Journal: The Journal of Neuroscience

Article Title: Dorsal–Caudal and Ventral Hippocampi Target Different Cell Populations in the Medial Frontal Cortex in Rodents

doi: 10.1523/JNEUROSCI.0217-25.2025

Figure Lengend Snippet: Anterograde transsynaptic spread of AAV1-Cre injections from dcHPC and vHPC target superficial and deep layers of MFC in mice. A , AAV1-hSyn-Cre was injected in either dcHPC or vHPC resulting in expression in HPC projection neurons (data not shown). The transsynaptic spread to postsynaptic neurons was detected by immunostaining against Cre. Note that postsynaptic neurons likely include principal neurons and interneurons in MFC which, in the scheme, are represented by triangles and circles, respectively. The present experiments do not include a further differentiation between interneurons and principal neurons. B , Summary of AAV1-Cre injection sites along HPC displayed in coronal sections. Injection sites in dcHPC and vHPC are shown in cool and warm colors, respectively. C , Representative samples of MFC coronal sections showing the distribution of postsynaptic Cre-expressing neurons in MFC for dcHPC injections (left, case #567) and high-magnification image of the boxed region (right). Scale bars, 500 µm. D , Representative samples of MFC coronal sections showing the distribution of postsynaptic Cre-expressing neurons in MFC for vHPC injections (left, case #565) and high-magnification image of boxed region (right). Scale bars, 500 µm. E , Representative scheme showing the distribution of postsynaptic Cre-labeled neurons across the anterior–posterior axis of MFC for one dcHPC (case #567, green) and one vHPC (case #565, magenta) injection. F , Proportion of the number of Cre-labeled cells in a MFC subregion among the total Cre-labeled neurons across MFC sections for dcHPC (green, n = 5) and vHPC (magenta, n = 5) injections. Data are presented as violin plots, with each circle corresponding to one sample. Values from all subregions sum up to 1 for one individual sample. Mann–Whitney test corrected for multiple comparisons using the Holm–Šídák method: * p < 0.05. G , H , Laminar distribution of postsynaptic Cre-labeled cells in superficial and deep layers of MFC subregions for dcHPC ( E ) and vHPC ( F ) injections. Data are presented as violin plots, with each circle corresponding to one sample. Values from superficial and deep layers of all subregions sum up to 1 for one individual sample. Mann–Whitney test corrected for multiple comparisons using the Holm–Šídák method: * p < 0.05.

Article Snippet: For the anterograde transsynaptic experiments, AAV1-hSyn-Cre (1.9 × 10 13 to 2.1 × 10 13 GC/ml; 200 nl; Addgene-10553) was injected in either dcHPC or vHPC, and either AAV8-hSyn-DIO-mCherry (3.6 × 10 12 GC/ml; 3 × 100 nl; Addgene-50459) or AAV9-hDlx-Flex-GFP (4.4 × 10 12 GC/ml; 3 × 100 nl; Addgene-83895) was injected along the dorsoventral axis of MFC.

Techniques: Injection, Expressing, Immunostaining, Labeling, MANN-WHITNEY

Journal: The Journal of Neuroscience

Article Title: Dorsal–Caudal and Ventral Hippocampi Target Different Cell Populations in the Medial Frontal Cortex in Rodents

