Journal: Frontiers in Aging Neuroscience

Article Title: Synaptopathy as a Mechanism for Age-Related Vestibular Dysfunction in Mice

doi: 10.3389/fnagi.2019.00156

Figure Lengend Snippet: Loss of calyces in 24-month-old utricular extrastriola. (A–E) Extrastriolar and (F–J) striolar regions of 2, 18 and 24-month-old mouse utricles were analyzed separately after immunostaining with tenascin-C (Tnc) and secreted phosphoprotein 1 (Spp1). (A) Representative confocal images of extrastriolar calyces and type I hair cells. (B) Density of calyx (Tnc+) in 2 ( n = 14), 18 ( n = 9) and 24 ( n = 8) month old utricular extrastriola. (C) Density of type I hair cell associated with calyx in 2 ( n = 9), 18 ( n = 4) and 24 ( n = 4) month old utricular extrastriola. (D) Density of orphan type I hair cell and (E) density of orphan calyx in 2 ( n = 9), 18 ( n = 4) and 24 ( n = 4) month old utricular extrastriola. * p < 0.05 by one-way ANOVA comparing to 2-month-old group followed by Dunn’s post-test. (F) Representative confocal images of striolar calyces and type I hair cells. (G) Density of calyx (Tnc+) in 2 ( n = 15), 18 ( n = 9) and 24 ( n = 8) month old utricular striola. (H) Density of type I hair cell associated with calyx in 2 ( n = 9), 18 ( n = 4) and 24 ( n = 4) month old utricular striola. (I) Density of orphan type I hair cells and (J) density of orphan calyx in 2 ( n = 9), 18 ( n = 4) and 24 ( n = 4) month old utricular striola.

Article Snippet: The micro-dissected utricles were blocked in 5% normal horse serum with 1% Triton X-100 in PBS for 1 h, followed by incubation in primary antibodies (diluted in blocking buffer) at 25°C for 16 h. The primary antibodies used in this study were anti-myosin VIIa (rabbit anti-MyoVIIa, 25-6790, Proteus Biosciences, 1:500), anti-C-terminal binding protein 2 (mouse anti-CtBP2 IgG1, 612044, BD Biosciences, 1:200), anti-glutamate receptor 2 (mouse anti-GluA2 IgG2a, MAB397, Millipore, 1:2,000), anti-tenascin C (rabbit anti-tenascin, AB19013, Millipore, 1:500), anti-secreted phosphoprotein 1 (or osteopontin; goat anti-Spp1, AF808, RnD Systems, 1:500), anti-βIII-tubulin (mouse Tuj1 IgG2a, MMS-435P, BioLegend, 1:1,000) and anti-sox2 (goat anti-sox2, sc-17320, Santa Cruz Biotechnology).

Techniques: Immunostaining

Journal: Frontiers in Aging Neuroscience

Article Title: Synaptopathy as a Mechanism for Age-Related Vestibular Dysfunction in Mice

doi: 10.3389/fnagi.2019.00156

Figure Lengend Snippet: Loss of calyceal synapses but not bouton synapses in 24-month-old utricular extrastriola. (A–E) Extrastriolar and (F–J) striolar regions of 2, 18 and 24-month-old mouse utricles were analyzed separately after immunostaining with tenascin-C (Tnc) and Ctbp2. (A) Representative confocal images of extrastriolar calyces and synaptic ribbons. Density of total synaptic ribbon (B) , noncalyceal ribbon (C) and calyceal ribbon (D) in 2 ( n = 13), 18 ( n = 8) and 24 ( n = 7) month old utricular extrastriola. (E) Number of synaptic ribbons associated with each calyx in 2 ( n = 13), 18 ( n = 8) and 24 ( n = 7) month old utricular extrastriola. * p < 0.05 and ** p < 0.01 by one-way ANOVA comparing to 2-month-old group followed by Dunn’s post-test. (F) Representative confocal images of striolar calyces and synaptic ribbons. Density of total synaptic ribbon (G) , noncalyceal ribbon (H) and calyceal ribbon in (I) 2 ( n = 14), 18 ( n = 7) and 24 ( n = 9) month old utricular striola. (J) Number of synaptic ribbons associated with each calyx in 2 ( n = 14), 18 ( n = 7) and 24 ( n = 9) month old utricular striola.

Article Snippet: The micro-dissected utricles were blocked in 5% normal horse serum with 1% Triton X-100 in PBS for 1 h, followed by incubation in primary antibodies (diluted in blocking buffer) at 25°C for 16 h. The primary antibodies used in this study were anti-myosin VIIa (rabbit anti-MyoVIIa, 25-6790, Proteus Biosciences, 1:500), anti-C-terminal binding protein 2 (mouse anti-CtBP2 IgG1, 612044, BD Biosciences, 1:200), anti-glutamate receptor 2 (mouse anti-GluA2 IgG2a, MAB397, Millipore, 1:2,000), anti-tenascin C (rabbit anti-tenascin, AB19013, Millipore, 1:500), anti-secreted phosphoprotein 1 (or osteopontin; goat anti-Spp1, AF808, RnD Systems, 1:500), anti-βIII-tubulin (mouse Tuj1 IgG2a, MMS-435P, BioLegend, 1:1,000) and anti-sox2 (goat anti-sox2, sc-17320, Santa Cruz Biotechnology).

Techniques: Immunostaining