Journal: bioRxiv

Article Title: Coordination of distinct sources of excitatory inputs enhances motion selectivity in the mouse visual thalamus

doi: 10.1101/2025.01.08.631826

Figure Lengend Snippet: (A) Schematic of potential outcomes on direction-selective (DS) dLGN neurons after silencing collicular input. Displacement, shift of the preferred motion direction; subtractive, divisive, and selective suppression are defined in (B). (B) Schematic illustrating direction tuning curves of DS dLGN neurons under three types of suppression after silencing collicular input. Subtractive: Responses are shifted down equally across directions, with the suppression ratio peaking in the null direction. Divisive: Responses are suppressed by the same ratio across all directions. Selective: Responses are suppressed by a higher ratio in the preferred direction. (C) Scatterplot of the suppression ratio in the null direction vs. the preferred direction for DS dLGN neurons under subtractive (orange), divisive (blue), or selective (green) suppression, or enhancement following collicular input silencing. Results in C, D, G-O are from 27 FOVs of 11 hM4Di-injected Rorβ-Cre mice. (D) Distinct distributions of the ratio between the suppression ratios in the null direction and the preferred direction for dLGN neurons under distinct types of suppression in C. (E) Larger fractions of DS dLGN neurons underwent divisive and selective suppression in (E 1 ) hM4Di-injected Rorβ-Cre mice (n = 17, 70, 64, and 23 neurons for subtractive, divisive, selective suppression, and enhanced categories, respectively) compared to (E 2 ) mCherry-injected Rorβ-Cre mice (n = 13, 40, 28, and 69 for subtractive, divisive, selective suppression, and enhanced categories). p<0.001, Chi-squared test; p = 0.73, 0.010, 0.0003, and 8.9*10 -10 for subtractive, divisive, selective, enhanced categories, respectively, post hoc Pearson residuals test. (F) (F 1 ) hM4Di-injected Vglut2 mice had a significantly higher proportion of selectively-suppressed excitatory DS neurons (n = 10, 70, 78 and 12 for subtractive, divisive, selective, and enhanced categories, respectively) compared to (F 2 ) mCherry-injected Vglut2 mice (n = 4, 62, 44 and 51 for subtractive, divisive, selective, and enhanced categories, respectively). p<0.001, Chi-squared test; p = 0.12, 0.62, 0.0004, and 1.4*10 -7 for subtractive, divisive, selective, and enhanced categories, respectively, post hoc Pearson residuals test. (G) Averaged tuning curves after saline injection and after CNO injection for the DS dLGN neurons that exhibited subtractive suppression following CNO injection (p = 0.04 at -180°, p = 0.02 at 0°; n = 17 neurons). Peak directions are set to 0°. (H) Suppression ratios across all normalized directions for neurons in G (p = 0.14, Kruskal-Wallis test). (I) Significantly different DSI after CNO injection compared to saline injection in subtractively-suppressed DS neurons (DSI: after saline inj. 0.326 ± 0.044, after CNO inj. 0.452 ± 0.056; p < 10 -3 ). (J) Averaged tuning curves after saline injection and after CNO injection for the DS dLGN neurons that exhibited divisive suppression following CNO injection (p = 0.012 at -180°, p < 10 -4 at 0°; n = 70 neurons). (K) Suppression ratios across all normalized directions were not significantly different for neurons in J (p = 0.056; Kruskal-Wallis test). (L) DSI values after CNO injection and after saline injection were not significantly different in divisively-suppressed DS dLGN neurons (DSI: after saline inj. 0.448 ± 0.024, after CNO inj. 0.453 ± 0.024; p = 0.90). (M) Similar to (G) but for the DS dLGN neurons exhibiting selective suppression for the preferred direction after CNO injection (p = 0.54 at -180°, p < 0.001 at 0°; n = 64 neurons). (N) Suppression ratios for neurons in M were significantly different across normalized directions (p < 0.001, Kruskal-Wallis test; p = 0.0006 between -180° and 0°, p = 0.02 between -135° and 0°, p = 0.02 between -90° and 0°, p = 0.02 between -45° and 0°, p = 0.011 between 0° and 135°, Dunn’s multiple comparisons test). (O) In selectively-suppressed DS dLGN neurons, the DSI after CNO injection was significantly lower than after saline injection (DSI: after saline inj. 0.405 ± 0.024, after CNO inj. 0.352 ± 0.024. p = 0.04). All the data was plotted as mean ± SEM. *, p < 0.05; **, p < 0.01; ***, p < 0.001. G, I, J, L, M and O: linear mixed-effects model. See also , .

Article Snippet: The primary antibodies used were chicken anti-GFP (1:1000, Invitrogen, A10262; used for GCaMP6f), rabbit anti-RFP (1:1000, Rockland, 600-401-379, used for jRGECO1a and mCherry), guinea pig anti-RBPMS (1:500, PhosphoSolutions, 1832-RBPMS), guinea pig anti-VGLUT2 (1:500, Synaptic systems, 135404), and mouse anti-VGAT (1:200, Synaptic systems, 131011).

