Ear Development Research Articles

The RHW1-ZCN4 regulatory pathway confers natural variation of husk leaf width in maize.

Maize husk leaf - the outer leafy layers covering the ear - modulates kernel yield and quality. Despite its importance, however, the genetic controls underlying husk leaf development remain elusive. Our previous genome-wide association study identified a single nucleotide polymorphism located in the gene RHW1 (Regulator of Husk leaf Width) that is significantly associated with husk leaf-width diversity in maize. Here, we further demonstrate that a polymorphic 18-bp InDel (insertion/deletion) variant in the 3' untranslated region of RHW1 alters its protein abundance and accounts for husk leaf width variation. RHW1 encodes a putative MYB-like transcriptional repressor. Disruption of RHW1 altered cell proliferation and resulted in a narrower husk leaf, whereas RHW1 overexpression yielded a wider husk leaf. RHW1 positively regulated the expression of ZCN4, a well-known TFL1-like protein involved in maize ear development. Dysfunction of ZCN4 reduced husk leaf width even in the context of RHW1 overexpression. The InDel variant in RHW1 is subject to selection and is associated with maize husk leaf adaption from tropical to temperate regions. Overall, our results identify that RHW1-ZCN4 regulates a pathway conferring husk leaf width variation at a very early stage of husk leaf development in maize.

Lmo4a Contributes to Zebrafish Inner Ear and Vestibular Development via Regulation of the Bmp Pathway.

In vertebrates, the development of the inner ear is a delicate process, whereas its relating molecular pathways are still poorly understood. LMO4, an LIM domain-only transcriptional regulator, is drawing an increasing amount of interest for its multiple roles regarding human embryonic development and the modulation of ototoxic side effects of cisplatin including cochlear apoptosis and hearing loss. The aim of the present study is to further explore the role of lmo4a in zebrafish inner ear development and thus explore its functional role. The Spatial Transcript Omics DataBase was referred to in order to evaluate the expression of lmo4a during the first 24 h of zebrafish development. In situ hybridization was applied to validate and extend the expression profile of lmo4a to 3 days post-fertilization. The morpholino (MO) knockdown and CRISPR/Cas9 knockout (KO) of lmo4a was applied. Morphological analyses of otic vesical, hair cells, statoacoustic ganglion and semicircular canals were conducted. The swimming pattern of lmo4a KO and MO zebrafish was tracked. In situ hybridization was further applied to verify the expression of genes of the related pathways. Rescue of the phenotype was attempted by blockage of the bmp pathway via heat shock and injection of Dorsomorphin. lmo4a is constitutively expressed in the otic placode and otic vesicle during the early stages of zebrafish development. Knockdown and knockout of lmo4a both induced smaller otocysts, less hair cells, immature statoacoustic ganglion and malformed semicircular canals. Abnormal swimming patterns could be observed in both lmo4a MO and KO zebrafish. eya1 in preplacodal ectoderm patterning was downregulated. bmp2 and bmp4 expressions were found to be upregulated and extended in lmo4a morphants, and blockage of the Bmp pathway partially rescued the vestibular defects. We concluded that lmo4a holds a regulative effect on the Bmp pathway and is required for the normal development of zebrafish inner ear. Our study pointed out the conservatism of LMO4 in inner ear development between mammals and zebrafish as well as shed more light on the molecular mechanisms behind it. Further research is needed to distinguish the relationships between lmo4 and the Bmp pathway, which may lead to diagnostic and therapeutic approaches towards human inner ear malformation.