doi: 10.1523/JNEUROSCI.0217-25.2025

Figure Lengend Snippet: dcHPC and vHPC inputs to MFC innervate GABAergic and non-GABAergic neurons in mice. A , AAV1-hSyn-Cre was injected in either dcHPC or vHPC and was expressed in HPC projection neurons. The transsynaptic spread led to Cre expression in postsynaptic neurons, including principal neurons and interneurons in MFC. In parallel, AAV8-hSyn-DIO-mCherry was injected in MFC leading to Cre-dependent mCherry expression in postsynaptic neurons in MFC. Postsynaptic interneurons were subsequently differentiated by immunostaining against GAD67, as illustrated in D–F . In the scheme, principal neurons and interneurons are represented by triangles and circles, respectively. B , Summary of AAV1-hSyn-Cre injection sites along the HPC displayed in coronal sections. Injection sites in the dcHPC and vHPC are shown in cool and warm colors, respectively. C , Representative samples with AAV1-hSyn-Cre injection in either dcHPC (left, case #611) or vHPC (right, case #696). Images show the postsynaptic Cre-dependent mCherry expression in a coronal section of MFC. Scale bar, 500 µm. D , Representative sample showing the mCherry expression of postsynaptic neurons and colocalization with the GAD67 antibody labeling as used to identify the GABAergic neurons among the HPC-MFC postsynaptic cells. Scale bar, 20 µm. E , Proportion of cells coexpressing mCherry and GAD67 among the total mCherry+ neurons receiving dcHPC (green, n = 6) or vHPC (pink, n = 6) inputs in MFC. The proportions were compared with the native presence of GAD67 in MFC (light-blue, n = 4). One-way ANOVA, followed by post hoc Tukey's multiple-comparison test: *** p < 0.001; **** p < 0.0001. F , Subregional distribution of GABAergic (GAD+) and non-GABAergic (GAD−) neurons in MFC receiving inputs from dcHPC (left, n = 6, green tones) and vHPC (right, n = 6, pink tones) Cre injections. The proportions of inputs to GAD+ and GAD− cells across MFC subregions sum up to 1 for each injection sample. Data are presented as violin plots, where each circle corresponds to one sample. Two-tailed t tests: ** p < 0.01; * p < 0.05.

Article Snippet: For the anterograde transsynaptic experiments, AAV1-hSyn-Cre (1.9 × 10 13 to 2.1 × 10 13 GC/ml; 200 nl; Addgene-10553) was injected in either dcHPC or vHPC, and either AAV8-hSyn-DIO-mCherry (3.6 × 10 12 GC/ml; 3 × 100 nl; Addgene-50459) or AAV9-hDlx-Flex-GFP (4.4 × 10 12 GC/ml; 3 × 100 nl; Addgene-83895) was injected along the dorsoventral axis of MFC.

Techniques: Injection, Expressing, Immunostaining, Antibody Labeling, Comparison, Two Tailed Test

Journal: The Journal of Neuroscience

Article Title: Dorsal–Caudal and Ventral Hippocampi Target Different Cell Populations in the Medial Frontal Cortex in Rodents

doi: 10.1523/JNEUROSCI.0217-25.2025

Figure Lengend Snippet: dcHPC and vHPC innervate both PV and SOM interneurons in the mouse MFC. A , Representative images showing the native distribution of the indicated marker expression (left, PV; right, SOM) in the subregions of MFC. Scale bar, 500 µm. B , AAV1-hSyn-Cre was injected in either dcHPC or vHPC and expressed in HPC projection neurons. The transsynaptic spread led to Cre expression in postsynaptic neurons, including principal neurons and interneurons in MFC. In parallel, AAV9-hDlx-Flex-GFP was injected in MFC, leading to Cre-dependent GFP expression in postsynaptic interneurons in MFC. In the scheme, principal neurons and interneurons are represented by triangles and circles, respectively. C , Summary of AAV1-hSyn-Cre injection sites along HPC in coronal sections. Injection sites in dcHPC and vHPC are shown in cool and warm colors, respectively. D , Micrographs showing the viral GFP expression of labeled GABAergic neurons and colocalization with the GAD67 antibody. Scale bar, 20 µm. E , Micrographs showing the viral GFP expression and colocalization of GFP+ cells and the indicated markers (PV+ in red, SOM+ in violet). Scale bar, 100 µm. F , Representative schemes showing the distribution of the viral GFP-expressing interneurons in MFC receiving inputs from dcHPC (left, case #773) and vHPC (right, case #772) Cre injections that coexpress either PV or SOM across the MFC subregions. G , Pie charts showing the proportion of GFP+ cells coexpressing PV (red) or SOM (violet) among the total GFP-labeled interneurons receiving dcHPC (left, n = 6) or vHPC (right, n = 4) inputs. H , I , Subregional distribution of the GFP+ interneurons receiving HPC inputs in MFC that coexpress the indicated markers PV (red) or SOM (violet) for dcHPC ( H , n = 6) and vHPC ( I , n = 4). Data are presented as violin plots, with each circle corresponding to one sample. For one sample, proportions of PV+ and SOM+ cells across the MFC subregions do not sum up to 1, with the residual proportion corresponding to unidentified GFP+ interneurons. A Mann–Whitney test corrected for multiple comparisons using the Holm–Šídák method.

