Journal: bioRxiv

Article Title: The App NL-G-F mouse retina is a site for preclinical Alzheimer’s disease diagnosis and research

doi: 10.1101/2020.07.25.220707

Figure Lengend Snippet: (a) Representative electroretinogram measurements, with increasing light intensities, in 18-month-old App NL-G-F (green) and WT (blue) mice. (b) The amplitude of the a-wave is unaffected in App NL-G-F mice of 18 months, yet (c) the latency time is decreased for the lowest light intensity. Two-way ANOVA with Sidak’s multiple comparisons test (F 1,85 = 23.14 for the effect of genotype, p<0.0001); n=8-10. (d) Oscillatory potentials show a similar wave front (measured as the area under the curve, AUC), but (e) a reduced latency in 18-month-old App NL-G-F mice, compared to WT. Two-way ANOVA with Sidak’s multiple comparisons test (F 1,80 = 37.14 for the effect of genotype, p=0.0013, 0.0124, 0.0378 for 0.01, 0.1 and 1 cd*s/m 2 respectively); n=8-10. (f) Similarly, the amplitude of the b-wave is unaltered, but (g) the latency time is decreased in 18-month-old App NL-G-F mice compared to WT mice. Two-way ANOVA with Sidak’s multiple comparisons test (F 1,97 = 27.90 for the effect of genotype, p=0.0206 and 0.044 for 0.1 and 1 cd*s/m 2 ); n=8-10. Full electroretinogram data is given in Supplementary figure 2. (h) Analysis of the thickness of the retinal layers, as measured by optical coherence tomography, shows no differences between App NL-G-F and WT mice from 2 to 18 months of age, (i) with the exception of a thinning of the outer nuclear layer. Two-way ANOVA with Sidak’s multiple comparisons test (F 1,106 = 32.14 for the effect of genotype, p=0.0016 and p<0.0001 for 12 and 18 months, respectively); n=8-9. (j-k) Representative optical coherence tomography images of WT (j) and App NL-G-F (k) retinas. Dotted lines delineate the outer nuclear layer. (l) Quantification of RBPMS + retinal ganglion cell density reveals ganglion cell loss in 18-month-old App NL-G-F compared to WT mice. Unpaired two-tailed t-test (t 14 =2.745,p=0.0158); n=8. (m-n) Cell counts of melanopsin + ganglion cells (m) and ChAT + neurons (n) show no differences in numbers between 18-month-old App NL-G-F and WT mice. (o) A longitudinal study of visual acuity in App NL-G-F mice from 3 till 18 months revealed a decrease with age but no genotype differences. Two-way ANOVA (F 10,180 =21.11 for the effect of age, F 1,180 =0.1149 for the effect of genotype); n=9. Scale bar: 25 µm. Data are depicted as mean ± SEM; RNFL = retinal nerve fiber layer, GCL = ganglion cell layer, IPL = inner plexiform layer, INL = inner nuclear layer, OPL = outer plexiform layer, ONL = outer nuclear layer, PL = photoreceptor layer.

Article Snippet: For wholemount (immuno)staining, wholemounts were frozen for 15 min at −80°C, and incubated overnight with primary antibodies for melanopsin (1:5000, #AB-N38, Advanced Targeting Systems), ionized calcium-binding adapter molecule 1 (Iba1; 1:1000, #019-19741, Wako), choline acetyltransferase (ChAT; 1:500, #AB144P, Millipore), platelet-derived growth factor receptor beta (PDGFRβ) (1:500, #ab32570, Abcam), GFAP (1:1000, #Z0334, DAKO), RNA-binding protein with multiple splicing (RBPMS) (1:250, #1830-RBPMS, PhosphoSolutions), protein kinase C alpha (PKCα) (1:100, #sc-10800, Santa Cruz), green fluorescent protein (GFP; 1:500, #ab13970, Abcam) and/or Aβ (6E10, 1:500, #803001, BioLegend and 82E1, 1:300, #10323, Immuno-Biological Laboratories), or with biotinylated isolectin B4 (1:50, #B-1205, Vector Laboratories), followed by incubation with corresponding Alexa-conjugated secondary antibodies or Alexa-conjugated streptavidin (Invitrogen).

Techniques: Tomography, Two Tailed Test

Journal: Cell reports

Article Title: A non-canonical retina-ipRGCs-SCN-PVT visual pathway for mediating contagious itch behavior

doi: 10.1016/j.celrep.2022.111444

Figure Lengend Snippet:

Article Snippet: The primary antibodies used were rabbit anti-melanopsin (1:2000, Advanced Targeting Systems, AB-N38), rabbit anti-GRP (1:500, Immunostar, 20073), chicken anti-GFP (1:400, Aves Labs, GFP-1020), and mouse anti-mCherry (1:1000, Developmental Studies Hybridoma Bank, 3A11).

Techniques: Virus, Plasmid Preparation, Recombinant, Enzyme-linked Immunosorbent Assay, RNAscope, Multiplex Assay, Software