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: Developmental biology

Article Title: Wild-type cells rescue genotypically Math1-null hair cells in the inner ears of chimeric mice.

doi: 10.1016/j.ydbio.2007.02.028

Figure Lengend Snippet: Fig. 3. Examples of P4.5 Math1-null chimeric and control mice. Panels A–D show HCs in P4.5 mice where β-Gal histochemistry was used to mark the genotypically Math1-null or Math1+/β-Gal cells and myosin VIIa immunocy- tochemistry was used to identify HCs. Double-labeled mutant HCs are shown in the cochlea (arrows in panel A), and in the crista (arrows in panel B) of Math1+/β-Gal chimeras. Similarly double-labeled HCs are also found in the cochlea (C) and crista (D) of Math1-null chimeric mice. Panels E–H show examples of DNA in situ hybridization where labeled cells possess an obvious dark blue/purple dot within the nucleus while the nuclei appear green due to counterstaining with methyl green. The GT probe marks the wild-type HCs (arrowheads in panels E–H), while the mutant HCs remain unlabeled (arrows in panels E–H). All HCs are labeled with DNA in situ hybridization in GT control mice (E), while no HCs are labeled in wild-type mice (F). Examples of genotypically mutant cells are shown in the macula (G) and crista (H) of Math1- null chimeric mice at P4.5. Scale bar=50 μm (in panel D for panels A–D, in panel H for panels E–H).

Article Snippet: The sections were incubated with goat anti-myosin VIIa IgG (Santa Cruz Biotechnology, Inc., Santa Cruz, CA) at a dilution of 1:20 and rabbit anti-β-Gal IgG (Cortex Biochem, San Leandro, CA) at a dilution of 1:25.

Techniques: Control, Labeling, Mutagenesis, DNA In Situ Hybridization

Journal: Developmental biology

Article Title: Wild-type cells rescue genotypically Math1-null hair cells in the inner ears of chimeric mice.

doi: 10.1016/j.ydbio.2007.02.028

Figure Lengend Snippet: Fig. 4. Examples of Myosin VIIa and β-Gal immunolabeling in HCs in Math1-null chimeric and control mice at P2.5. Images in column 3 (anti-myosin VIIa/anti-β- Gal) are the merged images from column 1 (anti-myosin VIIa) and column 2 (anti-β-Gal). Myosin VIIa is used to label HCs while β-Gal is used to detect cells that are missing at least one copy of the Math1 gene. HCs are labeled only with the anti-myosin VIIa antibody in the cochlea (C) and crista (F) of wild-type mice. Double labeled HCs are shown in the cochlea (arrow in panel I) and crista (arrow in panel L) of Math1+/β-Gal chimeric mice to demonstrate the pattern of intermingling observed in chimeric mice. In Math1-null chimeras, double-labeled mutant cells (arrows in panels O and R) are found intermixed with wild-type cells which are labeled only with anti-myosin VIIa. Double-labeled mutant HCs are shown in the cochlea (arrows in panel O) and crista (arrow in panel R) in the mutant chimeras. The tangential plane of section in panels M–O results in two layers of HCs being visible and the lack of a full view of the HC cytoplasm. Green fluorescence has been found on top of most wild-type and mutant HCs (arrowheads in panels B, E, H, K, N and Q), suggesting an autofluorescence from sterocillia and cuticular plates. Scale bar=50 μm (in panel O for panels A–C, G–I and panels M–O). Scale bar=15 μm (in panel R for panels D–F, J–L and panels P–R).

Article Snippet: The sections were incubated with goat anti-myosin VIIa IgG (Santa Cruz Biotechnology, Inc., Santa Cruz, CA) at a dilution of 1:20 and rabbit anti-β-Gal IgG (Cortex Biochem, San Leandro, CA) at a dilution of 1:25.

Techniques: Immunolabeling, Control, Labeling, Mutagenesis, Fluorescence