Chicken utricle stromal cell-derived decellularized extracellular matrix coated nanofibrous scaffolds promote auditory cell production

Stem cell therapy has a broad future in treating sensorineural hearing loss in mammals. But how to produce sufficient functional auditory cells including hair cells, supporting cells as well as spiral ganglion neurons from potential stem cells is the bottleneck. In this study, we aimed to simulate inner ear development microenvironment to induce inner ear stem cells to differentiate into auditory cells. The different mass ratios of poly-l-lactic acid/gelatin (PLLA/Gel) scaffolds were fabricated by electrospinning technology to mimic the structure of the native cochlear sensory epithelium. The chicken utricle stromal cells were isolated and cultured, and then seeded on the PLLA/Gel scaffolds. The chicken utricle stromal cell-derived decellularized extracellular matrix (U-dECM)-coated PLLA/Gel bioactive nanofiber scaffolds (U-dECM/PLLA/Gel) were prepared by decellularization. The U-dECM/PLLA/Gel scaffolds were used for culture of inner ear stem cells, and the effects of the modified scaffolds on the differentiation of inner ear stem cells were analyzed by RT-PCR and immunofluorescent staining. The results showed that U-dECM/PLLA/Gel scaffolds possessed good biomechanical properties can significantly promote the differentiation of inner ear stem cells and make them differentiate into auditory cells. Collectively, these findings indicated that U-dECM-coated biomimetic nanomaterials may be a promising strategy for auditory cell production.

The spontaneous mouse mutant low set ears (Lse) is caused by tandem duplication of Fgf3 and Fgf4.

The external ear develops from an organized convergence of ventrally migrating neural crest cells into the first and second branchial arches. Defects in external ear position are often symptomatic of complex syndromes such as Apert, Treacher-Collins, and Crouzon Syndrome. The low set ears (Lse) spontaneous mouse mutant is characterized by the dominant inheritance of a ventrally shifted external ear position and an abnormal external auditory meatus (EAM). We identified the causative mutation as a 148Kb tandem duplication on Chromosome 7, which includes the entire coding sequences of Fgf3 and Fgf4. Duplications of FGF3 and FGF4 occur in 11q duplication syndrome in humans and are associated with craniofacial anomalies, among other features. Intercrosses of Lse-affected mice revealed perinatal lethality in homozygotes, and Lse/Lse embryos display additional phenotypes including polydactyly, abnormal eye morphology, and cleft secondary palate. The duplication results in increased Fgf3 and Fgf4 expression in the branchial arches and additional discrete domains in the developing embryo. This ectopic overexpression resulted in functional FGF signaling, demonstrated by increased Spry2 and Etv5 expression in overlapping domains of the developing arches. Finally, a genetic interaction between Fgf3/4 overexpression and Twist1, a regulator of skull suture development, resulted in perinatal lethality, cleft palate, and polydactyly in compound heterozygotes. These data indicate a role for Fgf3 and Fgf4 in external ear and palate development and provide a novel mouse model for further interrogation of the biological consequences of human FGF3/4 duplication.

Isolated Aplasia of the Lateral Semicircular Canal without Hearing Loss: A Case Report

We present a case of a 25-year-old woman with isolated aplasia of the lateral semicircular canal (LSCC), a rare congenital malformation that results in the absence or underdevelopment of the LSCC. The patient had vertigo, left-sided tinnitus, and persistent chronic headaches but no hearing loss. High-resolution computed tomography (CT) and magnetic resonance imaging (MRI) were used to detect the malformation and associated vasculo-neural conflict. The aplasia of the LSCC was the only abnormality observed. The article explains the embryology of inner ear development and associated anomalies. CT and MRI were found to be effective imaging techniques for detecting ear malformations. The article also explains the importance of proper diagnosis and assessment of hearing loss for patients with this malformation.

Defining developmental trajectories of prosensory cells in human inner ear organoids at single-cell resolution.

The inner ear sensory epithelia contain mechanosensitive hair cells and supporting cells. Both cell types arise from SOX2-expressing prosensory cells, but the mechanisms underlying the diversification of these cell lineages remain unclear. To determine the transcriptional trajectory of prosensory cells, we established a SOX2-2A-ntdTomato human embryonic stem cell line using CRISPR/Cas9, and performed single-cell RNA-sequencing analyses with SOX2-positive cells isolated from inner ear organoids at various time points between differentiation days 20 and 60. Our pseudotime analysis suggests that vestibular type II hair cells arise primarily from supporting cells, rather than bi-fated prosensory cells in organoids. Moreover, ion channel- and ion-transporter-related gene sets were enriched in supporting cells vs. prosensory cells, whereas Wnt signaling-related gene sets were enriched in hair cells vs. supporting cells. These findings provide valuable insights into how prosensory cells give rise to hair cells and supporting cells during human inner ear development, and may provide a clue to promote hair cell regeneration from resident supporting cells in individuals with hearing loss or balance disorders.

