Expression profiling of cochlear genes uncovers sex-based cellular function in mouse cochleae.

Sex is a pivotal biological factor that significantly impacts tissue homeostasis and disease susceptibility. In the auditory system, sex differences have been observed in cochlear physiology and responses to pathological conditions. However, the underlying molecular mechanisms responsible for these differences remain elusive. The current research explores the differences in gene expression profiles in the cochlea between male and female mice, aiming to understand the functional implication of sex-biased gene expression in each sex. Using RNA-sequencing analysis on cochlear tissues obtained from male and female mice, we identified a significant number of genes exhibiting sex-biased expression differences. While some of these differentially expressed genes are located on sex chromosomes, most are found on autosomal chromosomes. Further bioinformatic analysis revealed that these genes are involved in several key cellular functions. In males, these genes are notably linked to oxidative phosphorylation and RNA synthesis and processing, suggesting their involvement in mitochondrial energy production and regulatory control of gene expression. In contrast, sex-biased genes are associated with mechano-transduction and synaptic transmission within female cochleae. Collectively, our study provides valuable insights into the molecular differences between the sexes and emphasizes the need for future research to uncover their functional implications and relevance to auditory health and disease development.

Heterogeneity in macrophages along the cochlear spiral in mice: insights from SEM and functional analyses.

The susceptibility of sensory cells to pathological conditions differs between the apical and basal regions of the cochlea, and the cochlear immune system may contribute to this location-dependent variability. Our previous study found morphological differences in basilar membrane macrophages between the apical and basal regions of the cochlea. However, the details of this site-dependent difference and its underlying structural and biological basis are not fully understood. In this study, we utilized scanning electron microscopy to examine the ultrastructure of macrophages and their surrounding supporting structures. Additionally, we examined the phagocytic activities of macrophages and the expression of immune molecules in both apical and basal regions of the cochlea. We employed two mouse strains (C57BL/6J and B6.129P-Cx3cr1tm1Litt/J) and evaluated three experimental conditions: young normal (1-4 months), aging (11-19 months), and noise-induced damage (120 dB SPL for 1 h). Using scanning electron microscopy, we revealed location-specific differences in basilar membrane macrophage morphology and surface texture, architecture in mesothelial cell layers, and spatial correlation between macrophages and mesothelial cells in both young and older mice. Observations of macrophage phagocytic activities demonstrated that basal macrophages exhibited greater phagocytic activities in aging and noise-damaged ears. Furthermore, we identified differences in the expression of immune molecules between the apical and basal cochlear tissues of young mice. Finally, our study demonstrated that as the cochlea ages, macrophages in the apical and basal regions undergo a transformation in their morphologies, with apical macrophages acquiring certain basal macrophage features and vice versa. Overall, our findings demonstrate apical and basal differences in macrophage phenotypes and functionality, which are related to distinct immune and structural differences in the macrophage surrounding tissues.

Galectin-3 protects auditory function in female mice.

Sex differences in the development of sensorineural hearing loss have been recognized in various inner ear disorders, but the molecular basis for such differences is poorly understood. Autosomal genes have been shown to cause sex differences in disease susceptibility, but many genes exerting sex-dependent effects on auditory function remain to be identified. Galectin-3 (Gal-3), a protein encoded by the autosomal gene Lgals3, is a member of the ß-galactoside-binding protein family, and has been linked to multiple biological processes, including immune responses, apoptosis, and cell adhesion. Here, we investigated auditory function and hair cell integrity in Gal-3 knockout (KO, Lgals3-/-) and wild-type (WT, Lgals3+/+) mice from age 1 to 6 months. KO mice show a more rapid age-related increase in ABR thresholds compared to WT mice. Noticeably, the threshold deterioration in female KO mice is significantly greater than in the male KO and WT mice. The ABR threshold elevation manifests over a broad frequency range in female KO mice, whereas the threshold elevations are confined to high frequencies in the male KO and WT mice. Moreover, DPOAE input/output functions reveal a similar pattern of auditory dysfunction, with the female KO mice displaying a significantly greater reduction in DPOAE amplitudes than male KO mice and WT mice of both sexes. Finally, age-related outer hair cell loss is greater for female KO mice compared to male KO mice and WT mice of both sexes. Together, these results indicate that Gal-3 deficiency exacerbates age-related cochlear degeneration and auditory dysfunction in female mice. Our study identifies Gal-3 as a sex-dependent molecule for maintaining female cochlear integrity.

The expression of VEGF and VEGFR in endotoxin induced otitis media with effusion in rats.

OBJECTIVE: To investigate the expression and correlation of vascular endothelial growth factor (VEGF) and its receptor with hypoxia-inducible factor-1 α (HIF-1 α) in otitis media with effusion (OME). METHODS: A rat model of OME was induced by injection of lipopolysaccharide (LPS) into the middle ear. Hematoxylin and eosin (HE) staining was used to observe the pathomorphological changes of the tympanic cavity in the middle ear of rats. Immunohistochemistry (IHC), western blotting and RT-qPCR were used to determine the mRNA and protein expression of VEGF, VEGFR-1, VEGFR-2 and HIF-1α in mucosa of tympanic cavity mucosa, respectively. RESULTS: In the OME group, the epithelial space of the middle ear mucosa was significantly thickened and infiltration of a large number of inflammatory cells was found on postoperative day (POD), and the otitis media basically subsided 2 weeks after operation. VEGF mRNA expression was significantly increased on POD 1, and its protein expression peaked on POD 3. HIF-1α mRNA expression was significantly increased and peaked on POD 1, while its protein expression began to increase on POD 3 and was significantly expressed in the middle ear mucosal epithelium. HIF-1α mRNA showed a positive correlation with VEGF mRNA and VEGFR-1 mRNA expression. CONCLUSION: VEGF mainly plays a role in the acute phase of OME, and it is abundantly expressed mediated by HIF-1α. And then it play a role in vasodilatation and increase of vascular permeability, thus promoting the generation of middle ear effusion.

Effect of vascular endothelial growth factor and its receptors in adult otitis media with effusion.

PURPOSE: Some studies have demonstrated that vascular endothelial growth factor (VEGF) plays a critical role in the pathogenesis of otitis media with effusion (OME) in animal models. However, the levels of VEGF and its receptors in adult OME have not been clarified. Our study was designed to detect the levels of VEGF and its receptors in adult OME and explore their relationship with effusion types, duration and prognosis of OME. METHODS: 61 patients with secretory otitis media were enrolled including 21 males and 40 females, with an average age of 54.7 ± 17.5 years. The middle-ear effusions were collected by tympanocentesis or myringotomy. The protein concentrations were determined by enzyme-linked immunosorbent assay and messenger RNA by real-time quantitative PCR. RESULTS: VEGF level was higher in AOME group, but not correlated with the recurrence of OME. VEGFR1 and VEGFR2 levels were lower in recurrent group compared with non-recurrent group. VEGFR2 level was higher in serous effusions than mucoid effusions. VEGF messenger RNA was positively correlated both with HIF-1α and MUC5B. CONCLUSIONS: VEGF and its receptors function to induce the production of middle-ear effusions (MEEs) at acute stage of OME rather than chronic or recurrent stage, which is mainly mediated by HIF-1α pathway. The formation of mucoid effusions is associated with MUC5B and VEGFR2, but not with duration and recurrence of OME.