Journal: Nature Communications

Article Title: Radial glia integrin avb8 regulates cell autonomous microglial TGFβ1 signaling that is necessary for microglial identity

doi: 10.1038/s41467-025-57684-y

Figure Lengend Snippet: Mouse Line and Reagent List

Article Snippet: Antibody , EGFP , Origene , Goat polyclonal antibody. Cat#: R1091P. RRID:AB_1002036. , Used at 1:2000..

Techniques: RNAscope, Multiplex Assay

Journal: Cell Reports

Article Title: A cortico-subcortical loop for motor control via the pontine reticular formation

doi: 10.1016/j.celrep.2025.115230

Figure Lengend Snippet: Cortical inputs to PRF/GlyT2+ cells (A) Scheme of anterograde tracing from the frontal cortical motor-related areas (M2/Cg) in the RBP4-Cre//GlyT2-eGFP mouse line ( n = 9 mice). (B) Schematic view of the injection sites. (C) Low-power fluorescent micrographs of the cortical injection sites at three anteroposterior levels. (D) Schematic view of the M2/Cg fibers (magenta shade) and PRF/GlyT2+ cells (green dots). The black rectangle, the position of the micrographs, and heatmaps in (E)–(G). (E and F) Confocal micrographs ( n = 5 mice) of the glycinergic cells (E) and anterogradely labeled M2/Cg cortical fibers (F) in the PRF glycinergic zone. (G) Fiber density heatmap showing the distribution of cortical fibers (gray shading) and PRF/GlyT2+ cells (green dots). Higher fiber density is indicated with light gray colors. (H1–3) Maximum intensity Z projections of average cortical fiber heatmaps ( n = 3 animals) showing anterior M2/Cg inputs at three anteroposterior PRF levels extending from Br. −4.6 to Br. −4.84 (from the Paxinos atlas; 250 μm). (I) Maximum intensity Z projections of cortical fiber heatmaps ( n = 3 animals) showing posterior M2/Cg inputs. (J) Quantitative analysis of the anterior M2/Cg fibers in the PRF at three coronal levels. Intensity levels represent fiber density values between 0 and 7, where 0 indicates no innervation and 7 presents the strongest innervation. (K) Proportion of innervation densities in the PRF after anterior (left) and posterior (right) M2/Cg injections at three coronal levels. (L) High-power confocal fluorescent image of anterogradely labeled M2/Cg fibers (magenta) and PRF/GlyT2+ neurons (60×, n = 5 mice). (M–O) High-power light microscopic image ( n = 9 mice) of close apposition between M2/Cg fibers (black) and the somata (K), dendrites (L), or a spine (M) of PRF/GlyT2+ neurons. Inset in (M) displays the juxtacellularly labeled PRF/GlyT2+ cell; arrowheads, cortical inputs; asterisk, spine. Scale bars, 1 mm (C), 100 μm (E–I), 20 μm (J), 5 μm (M), 5 μm (K and L), 20 μm (inset).

Article Snippet: Chicken anti-eGFP Polyclonal Antibody , Thermo Fisher Scientific , Cat# A10262, RRID: AB_2534023 ).

Techniques: Anterograde Tracing, Injection, Labeling

Journal: Cell Reports

Article Title: A cortico-subcortical loop for motor control via the pontine reticular formation