Graphene Hybrid Inner Ear Organoid with Enhanced Maturity.

Inner ear organoids (IEOs) are 3D structures grown in vitro, which can mimic the complex cellular structure and function of the inner ear. IEOs are potential solutions to problems related to inner ear development, disease modeling, and drug delivery. However, current approaches in generating IEOs using chemical factors have a few limitations, resulting in unpredictable outcomes. In this study, we propose the use of nanomaterial-based approaches, specifically by using graphene oxide (GO). GO's unique properties promote cell-extracellular matrix interactions and cell-cell gap junctions, thereby enhancing hair cell formation, which is an essential part of IEO development. We also investigated the potential applications for drug testing. Our findings suggest that GO is a promising candidate for enhancing the functionality of IEOs and advancing our understanding of the problems underlying inner ear development. The use of nanomaterial-based approaches may provide a more reliable and effective method for building better IEOs in the future.

A clinical case of a patient with an anomaly in the development of the inner ear and refractory stimulation of the facial nerve after a coronavirus infection and meningitis

Background. Thanks to the development of technology and new surgical approaches, it has become possible to perform cochlear implantation (CI) in patients with malformations of the inner ear. The experience of this type of rehabilitation has more than 15 years in Russian practice and more than 20 years in the world. Unfortunately, the COVID-19 pandemic has added not only one more cause of deafness, but also etiological factor of the complications. The purpose of this paper is to describe a clinical case of facial nerve stimulation (FNS) in a CI user with an inner ear malformation and a coronavirus infection complicated by meningitis. A clinical case demonstrating a combination of all these etiological factors of FNS has not been previously described in the scientific literature.
 Aim. To describe a clinical case of FNS in a CI user with an inner ear malformation and a coronavirus infection complicated by meningitis.
 Materials and methods. In 2017 4 years old patient underwent cochlear implantation on her right ear in Saint Petersburg Research Institute of Ear, Throat, Nose and Speech. In May 2022 the patient was admitted to the infectious diseases department of the medical institution with the diagnosis: coronavirus infection (confirmed), severe course; complication: purulent meningoencephalitis. After recovery, the patient complained of facial muscles twitching when using the speech processor of the cochlear implant. In order to correct the non-hearing sensation, the patient was sent to the Saint Petersburg Research Institute of Ear, Throat, Nose and Speech for rehabilitation in June 2022. During repeated fitting, it was possible to improve the reactions to sounds. Phoneme discrimination has improved. Positive dynamics was registered against the background of the changed settings. The average hearing thresholds using 3-phase stimulation were 42.5 dB. However, it was not possible to completely eliminate the FNS, and the compulsory settings of the speech processor did not allow the patient to achieve sufficient speech intelligibility.
 Conclusion. Meningitis after CI can cause an increase in the transmittance of the inner ear tissues to electric current and, as a consequence, the development of the FNS. Modern technologies make it possible to rehabilitate such patients, including those with abnormalities of the inner ear. To increase the effecacy of rehabilitation, special coding strategies are used including the 3-phase stimulation.

Thyroid Axis and Vestibular Physiopathology: From Animal Model to Pathology.

A recent work of our group has shown the significant effects of thyroxine treatment on the restoration of postural balance function in a rodent model of acute peripheral vestibulopathy. Based on these findings, we attempt to shed light in this review on the interaction between the hypothalamic-pituitary-thyroid axis and the vestibular system in normal and pathological situations. Pubmed database and relevant websites were searched from inception through to 4 February 2023. All studies relevant to each subsection of this review have been included. After describing the role of thyroid hormones in the development of the inner ear, we investigated the possible link between the thyroid axis and the vestibular system in normal and pathological conditions. The mechanisms and cellular sites of action of thyroid hormones on animal models of vestibulopathy are postulated and therapeutic options are proposed. In view of their pleiotropic action, thyroid hormones represent a target of choice to promote vestibular compensation at different levels. However, very few studies have investigated the relationship between thyroid hormones and the vestibular system. It seems then important to more extensively investigate the link between the endocrine system and the vestibule in order to better understand the vestibular physiopathology and to find new therapeutic leads.

