Journal: Scientific Reports

Article Title: Fatty acid binding protein type 7 deficiency preserves auditory function in noise-exposed mice

doi: 10.1038/s41598-023-48702-4

Figure Lengend Snippet: Experimental design, body weights, and FABP7 expression in the cochlea. Schematic of the experiments using wild-type (WT) and Fabp7 knockout (KO) mice ( A ). The body weight (BW) of Fabp7 KO mice was not significantly different from that of WT mice ( B ). Immunohistochemistry of FABP7 in the cochlea of WT mice revealed that FABP7 was expressed throughout the spiral ganglion (SG), organ of Corti (OC), spiral limbus (SLim), and spiral ligament (SLig) ( C ). Scale bar, 200 μm ( C – D ). No expression of FABP7 was observed in the cochlea of Fabp7 KO mice ( D ). Maximum projection images of the OC showing high expression of FABP7 in Schwann cells and SOX2-positive supporting cells around the MYO7a-positive outer hair cells (OHCs) in the cochlea of WT mice ( E ). In the SG, FABP7 was expressed in satellite cells surrounding TUJ-1-positive neurons ( F ). Scale bars, 50 μm ( E – F ). Auditory brainstem response; ABR, weeks; W, post-noise exposure day; PNED, inner hair cell; IHC. Statistical significance was determined using two-way analysis of variance, followed by Šídák multiple comparison test. Error bars represent standard deviation.

Article Snippet: After rinsing with PBS, tissue sections were blocked with 3% bovine serum albumin/0.3% Triton X-100/ PBS for 30 min at room temperature, the unconjugated AffiniPure Fab fragment of anti-mouse IgG (1:10, Jackson ImmunoResearch, West Grove, PA) for 2 h at room temperature, and incubated overnight with the following primary antibodies: (1) rabbit IgG anti-FABP7 (1:1000, Merck, Darmstadt, Germany #PRS4259), (2) mouse IgG anti-TUJ-1 (1:500, Abcam, Cambridge, UK #ab78078), (3) mouse IgG2a anti-Myosin 7a (MYO7a) (1:200, Santa Cruz Biotechnology, Dallas, TX, USA #SC-74516), and (4) goat IgG anti-SOX2 (1:500, R&D Systems, Minneapolis, MN, USA #AF2018).

Techniques: Expressing, Knock-Out, Immunohistochemistry, Comparison, Standard Deviation

Journal: Scientific Reports

Article Title: Gata3 is required in late proneurosensory development for proper sensory cell formation and organization

doi: 10.1038/s41598-023-39707-0

Figure Lengend Snippet: Deletion of Gata3 results in loss of HCs in a basal to apical gradient ( A – F′ ) Representative images from the basal, middle, and apical regions of the cochlea for HCs indicated by MYOSIN7A + staining. Two different controls were used, Gata3 f/f and Sox2-cre ERT2 , in order to account for the haploinsufficent phenotype of the Cre line used. Both the heterozygous and homozygous mutant show IHC duplets (white circles) and missing rows of OHCs (white brackets), while the homozygous mutant also shows ectopic HCs in the GER. ( G ) Total hair cell quantification was performed in the base, middle, and apex in 100 µm sections using a One-way ANOVA with post hoc Dunnett’s test (P** ≤ 0.01; P*** ≤ 0.001) Scale bar: 50 µm.

Article Snippet: The following primary antibodies were used: MYO6 Rabbit (Sigma; 1:1000), MYOSIN7A Mouse (DSHB; 1:200), MYSOIN7A Rabbit (Proteus Biosciences, Inc.; 1:500), Neurofilament 200 (NF200) HC Chicken (Aves; 1:200), and SOX2 Rabbit (Sigma; 1:500).

Techniques: Staining, Mutagenesis

Journal: The Journal of Neuroscience

Article Title: The Severity of Infection Determines the Localization of Damage and Extent of Sensorineural Hearing Loss in Experimental Pneumococcal Meningitis

doi: 10.1523/JNEUROSCI.0554-16.2016

Figure Lengend Snippet: Confocal microscopy analysis of IHC presynaptic ribbons 3 weeks after infection. A, B, Representative confocal images (maximal projection from a focal series) of IHC synapses from the basal turn of a control (A) and an infected (high INO; B) cochlea 3 weeks after infection, immunolabeled for presynaptic ribbons (CtBP2, red) and nuclei (DAPI, blue). The dashed lines show the approximate outlines of two IHCs, determined by immunolabeling for hair cells (Myosin7a, yellow; data not shown). C, Quantification of presynaptic ribbons per IHC for the three cochlear regions in control and infected animals (controls, empty squares; low INO, empty circles; intermediate INO, gray circles; high INO, black circles; n = 3 per group). The means are indicated in the bar graphs. An unpaired t test with Welch's correction was used for single comparisons. D, Spearman's correlation between the number of ribbons per IHC and the initial inoculum load in the basal turn. Scale bars: 10 μm. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.

Article Snippet: The hair cell-specific antibody against Myosin7a (polyclonal rabbit anti Myosin-7a, Proteus) together with the presynaptic ribbon-specific antibody against CtBP2 (monoclonal mouse anti CtBP2, BD Bioscience) were both diluted 1:200 in blocking solution and incubated overnight at 4°C.

Techniques: Confocal Microscopy, Infection, Immunolabeling