awake WT mice. E Summary data of the BP increase (mean arterial pressure, ΔMAP) in awake WT mice undergoing Ang-II injection (100 ng/100 nL) into the PVN with or without injection of AZD8797 (a selective blocker of CX3CR1, 5 μmol/L per 100 nL, 30 min before Ang-II injection); and with or without injection of TRAM34 (a selective blocker of IKCa1 to inhibit microglial migration, 10 mg/kg in 100 nL, 30 min before Ang-II injection). F Representative BP recordings from anesthetized WT mice. G Summary data of BP increase (ΔMAP) in anesthetized WT mice undergoing Ang-II injection (100 ng/100 nL) into the PVN with or without injection of AZD8797 (5 μmol/L per 100nL, 30 min before Ang-II injection); and with or without injection of TRAM34 (10 mg/kg in 100 nL, 30 min before Ang-II injection). Each data point represents an individual mouse, n = 7–9 mice. Data are shown as the mean ± SEM. * P <0.05, ** P <0.01, *** P <0.001 vs Ang-II, one-way ANOVA followed by Dunnett post tests

Article Snippet: In the inhibitor group, TRAM34 (Cat.# HY-13519, MCE, China) and AZD8797 (Cat.# 2255, Axon Medchem, The Netherlands) were given 30 min before Ang-II (Cat.# 4006473, BCHEM, USA).

Techniques: Blocking Assay, Expressing, RNA Sequencing, Injection, Migration