Transcriptome analysis reveals an Atoh1b-dependent gene set downstream of Dlx3b/4b during early inner ear development in zebrafish.

The vertebrate inner ear is the sensory organ mediating hearing and balance. The entire organ develops from the otic placode, which itself originates from the otic-epibranchial progenitor domain (OEPD). Multiple studies in various species have shown the importance of the forkhead-box and distal-less homeodomain transcription factor families for OEPD and subsequent otic placode formation. However, the transcriptional networks downstream of these factors are only beginning to be understood. Using transcriptome analysis, we here reveal numerous genes regulated by the distal-less homeodomain transcription factors Dlx3b and Dlx4b (Dlx3b/4b). We identify known and novel transcripts displaying widespread OEPD expression in a Dlx3b/4b-dependent manner. Some genes, with a known OEPD expression in other vertebrate species, might be members of a presumptive vertebrate core module required for proper otic development. Moreover, we identify genes controlling early-born sensory hair cell formation as well as regulating biomineral tissue development, both consistent with defective sensory hair cell and otolith formation observed in dlx3b/4b mutants. Finally, we show that ectopic Atoh1b expression can rescue early sensorigenesis even in the absence of Dlx3b/4b. Taken together, our data will help to unravel the gene regulatory network underlying early inner ear development and provide insights into the molecular control of vertebrate inner ear formation to restore hearing loss in humans ultimately.

A single-cell level comparison of human inner ear organoids with the human cochlea and vestibular organs

Inner ear disorders are among the most common congenital abnormalities; however, current tissue culture models lack the cell type diversity to study these disorders and normal otic development. Here, we demonstrate the robustness of human pluripotent stem cell-derived inner ear organoids (IEOs) and evaluate cell type heterogeneity by single-cell transcriptomics. To validate our findings, we construct a single-cell atlas of human fetal and adult inner ear tissue. Our study identifies various cell types in the IEOs including periotic mesenchyme, type I and type II vestibular hair cells, and developing vestibular and cochlear epithelium. Many genes linked to congenital inner ear dysfunction are confirmed to be expressed in these cell types. Additional cell-cell communication analysis within IEOs and fetal tissue highlights the role of endothelial cells on the developing sensory epithelium. These findings provide insights into this organoid model and its potential applications in studying inner ear development and disorders.

Abnormal ear development in corn: Does hybrid, environment, and seeding rate matter?

AbstractCorn (Zea mays L.) yields have increased in the United States since the 1930s and in other parts of the world since the 1950s and 1960s because of improvements in agricultural management and genotypes. Despite these increases, production concerns still exist. In July 2016, abnormal ear development (multi‐ears per node, barbell‐ears, and short‐husks) was reported in cornfields that extended from the Texas Panhandle to eastern Colorado and east through Kansas, Nebraska, Iowa, and Illinois. Surveys in Nebraska farmer fields revealed significant productivity losses due to the issues, but little was known about the underlying causes. A research study was conducted in four Nebraska fields during the 2018 and 2019 growing seasons. The research investigated the effects of hybrids, environments, seeding rates, and their interactions on abnormal ears. Eight hybrids, eight environments, and five seeding rates were studied. About 63,500 plants were individually assessed at or after the dent stage (R5). Grain yield ranged from 4.3 to 20.1 Mg ha−1. In 2018, about 5% of ears were abnormal; in 2019, about 11%, if combined, about 8%. Higher‐yielding hybrids were associated with lower percentages of abnormalities. Hybrids, environments, and seeding rates influenced the occurrence of abnormal ears. In most cases, abnormal ears had lower heights in the canopy, suggesting that primary ear loss may be a factor. The results reinforced the overriding hypothesis that ear abnormalities result from environmental, genetic, and management interactions. Depending on the environment, selecting certain hybrids with optimum seeding rates could help mitigate the occurrence of abnormal ears.

