Journal: bioRxiv

Article Title: A paradoxical relationship between mitochondrial calcium regulation and retinal ganglion cell degeneration after axon damage

doi: 10.64898/2026.05.13.724793

Figure Lengend Snippet: (A) Example in vivo 2-photon max intensity projections (mip) of cytoplasmic (Cyto, left) and mitochondrial (Mito, right) Twitch-2b expressed in retinal ganglion cells (RGCs) of VGlut2-Cre mice. (B) Swarm plots of homeostatic cyto- and mito-Ca 2+ levels, as measured by cpVenus (YFP) to mCerulean (CFP) FRET ratios, MWU test (cyto-T2b n = 868 RGCs from 14 retinas and 8 mice, mito-T2b n = 1066 RGCs from 19 retinas and 15 mice). (C) Average intensity projections (aip) of mito-T2b in vivo of the same RGCs at baseline and 4 days later. Magenta arrows indicate RGCs with higher baseline mito-Ca 2+ and green arrows indicate RGCs with lower mito-Ca 2+ levels. (D) Scatterplot of mito-Ca 2+ levels pre and 4 day follow-up timepoints. R = 0.45, (n = 205 RGCs from 5 retinas and 3 mice). (E) Swarm plots of change in mito-Ca 2+ levels from baseline to 4 day follow-up replotted from panel (D). (F) Example aips of mito-T2b at baseline and 10 min following intravitreal Ru265 injection. White arrows indicate RGCs with reduced mito-Ca 2+ . (G) Line graphs of mito-Ca 2+ levels before and 10 min after Ru265 injection. Individual RGCs are in blue, and mean is in black (n = 175 RGCs from 4 retinas and 4 mice). (H) Example aips of cyto-T2b at baseline and 10 min after intravitreal Ru265. White arrows indicate RGCs with increased cyto-Ca 2+ . (I) Line graphs of cyto-Ca 2+ levels before and 10 min after Ru265 injection. Individual RGCs are in orange, and mean is in black (n = 123 RGCs from 4 retinas and 4 mice). (J) Swarm plot comparing changes in Ca 2+ levels after Ru265 between cyto- and mito-T2b. Black bars are mean +/- SEM replotted from panels (G and I). (K) Example aips of KCNG4-Cre mito-T2b acquired with in vivo transscleral imaging. GCL = ganglion cell layer, IPL = inner plexiform layer, NFL = nerve fiber layer. Box indicates magnified region. (L) Representative confocal mips of fixed retinal wholemounts showing endogenous mito-T2b (yellow) and TOMM20 (cyan) immunostaining. Scale bars = 100 μm.

Article Snippet: For overexpression experiments, VGlut2-Cre mice were crossed with B6N.Cg-Gt(ROSA)26Sortm1(CAG-EGFP*)Thm/J (032675, Jackson Labs) mice for outer mitochondrial membrane-GFP expression (not analyzed/used).

Techniques: In Vivo, Injection, Imaging, Immunostaining

Journal: bioRxiv

Article Title: A paradoxical relationship between mitochondrial calcium regulation and retinal ganglion cell degeneration after axon damage

doi: 10.64898/2026.05.13.724793

Figure Lengend Snippet: (A) Example in vivo aips of mito-T2b from VGlut2-Cre transgenic mouse and the same imaged region in fixed retinal wholemount immunostained for SPP1 (red, αRGCs) and TBR2 (cyan, ip-RGCs). Mito-T2b is greyscale. Arrows represent cells identified as positive for the different stains: cyan = TBR2 only, red = SPP1 only, and white = SPP1 / TBR2 positive. (B) Swarm plots of mito-Ca 2+ levels in individual RGCs positive for indicated RGC markers. Means are labeled with a black bar +/- SEM (n = 479 RGCs from 6 retinas and 6 mice). (C) Representative aips of mito-T2b expression in VGlut2-Cre (all RGCs) and KCNG4-Cre transgenic mice (αRGCs). (D) Swarm plots of mito-Ca 2+ levels from VGlut2-Cre and KCNG4-Cre transgenic mice (n = 79 RGCs from 6 retinas and 3 mice). Scale bars = 100 μm.

Article Snippet: For overexpression experiments, VGlut2-Cre mice were crossed with B6N.Cg-Gt(ROSA)26Sortm1(CAG-EGFP*)Thm/J (032675, Jackson Labs) mice for outer mitochondrial membrane-GFP expression (not analyzed/used).

Techniques: In Vivo, Transgenic Assay, Labeling, Expressing

Journal: Frontiers in Behavioral Neuroscience

Article Title: iMOSS: an integrated open-source tail suspension test platform for high-resolution immobility scoring and synchronization with neural activity

doi: 10.3389/fnbeh.2026.1819512

Figure Lengend Snippet: Behavioral scoring synchronization with neural activity in a single mouse. (A) Right: Representative image of a VGluT2-Cre mouse during the TST with patch cable coupled to the optical fiber cannula implanted in the medial septum (MS). Left: Green dots show GCaMP8s expression of MS VGluT2 neurons, while blue shows nuclear DAPI staining. (B) Load-cell signals detected by iMOSS-AS (top), manually scored mobility (red) and immobility (blue) bouts with iMOSS-MV (middle), and GCaMP8s signals recorded from MS vGluT2 neurons with calcium peaks (red crosses) (bottom). (C) Mean ( ± SEM) calcium Z score (left), peak frequency (middle) and amplitude (right) between immobility and mobility bouts. (D) Heatmap (top) and mean trace (bottom) of GCaMP8s signals within 1-s before and after the onset of mobility and immobility, scored by iMOSS-AS. The white regions inside the heatmap indicate masked time points where the aligned window exceeded the target event boundaries. Only event-pure segments were included in the analysis to avoid contamination from adjacent behavioral states, and averages were computed from valid samples only. (E) Raster plots (top) and line graphs (bottom) illustrating the frequency and amplitude of calcium peaks relative to mobility and immobility onsets. (F) Mean ( ± SEM) calcium peak count (left) and amplitude (right) within 1-s before and after the onset of immobility and mobility bouts. * p < 0.05, ** p < 0.01.

Article Snippet: The C57BL/6 mice were purchased from Jackson Laboratory (Bar Harbor, ME), while transgenic vGluT2-Cre mice were obtained from Jackson Lab then bred at the National Institute on Drug Abuse Intramural Research Program animal facility.

Techniques: Activity Assay, Expressing, Staining