Journal: Scientific Reports

Article Title: Munc18-1 is crucial for photoreceptor function, survival and regulation of syntaxin-3 localization and expression

doi: 10.1038/s41598-026-57827-1

Figure Lengend Snippet: Effective removal of Munc18-1 by CRX-cre leads to degeneration between postnatal day 14 and 22. ( A ) Left: Antibody staining for Munc18-1 (green) using rabbit polyclonal antibody against PSD95 (red) for presynaptic terminals in P9 control and P9 Munc18-1 retina. Munc18-1 is present throughout the retina in the outer nuclear layer (ONL), outer plexiform layer (OPL), inner nuclear layer (INL), inner plexiform layer (IPL), and ganglion cell layer (GCL). In control retina, a meshed/plasmalemmal pattern of Munc18-1 is observed surrounding photoreceptor soma. Munc18-1 fluorescence is lost in the photoreceptor and bipolar cells of Munc18-1 cKO mice. Retina region for all represented images is 0.5–1 mm away from optic nerve. All images are maximum intensity projections. Scale bar = 20 μm. Right: Antibody staining for Munc18-1 (green) using mouse monoclonal antibody and PSD95 (red) in P9 control and Munc18-1 cKO retina. A decrease in Munc18-1 fluorescence is signal is observed in Munc18-1 cKO mice. Retina region for all represented images is 0.5–1 mm away from optic nerve. All images are maximum intensity projections. Scale bar = 50 μm. ( B ) Antibody staining for Munc18-1 (green) using rabbit polyclonal antibody against PSD95 (red) for presynaptic terminals in P12 and P14 control and Munc18-1 cKO retina. Top: scale bar = 50 μm. Bottom: scale bar = 20 μm. All images are maximum intensity projections. ( C ) Quantification of outer nuclear layer (ONL) thickness in control, Munc18-1 cKO (green, squares) mice and SNAP-25 cKO (purple, circles) mice. Control mice for both lines are their respective flox/flox littermates, i.e. SNAP-25 flox/flox vs. SNAP-25 cKO and Munc18-1 flox/flox vs. Munc18-1 cKO. SNAP-25 cKO photoreceptors are rapidly lost between P9 and P14 while Munc18-1 cKO photoreceptors are rapidly lost between P14 and P22. Although Munc18-1 flox/flox control ONLs were thicker than Munc18-1 cKO ONLs, they were not statistically significant at any age before P14 (two-way ANOVA; P9, p = 0.9976; P12, p = 0.1992; P14, p = 0.5107). SNAP-25 exhibit significant degeneration after P9 (two-way ANOVA; P9, p = 0.7049; P12, p < 0.0001; P14, p < 0.0001). Error bar represents SEM. ( D ) Averaged scotopic (dark-adapted) and photopic (light-adapted) electroretinogram traces across 7 light intensities in P14 control and P14 Munc18-1 cKO mice (solid line). Arrow indicate stimulation onset, A indicates a-wave, B indicates b-wave. Munc18-1 cKO mice exhibit diminished light responses at all light intensities. Each data point represents one of eye each mouse; one eye was excluded from a control mouse due to eyelid deformity. Error bar represents SEM.

Article Snippet: Mouse polyclonal PKCɑ , Santa Cruz Biotechnology , sc-8393.

Techniques: Staining, Control, Fluorescence

Journal: Scientific Reports

Article Title: Munc18-1 is crucial for photoreceptor function, survival and regulation of syntaxin-3 localization and expression

doi: 10.1038/s41598-026-57827-1

Figure Lengend Snippet: Verification of syntaxin-3 mislocalization by a different syntaxin-3 antibody. ( A ) Munc18-1 (green) and syntaxin-3 (rabbit polyclonal (pc) from ProteinTech) (red) stain in P12 control and Munc18-1 cKO retina. The loss of syntaxin-3 and punctate accumulation in the ONL (indicated by yellow arrowheads) were replicated with this antibody. ( B ) Syntaxin-3 (pc) (red) and RIBEYE (white) stain in P12 control and Munc18- 1 cKO retina. Similar finding on synaptic localization of syntaxin-3 using an independent antibody. All images are maximum intensity projections. ( C ) Punctate syntaxin-3 staining in the ONL. Two-tailed two-sample t-test. ONL: t(5) = − 5.31878; p = 0.00314. ( D ) Syntaxin-3 (pc) signal intensity is decreased in both ONL and the OPL (marked by the presence of RIBEYE). n = 3 in control, n = 4 in cKO. Two-tailed two-sample t-test. ONL: t(5) = 13.13238; p < 0.0001. OPL: t(5) = 3.6032; p = 0.01549.

Article Snippet: Mouse polyclonal PKCɑ , Santa Cruz Biotechnology , sc-8393.

Techniques: Staining, Control, Two Tailed Test

Journal: Scientific Reports

Article Title: Munc18-1 is crucial for photoreceptor function, survival and regulation of syntaxin-3 localization and expression

doi: 10.1038/s41598-026-57827-1

Figure Lengend Snippet: Reagents and resources.

Article Snippet: Mouse polyclonal PKCɑ , Santa Cruz Biotechnology , sc-8393.

Techniques: Western Blot, Recombinant, Tissue, In Situ, Software, Microscopy, Glass