Techniques: Injection, Saline

Journal: Acta Neuropathologica Communications

Article Title: Multiparametric grading of glaucoma severity by histopathology can enable post-mortem substratification of disease state

doi: 10.1186/s40478-024-01880-2

Figure Lengend Snippet: Multivariate statistical analysis. A Principal component analysis for GFAP, RBPMS, NEFL and IBA1 immunoreactivity demonstrated that > 85% of the variance in the data was contributed by the first 2 principal components, as indicated in the Scree plot. B Score plot from factor analysis showing that maximal variance was weighted by 3 factors: RBPMS, NEFL and IBA1, with the former 2 representing neuronal components, and the latter representing a glial component. C Cluster analysis using the principal components. Severity of glaucoma was assessed by applying the Calinski-Harabasz criterion for unbiased k-means clustering using the expression values for RBPMS, NEFL, and GFAP positive staining. Non-glaucomatous donors clustered in 2 distinct groups while glaucomatous tissues clustered in 3 distinct groups (mild, moderate, and severe). Arrows as indicated in the figure identify tissues with qualitative cupping of the ONH observed through histology

Article Snippet: IHC was performed with antibodies (Table ) against RBPMS (RGC soma marker), NEFL (RGC axon marker), GFAP (astrocyte marker) and IBA1 (microglia/macrophage marker) using a LeicaBond RX stainer (Leica Biosystems, Deer Park, IL, USA).

Techniques: Expressing, Staining

Journal: Acta Neuropathologica Communications

Article Title: Multiparametric grading of glaucoma severity by histopathology can enable post-mortem substratification of disease state

doi: 10.1186/s40478-024-01880-2

Figure Lengend Snippet: RGC loss in glaucomatous donors observed by loss of RBPMS positive cells. A Loss of RGCs in peripheral retina and macula was observed in glaucomatous donors; arrows indicate RBPMS positive cells. B Scatter dot plot represents the individual values of RBPMS-positive RGCs (RBPMS + RGCs) for non-glaucomatous (n = 16) and glaucomatous donor tissues (n = 21). Quantitative analysis demonstrated a 54% reduction in RBPMS + RGCs in glaucomatous temporal peripheral retina. Horizontal bars indicate mean ± standard deviation. ****p < 0.0001, unpaired t-test

Article Snippet: IHC was performed with antibodies (Table ) against RBPMS (RGC soma marker), NEFL (RGC axon marker), GFAP (astrocyte marker) and IBA1 (microglia/macrophage marker) using a LeicaBond RX stainer (Leica Biosystems, Deer Park, IL, USA).

Techniques: Standard Deviation

Journal: Acta Neuropathologica Communications

Article Title: Multiparametric grading of glaucoma severity by histopathology can enable post-mortem substratification of disease state

doi: 10.1186/s40478-024-01880-2

Figure Lengend Snippet: Spearman’s correlation analysis. A Spearman’s correlation demonstrated maximal correlation between RBPMS and NEFL in glaucomatous donors, with minimal correlation seen when comparing other markers in glaucomatous and non-glaucomatous donor tissues. B No statistically significant correlation was observed when comparing GFAP and IBA1 immunoreactivity in the retina for both non-glaucomatous and glaucomatous donors

Article Snippet: IHC was performed with antibodies (Table ) against RBPMS (RGC soma marker), NEFL (RGC axon marker), GFAP (astrocyte marker) and IBA1 (microglia/macrophage marker) using a LeicaBond RX stainer (Leica Biosystems, Deer Park, IL, USA).

Techniques:

Journal: Acta Neuropathologica Communications

Article Title: Multiparametric grading of glaucoma severity by histopathology can enable post-mortem substratification of disease state

doi: 10.1186/s40478-024-01880-2

Figure Lengend Snippet: Post-hoc analysis after stratification of glaucomatous donors. Progressive RGC degeneration and increased glial reactivity were observed in the retina with disease severity. A Loss of RBPMS + RGC cells, B loss of axons (NEFL), C increased astrocytic activation (GFAP), and D microglia/macrophage activation (IBA1) was stratified by disease state. Horizontal bars indicate mean ± standard deviation. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, One-way Analysis of variance (ANOVA) followed by Tukey’s multiple comparison test

Article Snippet: IHC was performed with antibodies (Table ) against RBPMS (RGC soma marker), NEFL (RGC axon marker), GFAP (astrocyte marker) and IBA1 (microglia/macrophage marker) using a LeicaBond RX stainer (Leica Biosystems, Deer Park, IL, USA).

Techniques: Activation Assay, Standard Deviation, Comparison

Journal: Acta Neuropathologica Communications

Article Title: Multiparametric grading of glaucoma severity by histopathology can enable post-mortem substratification of disease state

doi: 10.1186/s40478-024-01880-2

Figure Lengend Snippet: Schematic of donor eye grading workflow. Conceptual outline of steps in acquisition and grading of post-mortem donor eyes, accompanied by qualitative descriptions of protein marker stain changes from non-glaucoma samples in each severity grade. Note that the evaluation of protein markers is confined to the temporal retina. One arrow indicates a mild/minimal change, two arrows indicate a moderate change, and three arrows indicate a severe/maximal change. The predominant/distinguishing features of each grade are: mild – pre-mortem history of glaucoma; moderate – maximal GFAP positivity in temporal neural retina; severe – maximal loss of RBPMS and NEFL positivity in temporal retina

Article Snippet: IHC was performed with antibodies (Table ) against RBPMS (RGC soma marker), NEFL (RGC axon marker), GFAP (astrocyte marker) and IBA1 (microglia/macrophage marker) using a LeicaBond RX stainer (Leica Biosystems, Deer Park, IL, USA).

Techniques: Marker, Staining