Dispensable role of Rac1 and Rac3 after cochlear hair cell specification

Rac small GTPases play important roles during embryonic development of the inner ear; however, little is known regarding their function in cochlear hair cells (HCs) after specification. Here, we revealed the localization and activation of Racs in cochlear HCs using GFP-tagged Rac plasmids and transgenic mice expressing a Rac1-fluorescence resonance energy transfer (FRET) biosensor. Furthermore, we employed Rac1-knockout (Rac1-KO, Atoh1-Cre;Rac1flox/flox) and Rac1 and Rac3 double KO (Rac1/Rac3-DKO, Atoh1-Cre;Rac1flox/flox;Rac3−/−) mice, under the control of the Atoh1 promoter. However, both Rac1-KO and Rac1/Rac3-DKO mice exhibited normal cochlear HC morphology at 13 weeks of age and normal hearing function at 24 weeks of age. No hearing vulnerability was observed in young adult (6-week-old) Rac1/Rac3-DKO mice even after intense noise exposure. Consistent with prior reports, the results from Atoh1-Cre;tdTomato mice confirmed that the Atoh1 promoter became functional only after embryonic day 14 when the sensory HC precursors exit the cell cycle. Taken together, these findings indicate that although Rac1 and Rac3 contribute to the early development of sensory epithelia in cochleae, as previously shown, they are dispensable for the maturation of cochlear HCs in the postmitotic state or for hearing maintenance following HC maturation.Key messagesMice with Rac1 and Rac3 deletion were generated after HC specification.Knockout mice exhibit normal cochlear hair cell morphology and hearing.Racs are dispensable for hair cells in the postmitotic state after specification.Racs are dispensable for hearing maintenance after HC maturation.

An Increase in Solar Radiation in the Late Growth Period of Maize Alleviates the Adverse Effects of Climate Warming on the Growth and Development of Maize

Against the background of long-term climate change, quantifying the response of maize growth and development to climate change during critical growth stages will contribute to future decision-making in maize production. However, there have been few reports on the impact of climate change on maize dry matter accumulation and yield formation using long-term field trial data. In this study, field trial data from 13 agricultural meteorological stations in the Beijing–Tianjin–Hebei region from 1981 to 2017 were analyzed using partial correlation analysis and multiple regression models to investigate the effects of climate change on maize growth and yield. The results showed that the average temperature (Tavg) and accumulated effective precipitation (EP) during the maize growing season increased while the accumulated solar radiation (SRD) decreased from 1981 to 2017. During the seedling stage (GS1, VE-V8) and ear development stage (GS2, V8-R1), Tavg increased by 0.34 °C and 0.36 °C/decade, respectively, and EP increased by 1.83 mm and 3.35 mm/decade, respectively. The significant increase in Tavg during GS1 was the main reason for the inhibitory effect of climate change on maize growth, development, and biomass accumulation. However, the increase in SRD during the grain formation stage (GS3, R1–R3) and grain filling stage (GS4, R3–R6) was favorable for yield formation, increasing the grain number per ear (GN) and grain weight (GW) by 5.00% and 2.84%, respectively. SRD significantly increased after the silk stage, partially offsetting the adverse effects of temperature on maize yield formation, but the final result was a 0.18% and 0.94% reduction in maize plant dry weight (TDW) and grain yield (GY), respectively, due to the combined effects of the three climate factors. Spatially, climate change mainly had a negative impact on maize in the eastern and western parts of the central region of Beijing–Tianjin–Hebei, with a maximum GY reduction of up to 34.06%. The results of this study can provide a scientific basis for future decision-making in maize production against the background of climate change.

Prkra Mutation Alters mRNA Expression During Embryonic External Ear Development.

