prolong diamond antifade mountant with 4',6-diamidino-2-phenylindole (dapi)  (Thermo Fisher)

Journal: Neurochemical Research

Article Title: Histamine H3 Receptor Antagonists Influence the Directional Growth of Type II Spiral Ganglion Neurites Within the Developing Cochlea of C57BL/6 Mice

doi: 10.1007/s11064-025-04521-9

Figure Lengend Snippet: Expression and immunolocalization of histamine H3 receptor (H3R) in the cochlea of P4-5 C57BL/6 mice. a Relative H3R mRNA expression in the organ of Corti (OC), lateral wall (LW), and modiolus (MOD). Each sample consisted of tissue from four mice, with three biological replicates per group. b Schematic illustration of a mid-modiolar section of the mouse cochlea highlighting major anatomical structures within the scala media. Reproduced with permission obtained from Springer Nature under CC BY 4.0 . c Low-magnification image of a longitudinal cochlear cryosection. d – e High-magnification views of the boxed regions in c. f Immunofluorescence staining showing co-localization of H3R and phalloidin in both inner and outer HCs within the organ of Corti. H3R (green in c-e, red in f), Phalloidin (green in f), nuclei (DAPI, blue). Scale bars: 200 μm in c , 100 μm in d-e , 10 μm in f

Article Snippet: Nuclear staining was performed using ProLong Gold Antifade Mountant with DAPI (cat. #8961S, Cell Signaling Technology, USA).

Techniques: Expressing, Immunofluorescence, Staining

Journal: Neurochemical Research

Article Title: Histamine H3 Receptor Antagonists Influence the Directional Growth of Type II Spiral Ganglion Neurites Within the Developing Cochlea of C57BL/6 Mice

doi: 10.1007/s11064-025-04521-9

Figure Lengend Snippet: Localization of H3R in hair cells (HCs) and spiral ganglion neurons (SGNs), and its absence at ribbon synapses. a Immunofluorescence staining showing H3R expression in both inner and outer HCs within the organ of Corti. b Negative control incubated only with the secondary antibody (Alexa Fluor™ 488) without the primary anti-H3R antibody. c High-magnification image illustrating H3R signals at the neurite terminals of SGNs near the HCs. d – e Representative images demonstrating H3R expression in SGNs and the corresponding negative control. f High-magnification image showing H3R localization in both the somata and neurites of SGNs. g – i No colocalization is observed between Ribeye and H3R, indicating the absence of H3R at ribbon synapses. H3R (green); neurofilament 200 (NF200, red in a–f); Ribeye (red in g–i); nuclei (DAPI, blue). Scale bars: 15 μm in a – c ; 20 μm in d , e , g ; 5 μm in f ; 10 μm in h , i

Article Snippet: Nuclear staining was performed using ProLong Gold Antifade Mountant with DAPI (cat. #8961S, Cell Signaling Technology, USA).

Techniques: Immunofluorescence, Staining, Expressing, Negative Control, Incubation

Journal: Neurochemical Research

Article Title: Histamine H3 Receptor Antagonists Influence the Directional Growth of Type II Spiral Ganglion Neurites Within the Developing Cochlea of C57BL/6 Mice

doi: 10.1007/s11064-025-04521-9

Figure Lengend Snippet: Effects of H3R antagonist ciproxifan on cochlear hair cell (HC) morphology. a Schematic illustration of cochlear explant treatment with the H3R antagonists. b Representative image of the control group (0µM ciproxifan). c – e Cochlear explants were treated with ciproxifan at concentrations of 10µM, 50µM, and 100µM, respectively. Representative images shown are from the middle turn of the cochlea. HCs are labeled with phalloidin (red), and nuclei with DAPI (blue). Scale bars: 25 μm

Article Snippet: Nuclear staining was performed using ProLong Gold Antifade Mountant with DAPI (cat. #8961S, Cell Signaling Technology, USA).

Techniques: Control, Labeling

Journal: Neurochemical Research

Article Title: Histamine H3 Receptor Antagonists Influence the Directional Growth of Type II Spiral Ganglion Neurites Within the Developing Cochlea of C57BL/6 Mice

doi: 10.1007/s11064-025-04521-9

Figure Lengend Snippet: Effects of H3R antagonist pitolisant on cochlear HCs. a Representative image of the control group (0µM pitolisant). b – d Cochlear explants treated with pitolisant at 10µM, 50µM, and 100µM, respectively. a 4 , b 4 , c 4 , d 4 High-magnification views of the boxed regions in A 3 -D 3 , respectively. e – f Quantification of IHCs and OHCs in the apical, middle, and basal turns showed no statistically significant differences between pitolisant-treated groups and controls ( n = 6 per group). Phalloidin (red), nuclei (DAPI, blue). Scale bars: 25 μm

Article Snippet: Nuclear staining was performed using ProLong Gold Antifade Mountant with DAPI (cat. #8961S, Cell Signaling Technology, USA).

Techniques: Control

Journal: Neurochemical Research

Article Title: Histamine H3 Receptor Antagonists Influence the Directional Growth of Type II Spiral Ganglion Neurites Within the Developing Cochlea of C57BL/6 Mice

doi: 10.1007/s11064-025-04521-9

Figure Lengend Snippet: Dimethyl sulfoxide (DMSO) at experimental concentrations does not affect HC viability or morphology. a Representative image of the control group cultured in culture medium without DMSO, showing intact HC morphology. b – c Explants treated with 0.50% and 1.00% DMSO, respectively, exhibited no detectable HC loss or structural alterations. d – e Quantification of IHCs and OHCs in the apical, middle, and basal turns showed no statistically significant differences between DMSO-treated and control groups. Phalloidin (red), nuclei (DAPI, blue). ns > 0.05; n = 6 per group. Scale bar: 50 μm

Article Snippet: Nuclear staining was performed using ProLong Gold Antifade Mountant with DAPI (cat. #8961S, Cell Signaling Technology, USA).

Techniques: Control, Cell Culture

Journal: Neurochemical Research

Article Title: Histamine H3 Receptor Antagonists Influence the Directional Growth of Type II Spiral Ganglion Neurites Within the Developing Cochlea of C57BL/6 Mice

doi: 10.1007/s11064-025-04521-9

Figure Lengend Snippet: H3R antagonists induce misdirected neurite outgrowth in type II SGNs in cochlear explants. a Representative immunofluorescence images showing typical SGN innervation: type I SGNs project radially to IHCs, while type II SGNs extend longitudinally from the apex toward the base to innervate OHCs. b Cochlear explant treated with 50µM ciproxifan showing misdirected type II SGN neurites. c Cochlear explant treated with 50µM pitolisant showing a similar reversal of type II SGN projections. a 4 , b 4 , c 4 High-magnification views of the boxed regions in a 3 , b 3 , c 3 . White asterisks indicate misdirected neurites projecting toward the cochlear apex. d Percentage of misdirected type II SGNs following treatment with different concentrations of ciproxifan per 300 μm. Data are presented as mean ± SEM. Statistical analysis was performed using one-way ANOVA followed by Bonferroni’s test; ** p < 0.01. NF200 (green), phalloidin (red), DAPI (blue); n = 6 per group ( n = 24 in total). Scale bars: 50 μm in a 1–3 , b 1–3 , and c 1–3 ; 20 μm in a 4 , b 4 , and c 4

Article Snippet: Nuclear staining was performed using ProLong Gold Antifade Mountant with DAPI (cat. #8961S, Cell Signaling Technology, USA).

Techniques: Immunofluorescence