doi: 10.1016/j.celrep.2025.115230

Figure Lengend Snippet: Cortical afferents target PRF/GlyT2+ cells and evoke a glutamatergic synaptic response (A) Scheme of anterograde tracing from the M2/Cg in the RBP4-Cre//GlyT2-eGFP mouse line. (B and C) Electron micrographs of M2/Cg terminals ( n = 44, dark precipitates) in the PRF-contacting mid-caliber dendrites. (C1–C3) Serial EM images of the same axon terminal. Black arrowheads, synapses. (D) Electron micrographs of M2/Cg terminals (DAB-Ni, dark, dense precipitate) establishing synapses on PRF/GlyT2+ dendrites ( n = 9, D1, D3, D4, DAB, light precipitate) and spines ( n = 1, D2). (E) Optogenetically evoked (blue squares) AMPA and NMDA receptor-mediated components of the EPSCs elicited by M2/Cg terminals in the PRF. Traces from one exemplary experiment are shown above. Top: AMPA and NMDA components ( n = 16), control at −60 mV ( n = 12; p = 0.002; Wilcoxon signed-rank test) and +50 mV ( n = 12; p = 0.003; Wilcoxon signed-rank test). Below, the graphs represent the results from all the experiments. (F) Top: light-evoked EPSC-s displayed no significant depression during stimulation trains, at all tested frequencies ranging from 5 to 20 Hz. Bottom: 89.1% ± 11.5% ( n = 6; p > 0.05; Wilcoxon signed-rank test), 87.3% ± 8.2% ( n = 14; p = 0.03; Wilcoxon signed-rank test), 102.7% ± 5.7% ( n = 10; p = 0.7; Wilcoxon signed-rank test), 90.5% ± 14.0% ( n = 12; p = 0.1; Wilcoxon signed-rank test at 1, 5, 10, and 20 Hz, respectively. Source data are provided as a Source Data file. Data are represented as mean ± SE. Scale bars: 1,000 nm (B), 500 nm (C), 1,000 nm (D).

Article Snippet: Chicken anti-eGFP Polyclonal Antibody , Thermo Fisher Scientific , Cat# A10262, RRID: AB_2534023 ).

Techniques: Anterograde Tracing, Control

Journal: Cell Reports

Article Title: A cortico-subcortical loop for motor control via the pontine reticular formation

doi: 10.1016/j.celrep.2025.115230

Figure Lengend Snippet: The effect of evoked cortical activity on PRF/GlyT2+ neurons (A) Scheme of the experiments ( n = 10 mice: 7 mice with optogenetic 5-ms-long pulses at 1–20 mW [RBP4-Cre//GlyT2-eGFP], 3 mice with electrical 5 ms pulses at 1–2 mA [GlyT2-eGPF]). (B) Representative stimulus-evoked cortical field activity and evoked firing of a glycinergic PRF neuron. Small ticks on the top indicate laser stimulation. (C) Representative fluorescent micrograph of a recorded and labeled PRF/GlyT2+ neuron. Green, GlyT2-eGFP; red, neurobiotin; magenta, M2/Cg fibers. (D) Peristimulus time histograms (PSTH) of cortical stimulus-evoked firing in a representative PRF/GlyT2+ neuron at 1, 10, and 20 Hz stimulation frequencies. Response probability and median latency are visualized by boxplots. 1 Hz: probability 92.86%, 12.5 ms peak ± SEM; 10 Hz: probability: 92.86%, 12.6 ms peak ± SEM, 20 Hz: probability 84.06%, 14.5 ms peak ± SEM. (E) Population PSTHs of juxtacellularly recorded and labeled PRF/GlyT2+ neurons ( n = 10) at 1, 10, and 20 Hz. 1 Hz median: 0.0136 s; 10 Hz median: 0.0127 s, 20 Hz median: 0.0132 s. (F) Median latency (left) 1 vs. 10 Hz, n.s. p = 0.22; 10 Hz vs. 20 Hz n.s. p = 0.3, Mood’s median test and response probability (right) 1 vs. 10 Hz, n.s. p = 0.59; 10 Hz vs. 20 Hz n.s. p = 0.65 Mood’s median test of the PRF/GlyT2+ neurons. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001; n.s., no significant difference. Source data are provided as a Source Data file. Scale bars, 20 μm (C).

Article Snippet: Chicken anti-eGFP Polyclonal Antibody , Thermo Fisher Scientific , Cat# A10262, RRID: AB_2534023 ).

Techniques: Activity Assay, Labeling

Journal: Cell Reports

Article Title: A cortico-subcortical loop for motor control via the pontine reticular formation

doi: 10.1016/j.celrep.2025.115230

Figure Lengend Snippet:

Article Snippet: Chicken anti-eGFP Polyclonal Antibody , Thermo Fisher Scientific , Cat# A10262, RRID: AB_2534023 ).

Techniques: Plasmid Preparation, Virus, Recombinant, Software