To understand the changes in mRNA expression during the embryonic development of the external mouse ear after the point mutation of the Prkra gene, Prkra short ear mouse model was used to study the development of the embryonic external ear. The tissues of the embryonic external ear were obtained when mouse embryos developed to E15.5 and E17.5. The changes in the mRNA expression profile were detected and analyzed. Find_circ and CIRI2 softwares were used to identify the upregulated and down-regulated expression of mRNA in the experimental and control groups. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes functional annotations were conducted on the differentially expressed mRNA, and the related signal pathways were analyzed after the upregulation and down-regulation of mRNA expression. This study aimed to understand the regulation of mRNA expression in Prkra short-ear mice during the external ear development in embryos. The results showed a correlation between abnormally expressed mRNA and signal pathways and the regulation of the development of the external ear of Prkra short-ear mice, and there were differences in some key regulatory mRNA changes after the Prkra gene point mutation. This study will provide a new clue for the mechanism of mRNA regulating the development of the external mouse ear. The change in mRNA expression profile can also provide clues for studying the biological regulation mechanism of external ear embryonic development.

A dominant variant in apoptosis-related gene XKR8 is relevant to hereditary auditory neuropathy

BackgroundAuditory neuropathy is an unusual type of hearing loss. At least 40% of patients with this disease have underlying genetic causes. However, in many hereditary auditory neuropathy cases, etiology remains undetermined.MethodsWe collected data and blood samples from a four-generation Chinese family. After excluding relevant variants in known deafness-related genes, exome sequencing was conducted. Candidate genes were verified by pedigree segregation, transcript/protein expression in the mouse cochlea, and plasmid expression studies in HEK 293T cells. Moreover, a mutant mouse model was generated and underwent hearing evaluations; protein localization in the inner ear was also assessed.ResultsThe clinical features of the family were diagnosed as auditory neuropathy. A novel variant c.710G > A (p.W237X) in apoptosis-related gene XKR8 was identified. Genotyping of 16 family members confirmed the segregation of this variant with the deafness phenotype. Both XKR8 mRNA and XKR8 protein were expressed in the mouse inner ear, predominantly in regions of spiral ganglion neurons; Moreover, this nonsense variant impaired the surface localization of XKR8 in cells. Transgenic mutant mice exhibited late-onset auditory neuropathy, and their altered XKR8 protein localization in the inner ear confirmed the damaging effects of this variant.ConclusionsWe identified a variant in the XKR8 gene that is relevant to auditory neuropathy. The essential role of XKR8 in inner ear development and neural homeostasis should be explored.

Development of otic organoids and their current status

The inner ear is responsible for both hearing and balance in the body, and since the initial development of otic (inner ear) organoids from mouse pluripotent stem cells (PSCs) in 2013, significant advances have been made in this field. Bone morphogenetic proteins, fibroblast growth factors, and Wnt agonists, which are signaling molecules in the early development of the inner ear, can induce PSCs into the otic fate. In the inner ear, hair cells and the surrounding supporting cells are essential for proper function and structure. Recent advancements in otic organoid research have enabled the generation of cells that closely resemble these key components. The developed otic organoids contain both hair cell-like cells and supporting cells, which have been confirmed to have the intrinsic function of those cell types. Otic organoids have been used for disease modeling and are expected to be more widely applied in various areas of research on the inner ear. However, the otic organoids developed to date remain immature. Although they mimic hair cells, their properties resemble vestibular (balance) hair cells more closely than cochlear (auditory) hair cells. The ultimate goal of research on the inner ear is hearing restoration and prevention; thus, it is essential to produce otic organoids that contain cochlear hair cells. In addition, the organ of Corti—a cell arrangement unique to the cochlea—has not yet been simulated. Along with a description of the current status of otic organoids, this review article will discuss future directions for otic organoids in inner ear research.

Involvement of Dmp1 in the Precise Regulation of Hair Bundle Formation in the Developing Cochlea.

