Utricular Macula Research Articles

Membrana limitans shape variation and its role in vestibular form

The membrana limitans (ML) is a membranous vestibular structure found within the bony labyrinth, housed in the petrous temporal bone. It is located inferior to the semicircular canals (which detect changes in angular acceleration of the head) and adjacent to both otolith organs, the utricle and saccule (responsible for detecting linear acceleration of the head and gravity). The ML forms the keystone to vestibular geometry, acting as a support for the majority of the utricular macula and respective neural substrate, and provides a structural boundary between the superior (pars superior) and inferior (pars inferior) portions of the vestibular labyrinth. Despite its importance, knowledge of ML 3D structure and morphological variation is limited. Here we test the hypothesis that there is consistency among ML bony attachment sites in humans. We use a sample comprised of micro-CT scans of nine human temporal bones soaked in 2 per cent osmium tetroxide after fixation in Karnovsky's fixative in order to better visualize the membranous labyrinth. Each ML was 3D reconstructed and the extent of bony attachment site variation was assessed using landmark-based Generalized Procrustes and Principal Component Analyses. Results indicate a consistent, yet locally variable configuration with distinct anterolateral, medial and posterior attachments of the ML to the bony wall of the labyrinth. The anterolateral attachment originates just superior to the posteriormost edge of the stapes footplate and stretches anteriorly. This is often a discontinuous attachment (observed in seven of nine ears) which occasionally attaches onto the posterior one-third of the footplate itself (observed in two of nine ears). The length of the medial attachment is the most variable, yet in all ears it attaches along the superior border of the spherical recess which houses the saccule. The posterior attachment is the least variable and its medial boundary forms a consistent attachment to the bony lip of the vestibular aqueduct (observed in all ears). These results also support previous findings suggesting the ML forms a discontinuous support system within the vestibule among humans. This exploratory analysis sets the stage for further understanding into utricular biomechanics and vestibular geometry, especially related to details of otolith organ structure and their relationships to bony anatomy.

Selective Asymmetry of Ocular Vestibular-Evoked Myogenic Potential in Patients with Acute Utricular Macula Loss.

We retrospectively evaluated a chart review of 3,525 patients evaluated for any acute disturbance. A total of 1,504 patients with acute vestibular syndrome (AVS) received an instrumental vestibular assessment within 72 h from the onset of the symptoms evaluated using simultaneously a combination of ocular vestibular-evoked myogenic potential (oVEMPs), cervical vestibular-evoked myogenic potential (cVEMPs), video head-impulse test (vHIT), and subjective visual vertical (SVV) were included in this study. A total of 41 patients with AVS that showed a normal horizontal canal function tested with vHIT, a normal cVEMP function, unilaterally reduced or absent oVEMP n10, and an altered SVV were enrolled. We found that although these patients referred acute vertigo and presented spontaneous nystagmus, they showed physiological values of vHIT and a normal saccular function, as shown by symmetrical cVEMPs. Our findings support the hypothesis that a percentage of patients evaluated during an AVS using an instrumental vestibular assessment could present selective utricular macula dysfunction.

Recurring paroxysmal positional vertigo: evaluation of the vascular factor.

SUMMARYTo evaluate the effective incidence of vascular factor in the recurrence of benign paroxysmal positional vertigo (BPPV), we studied 50 subjects, 32 affected by idiopathic recurrent BPPV (study group) and 18 healthy subjects (control group). All subjects underwent complete otoneurological balance and haemodynamic evaluation by extracranial colour-coded duplex sonography (ECCS) of vertebral arteries (VA) with indication of arterial flow in ml/min, and retinal fluorangiography (FAG). The ECCS of 19 patients (59.3%) within the study group presented a reduction in vertebral arterial flow, exceeding the limits established by normative values (< 100 ml/min). In all cases, the same side was affected by BPPV, emphasised by vertebral hypoperfusion. The remaining 13 patients (40.6%) showed an arterial vertebral flow entirely within the normative values. The FAG excluded qualitative alterations of the cerebral microcirculation. The ECCS demonstrated that 59.3% of the study group showed a significant reduction in vertebral arterial flow ipsilateral to the semicircular canal affected by BPPV. This increased to 68.75% when the flow difference (D) between both the vertebral arteries were considered and reached 71.8% when vascular risk factors were evaluated. We conclude that reduced perfusion of the vestibular structures makes an already critical situation even more difficult, which can eventually develop into labyrinth suffering. The absence of fluorangiographic signs suggests that the labyrinthine neuroepithelium is much more sensitive to hypoperfusion than the retina. We hypothesise that this ischaemic situation could degenerate utricular macula, otolith detachment, leading to the development of recurrent BPPV. This risk situation for the labyrinth can also be revealed by the evaluation of three parameters: presence of vascular risk factors, reduction of vertebral flow < 100 ml/min and the difference in flow between the 2 vertebral arteries > 29 ml/min.