Article Snippet: For the anterograde transsynaptic experiments, AAV1-hSyn-Cre (1.9 × 10 13 to 2.1 × 10 13 GC/ml; 200 nl; Addgene-10553) was injected in either dcHPC or vHPC, and either AAV8-hSyn-DIO-mCherry (3.6 × 10 12 GC/ml; 3 × 100 nl; Addgene-50459) or AAV9-hDlx-Flex-GFP (4.4 × 10 12 GC/ml; 3 × 100 nl; Addgene-83895) was injected along the dorsoventral axis of MFC.

Techniques: Marker, Expressing, Injection, Labeling, MANN-WHITNEY

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: The other two cases we used BAC_glyt2/gfp mouse line ( n = 2) and we injected AAVrg-EF1a-Cre (RRID:Addgene_55636) into the Pf unilaterally (AP: −2, ML: 0,5, DV: 3; 30 nL).

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: The other two cases we used BAC_glyt2/gfp mouse line ( n = 2) and we injected AAVrg-EF1a-Cre (RRID:Addgene_55636) into the Pf unilaterally (AP: −2, ML: 0,5, DV: 3; 30 nL).

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: The other two cases we used BAC_glyt2/gfp mouse line ( n = 2) and we injected AAVrg-EF1a-Cre (RRID:Addgene_55636) into the Pf unilaterally (AP: −2, ML: 0,5, DV: 3; 30 nL).

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: Co-innervation of PRF and thalamus by M2/Cg L5 neurons (A) Reconstruction of a representative M2 L5 (AA0245) neuron from the Mouse Light Neuron Browser database which innervates both PRF and IL/Pf (top). Axon arbor of the representative M2 L5 neuron (AA0245) at the sagittal level in the PRF (bottom). (B) Axon arbor of the representative M2 L5 neuron (AA0245) in the IL (top) and Pf (bottom). (C) Axon arbor of the representative M2 L5 neuron (AA0245) (left in red) and of 4 L5 neurons in the PRF (right, red, yellow, blue, and green) that have 10 or more axonal endpoints in the PRF and also innervate IL/Pf with multiple endpoints. (D) Scheme of the viral tracing to label PRF collaterals of thalamus-projecting M2/Cg cells. (E) Low-power confocal micrograph of the cortical M2/Cg injection site. (F) High-power confocal fluorescent image of the thalamus-projecting M2 L5 neurons. (G) Merged confocal image of the labeled thalamus-projecting M2/Cg cortical fibers (magenta) and PRF/GlyT2+ fibers (green) in the thalamus. (H) Merged, low-power confocal image of the labeled thalamus-projecting M2/Cg cortical fibers (magenta) and PRF/GlyT2+ fibers (green) in the brainstem. (I–K) Confocal micrographs of the PRF/GlyT2+ cells (green, I), anterogradely labeled thalamus-projecting M2/Cg cortical fibers (magenta, J), and their merged image (K). (L–N) High-power confocal microscopic image of close apposition between the PRF/GlyT2+ dendrites (green, L) and thalamus-projecting M2/Cg fibers (magenta, M). Arrows, putative contacts in (N). (O) Percentage of innervated PRF/GlyT2+ dendrite (left) and the mean number of the close apposition per dendritic segment (right). Source data are provided as a Source Data file. Scale bars, 1 mm (E), 500 μm (F and G), 1 mm (H), 100 μm (I–K), 5 μm (L–N).

Article Snippet: The other two cases we used BAC_glyt2/gfp mouse line ( n = 2) and we injected AAVrg-EF1a-Cre (RRID:Addgene_55636) into the Pf unilaterally (AP: −2, ML: 0,5, DV: 3; 30 nL).

