Journal: iScience

Article Title: Endolysosomal dysfunction impairs proteostasis and induces neurodegeneration in vivo

doi: 10.1016/j.isci.2025.113460

Figure Lengend Snippet: GFP-Rab5 Q79L mice exhibit motor deficits (A) Hindlimb clasping scores for GFP control mice (squares) and GFP-Rab5 Q79L mice (circles) at the indicated ages. (B) Number of falls in 2 min during the hanging wire behavioral assay for 3-month-old GFP and GFP-Rab5 Q79L mice. (C) Western blots of mouse brain lysates confirm expression of 3-month-old GFP and GFP-Rab5 Q79L mice. Upper panel: immunoblot using an anti-Rab5 antibody; lower panel: immunoblot using an anti-GFP antibody. Open arrow: GFP-Rab5 Q79L ; solid arrow: Rab5 or GFP. Cropped blots are shown in full in . (D) Quantification of Rab5 protein expression levels from (C). n = 9 and 10 mice in the GFP group and in the GFP-Rab5 Q79L group, respectively. All behavioral tests were repeated three times per mouse. Data are shown as the mean ± SEM. ∗∗∗ p < 0.001 and ∗∗∗∗ p < 0.0001; two-way ANOVA (A) and unpaired two-tailed t test (B and D).

Article Snippet: Accordingly, GFP-Rab5 Q79L mice will prove useful in expanding our understanding of endolysosomal dysfunction in proteostasis and pTDP-43 pathology.

Techniques: Control, Behavioral Assay, Western Blot, Expressing, Two Tailed Test

Journal: iScience

Article Title: Endolysosomal dysfunction impairs proteostasis and induces neurodegeneration in vivo

doi: 10.1016/j.isci.2025.113460

Figure Lengend Snippet: Rab5 Q79L induces endosomal enlargement and abnormal lysosome morphology, recapitulating endolysosomal dysfunction (A) Immunofluorescence for GFP (green) and early endosome antigen 1 (EEA1; red), an early endosome marker, in the cortex of 3-month-old GFP and GFP-Rab5 Q79L mice. DAPI marks nuclei in blue. Scale bar is 20 μm. (B) Approximate size of endosomes calculated by measuring the area of EEA1-positive vacuoles from the immunofluorescence images in A n = 5 mice per group. (C) Immunohistochemistry for the lysosome marker cathepsin D (CatD) in the cortex of 3-month-old GFP and GFP-Rab5 Q79L mice. Arrows: abnormal CatD puncta. Slides were stained with hematoxylin. Scale bar is 60 μm. (D) The percentage of cells with enlarged CatD-positive puncta is quantified; n = 5 mice per group. (E and F) (E) Immunofluorescence for GFP (green) and CatD (red) in the cortex of 3-month-old GFP and GFP-Rab5 Q79L mice with corresponding quantification (F). DAPI marks nuclei in blue. Scale bar is 10 μm. (G) Electron microscopy of the brains of 3-month-old GFP and GFP-Rab5 Q79L mice. Scale bars on left, middle, and right columns are 5, 1, and 500 nm, respectively. Black arrows indicate enlarged endolysosomes. Data are shown as the mean ± SEM. ∗∗∗∗ p < 0.0001, unpaired two-tailed t test.

Article Snippet: Accordingly, GFP-Rab5 Q79L mice will prove useful in expanding our understanding of endolysosomal dysfunction in proteostasis and pTDP-43 pathology.

Techniques: Immunofluorescence, Marker, Immunohistochemistry, Staining, Electron Microscopy, Two Tailed Test

Journal: iScience

Article Title: Endolysosomal dysfunction impairs proteostasis and induces neurodegeneration in vivo

doi: 10.1016/j.isci.2025.113460

Figure Lengend Snippet: GFP-Rab5 Q79L mice exhibit defects in proteostasis (A) Immunohistochemistry for ubiquitin in the cortex of 3-month-old GFP and GFP-Rab5 Q79L mice. Slides were stained with hematoxylin. Scale bar is 60 μm. (B) The percentage of cells with ubiquitin aggregates is quantified; n = 5 mice per group. (C and D) (C) Immunofluorescence for GFP (green) and ubiquitin (red) in the cortex of 3-month-old GFP and GFP-Rab5 Q79L mice with corresponding quantification (D). DAPI marks nuclei in blue. Scale bar is 10 μm. (E) Immunohistochemistry for p62 in the cortex of 3-month-old GFP and GFP-Rab5 Q79L mice. Slides were stained with hematoxylin. Scale bar is 60 μm. (F) The percentage of cells with p62 aggregates is quantified; n = 5 per group. (G and H) (G) Immunofluorescence for GFP (green) and p62 (red) in the cortex of 3-month-old GFP and GFP-Rab5 Q79L mice with corresponding quantification (H). DAPI marks nuclei in blue. Scale bar is 10 μm. Data are shown as the mean ± SEM. ∗∗∗∗ p < 0.0001, unpaired two-tailed t test.