Consistent removal of hair cells in vestibular end organs by time-dependent transtympanic administration of gentamicin in guinea pigs

BackgroundVestibular hair cell loss and its role in balance disorders are not yet completely understood due largely to the lack of precise hair cell damage protocols. New methodOur damage protocol aims to selectively remove type I hair cells in a way that produces consistent and predictable lesions that can be used for reliable inter-animal and inter-group comparison in balance research. This objective is achieved by transtympanic injection of gentamicin on both the round window membrane and oval window over a fixed time period followed by thorough washing. ResultsWe achieved nearly total and consistent loss of type I hair cells at 94 % for the crista ampullaris of the lateral semicircular canal (LSC) and 86 % for the utricular macula with negligible loss of type II hair cells at 4% for the crista ampullaris of the LSC and 6% for the utricular macula. While the vestibular function was compromised in the relevant study group, this group had a zero mortality rate with no significant suppression of body weight gain. Comparison with existing methodsGentamicin is typically administered via intraperitoneal systemic injection or, more recently, transtympanic injection. The intraperitoneal method is simple, but mortality rate is high. The transtympanic injection method produces ototoxic damage but with inconsistent lesion size. This inconsistency prevents reliable comparisons among animals. ConclusionsThis protocol employs a transtympanic injection method which selectively targets type I hair cells for removal in the vestibular epithelia in a time-dependent manner, uniformly damages vestibular function, and causes uniform hair cell loss.

The Anatomical and Physiological Basis of Clinical Tests of Otolith Function. A Tribute to Yoshio Uchino.

Otolithic receptors are stimulated by gravitoinertial force (GIF) acting on the otoconia resulting in deflections of the hair bundles of otolithic receptor hair cells. The GIF is the sum of gravitational force and the inertial force due to linear acceleration. The usual clinical and experimental tests of otolith function have used GIFs (roll tilts re gravity or linear accelerations) as test stimuli. However, the opposite polarization of receptors across each otolithic macula is puzzling since a GIF directed across the otolith macula will excite receptors on one side of the line of polarity reversal (LPR at the striola) and simultaneously act to silence receptors on the opposite side of the LPR. It would seem the two neural signals from the one otolith macula should cancel. In fact, Uchino showed that instead of canceling, the simultaneous stimulation of the oppositely polarized hair cells enhances the otolithic response to GIF—both in the saccular macula and the utricular macula. For the utricular system there is also commissural inhibitory interaction between the utricular maculae in each ear. The results are that the one GIF stimulus will cause direct excitation of utricular receptors in the activated sector in one ear as well as indirect excitation resulting from the disfacilitation of utricular receptors in the corresponding sector on the opposite labyrinth. There are effectively two complementary parallel otolithic afferent systems—the sustained system concerned with signaling low frequency GIF stimuli such as roll head tilts and the transient system which is activated by sound and vibration. Clinical tests of the sustained otolith system—such as ocular counterrolling to roll-tilt or tests using linear translation—do not show unilateral otolithic loss reliably, whereas tests of transient otolith function [vestibular evoked myogenic potentials (VEMPs) to brief sound and vibration stimuli] do show unilateral otolithic loss. The opposing sectors of the maculae also explain the results of galvanic vestibular stimulation (GVS) where bilateral mastoid galvanic stimulation causes ocular torsion position similar to the otolithic response to GIF. However, GVS stimulates canal afferents as well as otolithic afferents so the eye movement response is complex.

Association Between Benign Positional Vertigo and Migraine in Indian Population.