Techniques: 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: Inhibitory action and cortical innervation of thalamus-projecting PRF/GlyT2+ cells (A) Top: scheme of the juxtacellular recordings ( n = 13 mice, n = 23 neurons). Bottom: post hoc identified IL/Pf neuron surrounded by GlyT2+ fibers. (B) Response of two representative IL/Pf thalamic neurons to optogenetic activation of PRF/GlyT2+ fibers (top, 5 s stimulation, without a tail pinch; bottom, 30 s stimulation with a tail pinch). Top: discriminated action potentials (AP), 5 s stimulation, without tail pinch. Bottom: instantaneous firing rate (blue line, laser ON), 30 s stimulation with a tail pinch. (C) Mean firing rate (MFR) of the recorded IL/Pf thalamic neurons before, during, and after the photoactivation PRF/GlyT2+ fibers (33 Hz, 5 ms pulse width, laser power 10 mW, 5–30 s duration). Strongly inhibited, (red circles, n = 19 cell), weakly inhibited (light blue circles, n = 4, see STAR Methods) and neurons responding with a post-stimulus gap (dark blue circle, n = 4) are indicated. No pinch (left) and pinch (right) conditions are shown separately. No pinch MFR before vs. stim p = 0.002; no pinch MFR stim vs. after p = 0.002, n = 10 cells; pinch before vs. stim p = 0.00012; pinch MFR stim vs. after p = 0.00085, n = 14 cells; Wilcoxon signed-rank test. (D) Scheme of the double conditional viral tracing to label cortical inputs to thalamus-projecting PRF/GlyT2+ cells. (E) Low-power confocal micrograph of the cortical injection site. (F and G) Merged confocal image of the M2/Cg cortical fibers (green) and PRF/GlyT2+ fibers (magenta) in the thalamus. The red rectangle in (F) indicates the position of the micrograph in (G). (H) Coronal, stereotactic image of the brainstem. The red rectangle indicates the position of the micrographs in (I)–(K). (I–K) Confocal micrographs of the anterogradely labeled M2/Cg cortical fibers (green, I), retrogradely labeled thalamus-projecting PRF/GlyT2+ cells (magenta, J), and their merged image (K). (L–N) High-power confocal microscopic image of the thalamus-projecting PRF/GlyT2+ dendrites (magenta, L) and M2/Cg fibers (magenta, M) and their putative contacts (N, arrowheads). (O) Percentage of innervated thalamus-projecting PRF/GlyT2+ dendrite (left) and mean number of the close apposition per dendritic segment (right). ∗ 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 (A), 1 mm (E–G), 50 μm (I–K), 5 μm (L–N).

Article Snippet: The other two cases we used BAC_glyt2/gfp mouse line ( n = 2) and we injected AAVrg-EF1a-Cre (RRID:Addgene_55636) into the Pf unilaterally (AP: −2, ML: 0,5, DV: 3; 30 nL).

Techniques: Activation Assay, 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: Rotational movements evoked by axonal vs. somatic photoactivation of PRF/GlyT2+ neurons (A) Experimental design of the photoactivation of thalamus-projecting PRF/GlyT2+ cells via their fibers in the thalamus ( n = 7 fiber optic fibers). (B) Contralateral rotations ( n = 7 fiber optics) displayed as mean rotation angle before, during, and after the stimulus periods following PRF/GlyT2+ fiber stimulation in Pf. The figure represents the average percentage of a full 360° circle per second (χ 2 = 10.6, p = 0.005, Friedman test following Durbin-Conover post hoc, before vs. stim p < 0.001; stim vs. after p < 0.001). (C) Schematic view of post hoc identified 7 optic fiber positions for experimental animals in Pf. (D) Experimental design of the photoactivation of PRF/GlyT2+ somata in the PRF ( n = 7 fiber optics). (E) Movement trajectory in one representative mouse displaying contralateral turning (mouse ID: 1). Non-stimulated period (left), stimulated period (right) at 5 mW in the PRF. (F) Comparison of the absolute values of the mean rotation angle before, during, and after the PRF stimulus periods in the 7 experimental mice (Friedman test following Durbin-Conover post hoc; 1 mW χ 2 = 6, p = 0.050; before vs. stim p = 0.073; stim vs. after p = 0.012; 5 mW χ 2 = 8.33, p = 0.016; before vs. stim p = 0.038; stim vs. after p < 0.001; 10 mW χ 2 = 10.3, p = 0.006; before vs. stim p < 0.001; stim vs. after p = 0.014; 15 mW χ 2 = 12.3, p = 0.002; before vs. stim p=<0.001; stim vs. after p = 0.004 5 mW, χ 2 = 8.333, p = 0.012. (G) Lack of significant dose-response effect on rotation (Friedman test χ 2 = 7, p = 0.072). (H) Contralateral ( n = 4 animals, blueish shades) and ipsilateral ( n = 3 animals, reddish shades) rotations following PRF/GlyT2 somatic stimulations. The color code of the animals is the same as in (F). (I) Schematic view of post hoc identified optic fiber position ( n = 7) in PRF for the 7 experimental mice. Right, fiber optic positions inducing contralateral rotation; left, fiber optic positions inducing ipsilateral rotation. Same notation as in (G). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001; n.s., no significant difference. Source data are provided as a Source Data file.