Article Snippet: Accordingly, GFP-Rab5 Q79L mice will prove useful in expanding our understanding of endolysosomal dysfunction in proteostasis and pTDP-43 pathology.

Techniques: Immunohistochemistry, Ubiquitin Proteomics, Staining, Immunofluorescence, Two Tailed Test

Journal: iScience

Article Title: Endolysosomal dysfunction impairs proteostasis and induces neurodegeneration in vivo

doi: 10.1016/j.isci.2025.113460

Figure Lengend Snippet: Rab5 Q79L induces neurodegeneration and neuroinflammation in vivo (A–F) Representative images and quantification of immunohistochemistry for the neuronal (NeuN, A and B), astrocytic (GFAP, C and D), and microglial (Iba1, E and F) markers in the cortex of 3-month-old GFP and GFP-Rab5 Q79L mice. Slides were stained with hematoxylin. Scale bars in (A) are 300 μm (NeuN). Scale bars in (C) and (E) are 60 μm (GFAP and Iba1). n = 6 mice per group in panels (B), (D), and (F). Data are shown as the mean ± SEM. ∗∗∗ p < 0.001 and ∗∗∗∗ p < 0.0001, unpaired two-tailed t test.

Article Snippet: Accordingly, GFP-Rab5 Q79L mice will prove useful in expanding our understanding of endolysosomal dysfunction in proteostasis and pTDP-43 pathology.

Techniques: In Vivo, Immunohistochemistry, Staining, Two Tailed Test

Journal: iScience

Article Title: Endolysosomal dysfunction impairs proteostasis and induces neurodegeneration in vivo

doi: 10.1016/j.isci.2025.113460

Figure Lengend Snippet: Rab5 Q79L induces TDP-43 proteinopathy but not TDP-43 mislocalization nor loss of cryptic splicing in vivo (A and B) (A) Immunohistochemistry for TDP-43 phosphorylated at serines 409 and 410 (pTDP-43) in the cortex and thalamus of 3-month-old GFP-Rab5 Q79L mice, with ordinal score (B). Slides were stained with hematoxylin. Scale bars are 60 μm. (C) Immunofluorescence for GFP (green) and pTDP-43 (red) in the cortex of 3-month-old GFP and GFP-Rab5 Q79L mice. DAPI marks nuclei in blue. Scale bar is 10 μm. (D) Immunohistochemistry for total TDP-43 in the cortex and thalamus of 3-month-old GFP-Rab5 Q79L mice. (E) Cryptic exon inclusion within TDP-43 target transcripts Sortilin1 and Ap3b2 between 3-month-old GFP and GFP-Rab5 Q79L mice. n = 6 mice per group. Data are shown as the mean ± SEM. ∗∗∗ p < 0.001 and ∗∗∗∗ p < 0.0001; ns, not significant; unpaired two-tailed t test.

Article Snippet: Accordingly, GFP-Rab5 Q79L mice will prove useful in expanding our understanding of endolysosomal dysfunction in proteostasis and pTDP-43 pathology.

Techniques: In Vivo, Immunohistochemistry, Staining, Immunofluorescence, Two Tailed Test

Journal: iScience

Article Title: Endolysosomal dysfunction impairs proteostasis and induces neurodegeneration in vivo

doi: 10.1016/j.isci.2025.113460

Figure Lengend Snippet: Rab5 Q79L induces morphological alterations in the nuclear envelope and nuclear pore complex (A) Immunofluorescence for GFP (green) and Lamin B1 (red) in the cortex of 3-month-old GFP and GFP-Rab5 Q79L mice. DAPI marks nuclei in blue. Scale bar is 10 μm. (B) The percentage of cells with abnormal Lamin B1 staining. n = 3 mice per group. (C) Immunofluorescence for GFP (green) and nuclear pore complex (NPC, red) in the cortex of 3-month-old GFP and GFP-Rab5 Q79L mice. DAPI marks nuclei in blue. Scale bar is 10 μm. (D) The percentage of cells with abnormal NPC staining. n = 3 mice per group. Data are shown as the mean ± SEM. ∗∗∗∗ p < 0.0001 and ∗∗ p < 0.01; unpaired two-tailed t test.

Article Snippet: Accordingly, GFP-Rab5 Q79L mice will prove useful in expanding our understanding of endolysosomal dysfunction in proteostasis and pTDP-43 pathology.

Techniques: Immunofluorescence, Staining, Two Tailed Test