Benign positional vertigo (BPV) occurs when freely floating otoconia which are normally attached to the utricular macula, enter the posterior semicircular canal and move under the influence of gravity. It is the most common cause of peripheral vertigo. Migraine is a common headache disorder which is characterized by hemicranial, throbbing pain and may be preceded by aura. The relation between vertigo and migraine is intriguing and in day to day practice, often not clear. 100 diagnosed patients of BPV, aged more than 20years, were evaluated for presence of headache specifically migraine based on International Headache Classification 3rd Edition, beta version. We also compared the success of epley's manoeuvre in patients having BPV & headache as compared to patients with complaints of vertigo alone. Overall seventy-four patients had successful epley's manoeuver and the rest did not show improvement even after four trials. Thirty-four patients reported headache and migraine was diagnosed in only ten patients. 67.6% of patients with headache had successful epley's maneuver however fifty-one of the 66 patients without headache had successful epley's manoeuver. Prevalence of migraine was only 10% in patients with BPV in our population and we observed that presence of headache does not suggest success or failure of epley's manoeuvre.

Utricular Sensitivity during Hydrodynamic Displacements of the Macula.

To explore the effects of cochlear hair cell displacement, researchers have previously monitored functional and mechanical responses during low-frequency (LF) acoustic stimulation of the cochlea. The induced changes are believed to result from modulation of the conductance of mechano-electrical transduction (MET) channels on cochlear hair cells, along with receptor potential modulation. It is less clear how, or if, vestibular hair cell displacement affects vestibular function. Here, we have used LF (<20Hz) hydrodynamic modulation of the utricular macula position, whilst recording functional and mechanical responses, to investigate the effects of utricular macula displacement. Measured responses included the Utricular Microphonic (UM), the vestibular short-latency evoked potential (VsEP), and laser Doppler vibrometry recordings of macular position. Over 1cycle of the LF bias, the UM amplitude and waveform were cyclically modulated, with Boltzmann analysis suggesting a cyclic modulation of the vestibular MET gating. The VsEP amplitude was cyclically modulated throughout the LF bias, demonstrating a relative increase (~20-50%; re baseline) and decrease (~10-20%; re baseline), which is believed to be related to the MET conductance and vestibular hair cell sensitivity. The relationship between macular displacement and changes in UM and VsEP responses was consistent within and across animals. These results suggest that the sensory structures underlying the VsEP, often thought to be a cranial jerk-sensitive response, are at least partially sensitive to LF (and possibly static) pressures or motion. Furthermore, these results highlight the possibility that some of the vestibular dysfunction related to endolymphatic hydrops may be due to altered vestibular transduction following mechanical (or morphological) changes in the labyrinth.

Constant severe imbalance following traumatic otoconial loss: a new explanation of residual dizziness.

Benign paroxysmal positional vertigo (BPPV) is the most common type of vertigo, caused by otoconia falling from the utricle into a semicircular canal (SCC). After successful repositioning maneuvers residual dizziness (RD) has been described and several reasons are used to explain RD. It can last for only a few days or weeks, but also much longer. We present a patient with a severe traumatic loss of otoconia from both maculae utriculi and a persistent imbalance more than 9years. We think that the loss of otoconia from the utricular and probably also saccular macula induced a sudden reduction of her ability to sense gravity thus logically explaining her symptoms. We show the vestibular test results also supporting our hypothesis and we extrapolate this support to other forms of so far unexplained dizziness especially increasing imbalance with aging. We also discuss the normal c- and oVEMP indicating intact haircell function and supporting our hypothesis of isolated otoconial loss as the major cause for imbalance.

Review of the pathology underlying benign paroxysmal positional vertigo.