Article Snippet: The other two cases we used BAC_glyt2/gfp mouse line ( n = 2) and we injected AAVrg-EF1a-Cre (RRID:Addgene_55636) into the Pf unilaterally (AP: −2, ML: 0,5, DV: 3; 30 nL).

Techniques: Comparison

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: Selective ipsilateral innervation of the gigantocellular nucleus by thalamus-projecting PRF/GlyT2+ cells (A) (A1) Experimental design to label thalamus-projecting PRF/GlyT2+ cells+ and their axons ( n = 3 mice). (A2–3) Fluorescent micrograph about virus injection site in the PRF. The red rectangle on (A2) indicates the position of the micrograph in (A3). (A4) Ipsilateral projection of the thalamus-projecting PRF/GlyT2+ cells in the Gi (yellow). (A5) Quantification of ipsilateral and contralateral axonal projections of the thalamus-projecting PRF/GlyT2+ cells in the Gi in GlyT2-Cre mice (paired t test, t (2) = 22.07, p = 0.002, n = 3 mice). (B) (B1) Experimental design to label all PRF/GlyT2+ cells ( n = 4 mice). (B2–3) Fluorescent micrograph about virus injection site in PRF. The red rectangle on (B2) indicates the position of the micrograph in (B3). (B4) Bilateral projection of the PRF/GlyT2+ cells in the Gi (green). (B5) Quantification of the ipsilateral vs. contralateral axonal projections from PRF/GlyT2+ cells within the Gi region in GlyT2-Cre mice (paired t test, t (3) = 1.462, p = 0.24, n = 4 mice). (C) (C1) Scheme of mixed viral injection into PRF in vGAT-Flp/vGlut2-Cre ( n = 3 mice). (C2–3) Fluorescent micrograph about virus injection site in PRF. The red rectangle on C2 indicates the position of the C3 micrograph. (C4) Bilateral projection with contralateral dominance of PRF/vGlut2+ cells in the Gi (magenta). (C5) Quantification of ipsilateral and contralateral axonal projections from PRF/vGluT2+ cells within the Gi region in vGluT2-Cre/vGAT-FLP mice (paired t test, t (2) = 19.04, p = 0.0027, n = 3 mice). (D) (D1–2) Fluorescent micrograph about virus injection site in PRF. The red rectangle on (D1) indicates the position of the micrograph in (D2). (D3) Bilateral projection with ipsilateral dominance of PRF/vGAT+ cells in the Gi (cyan). (D4) Quantification of ipsilateral and contralateral axonal projections from PRF/vGAT2+ cells within the Gi region in vGluT2-Cre/vGAT-Flp mice (paired t test, t (2) = 5.61, p = 0.0303, n = 3 mice). (E) (E1–2) Fluorescent micrograph about virus injection site in PRF. The red rectangle on (E1) indicates the position of the micrograph in (E2). (E3) Merged image of PRF/vGAT+ (cyan) and PRF/vGlut2+ (magenta) fibers in the Gi. ∗ 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, 500 μm (A2, B2, C2, D1, E1), 100 μm (A3, B3, C3, D2, E2), 500 μm (A4, B4, C4, D3, E3).

Article Snippet: The other two cases we used BAC_glyt2/gfp mouse line ( n = 2) and we injected AAVrg-EF1a-Cre (RRID:Addgene_55636) into the Pf unilaterally (AP: −2, ML: 0,5, DV: 3; 30 nL).

Techniques: Virus, Injection

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: The other two cases we used BAC_glyt2/gfp mouse line ( n = 2) and we injected AAVrg-EF1a-Cre (RRID:Addgene_55636) into the Pf unilaterally (AP: −2, ML: 0,5, DV: 3; 30 nL).

Techniques: Plasmid Preparation, Virus, Recombinant, Software