Dentin matrix protein 1 (Dmp1) is a highly phosphorylated, extracellular matrix protein that is extensively expressed in bone and teeth but also found in soft tissues, including brain and muscle. However, the functions of Dmp1 in the mice cochlea are unknown. Our study showed that Dmp1 was expressed in auditory hair cells (HCs), with the role of Dmp1 in those cells identified using Dmp1 cKD mice. Immunostaining and scanning electron microscopy of the cochlea at P1 revealed that Dmp1 deficiency in mice resulted in an abnormal stereociliary bundle morphology and the mispositioning of the kinocilium. The following experiments further demonstrated that the cell-intrinsic polarity of HCs was affected without apparent effect on the tissue planer polarity, based on the observation that the asymmetric distribution of Vangl2 was unchanged whereas the Gαi3 expression domain was enlarged and Par6b expression was slightly altered. Then, the possible molecular mechanisms of Dmp1 involvement in inner ear development were explored via RNA-seq analysis. The study suggested that the Fgf23-Klotho endocrine axis may play a novel role in the inner ear and Dmp1 may regulate the kinocilium-stereocilia interaction via Fgf23-Klotho signaling. Together, our results proved the critical role of Dmp1 in the precise regulation of hair bundle morphogenesis in the early development of HCs.

Exposure to the persistent organic pollutant 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD, dioxin) disrupts development of the zebrafish inner ear

Dioxins are a class of highly toxic and persistent environmental pollutants that have been shown through epidemiological and laboratory-based studies to act as developmental teratogens. 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), the most potent dioxin congener, has a high affinity for the aryl hydrocarbon receptor (AHR), a ligand activated transcription factor. TCDD-induced AHR activation during development impairs nervous system, cardiac, and craniofacial development. Despite the robust phenotypes previously reported, the characterization of developmental malformations and our understanding of the molecular targets mediating TCDD-induced developmental toxicity remains limited. In zebrafish, TCDD-induced craniofacial malformations are produced, in part, by the downregulation of SRY-box transcription factor 9b (sox9b), a member of the SoxE gene family. sox9b, along with fellow SoxE gene family members sox9a and sox10, have important functions in the development of the otic placode, the otic vesicle, and, ultimately, the inner ear. Given that sox9b is a known target of TCDD and that transcriptional interactions exist among SoxE genes, we asked whether TCDD exposure impaired the development of the zebrafish auditory system, specifically the otic vesicle, which gives rise to the sensory components of the inner ear. Using immunohistochemistry, in vivo confocal imaging, and time-lapse microscopy, we assessed the impact of TCDD exposure on zebrafish otic vesicle development. We found exposure resulted in structural deficits, including incomplete pillar fusion and altered pillar topography, leading to defective semicircular canal development. The observed structural deficits were accompanied by reduced collagen type II expression in the ear. Together, our findings reveal the otic vesicle as a novel target of TCDD-induced toxicity, suggest that the function of multiple SoxE genes may be affected by TCDD exposure, and provide insight into how environmental contaminants contribute to congenital malformations.

Early Steps towards Hearing: Placodes and Sensory Development.

Sensorineural hearing loss is the most prevalent sensory deficit in humans. Most cases of hearing loss are due to the degeneration of key structures of the sensory pathway in the cochlea, such as the sensory hair cells, the primary auditory neurons, and their synaptic connection to the hair cells. Different cell-based strategies to replace damaged inner ear neurosensory tissue aiming at the restoration of regeneration or functional recovery are currently the subject of intensive research. Most of these cell-based treatment approaches require experimental in vitro models that rely on a fine understanding of the earliest morphogenetic steps that underlie the in vivo development of the inner ear since its initial induction from a common otic-epibranchial territory. This knowledge will be applied to various proposed experimental cell replacement strategies to either address the feasibility or identify novel therapeutic options for sensorineural hearing loss. In this review, we describe how ear and epibranchial placode development can be recapitulated by focusing on the cellular transformations that occur as the inner ear is converted from a thickening of the surface ectoderm next to the hindbrain known as the otic placode to an otocyst embedded in the head mesenchyme. Finally, we will highlight otic and epibranchial placode development and morphogenetic events towards progenitors of the inner ear and their neurosensory cell derivatives.