The pathophysiological mechanism underlying benign paroxysmal positional vertigo (BPPV) is related to free-floating debris/otoliths in the semicircular canal (canalolithiasis) or debris/otoliths attached to the cupula (cupulolithiasis). These debris/otoliths are considered to originally accumulate after detachment from the neuroepithelium of the utricular macula secondary to a type of degeneration. An idiopathic form, which is assumed to occur spontaneously, is diagnosed when the causative pathology is obscure. However, an association between various other systemic or inner ear conditions and BPPV has been reported, indicating the existence of secondary BPPV. This study was performed to present the first review of the pathology underlying BPPV following a complete PubMed/Medline search. In total, 1932 articles published from 1975 to 2018 were reviewed. The articles were classified according to 17 potentially causative factors (aging; migraine; Meniere’s disease; infection; trauma; idiopathic sudden sensorineural hearing loss; sleeping habits; osteoporosis and vitamin D insufficiency; hyperglycemia and diabetes mellitus; chronic head and neck pain; vestibule or semicircular canal pathology; pigmentation disorders; estrogen deficiency; neurological disorders; autoimmune, inflammatory, or rheumatologic disorders; familial or genetic predisposition; and allergy). A discussion of the underlying cause of BPPV for each factor is presented.

Transcriptomic Analyses of Inner Ear Sensory Epithelia in Zebrafish.

Analysis of gene expression has the potential to assist in the understanding of multiple cellular processes including proliferation, cell-fate specification, senesence, and activity in both healthy and disease states. Zebrafish model has been increasingly used to understand the process of hearing and the development of the vertebrate auditory system. Within the zebrafish inner ear, there are three otolith organs, each containing a sensory macula of hair cells. The saccular macula is primarily involved in hearing, the utricular macula is primarily involved in balance and the function of the lagenar macula is not completely understood. The goal of this study is to understand the transcriptional differences in the sensory macula associated with different otolith organs with the intention of understanding the genetic mechanisms responsible for the distinct role each organ plays in sensory perception. The sensory maculae of the saccule, utricle, and lagena were dissected out of adult Et(krt4:GFP)sqet4 zebrafish expressing green fluorescent protein in hair cells for transcriptional analysis. The total RNAs of the maculae were isolated and analyzed by RNA GeneChip microarray. Several of the differentially expressed genes are known to be involved in deafness, otolith development and balance. Gene expression among these otolith organs was very well conserved with less than 10% of genes showing differential expression. Data from this study will help to elucidate which genes are involved in hearing and balance. Furthermore, the findings of this study will assist in the development of the zebrafish model for human hearing and balance disorders. Anat Rec, 303:527-543, 2020. © 2019 American Association for Anatomy.

P2RX2 and P2RX4 receptors mediate cation absorption in transitional cells and supporting cells of the utricular macula

Purinergic receptors protect the cochlea during high-intensity stimulation by providing a parallel shunt pathway through non-sensory neighboring epithelial cells for cation absorption. So far, there is no direct functional evidence for the presence and type/subunit of purinergic receptors in the utricle of the vestibular labyrinth. The goal of the present study was to investigate which purinergic receptors are expressed and carry cation-absorption currents in the utricular transitional cells and macula. Purinergic agonists induced cation-absorption currents with a potency order of ATP > bzATP = αβmeATP ≫ ADP = UTP = UDP. ATP and bzATP are full agonists, whereas αβmeATP is a partial agonist. ATP-induced currents were partially inhibited by 100 μM suramin, 10 μM pyridoxal-phosphate-6-azo-(benzene-2,4-disulfonic acid (PPADS), or 5 μM 5-(3-bromophenyl)-1,3-dihydro-2H-benzofuro[3,2-e]-1, 4-diazepin-2-one (5-BDBD), and almost completely blocked by 100 μM Gd3+ or by a combination of 10 μM PPADS and 5 μM 5-BDBD. Expression of the P2RX2 and P2RX4 receptor was detected by immunocytochemistry in transitional cells and macular supporting cells. This is the first study to demonstrate that ATP induces cation currents carried by a combination of P2RX2 and P2RX4 in utricular transitional and macular epithelial cells, and supporting the hypothesis that purinergic receptors protect utricular hair cells during elevated stimulus intensity levels.

Acoustic ocular vestibulat evoked miogenic ptotentials (AC-oVEMP) in assessment of vertigo

Evaluation of acoustic vestibular evoked myogenic potentials (AC-VEMPs) is one of the tests performed to assess the function of the balance system. Evaluation of acoustic ocular vestibular evoked myogenic potentials (AC-oVEMPs) consists in the measurement of potentials evoked within oculomotor muscles, particularly the inferior oblique muscles (the most superficial muscles) with impulses being transmitted along the superior branch of the vestibular nerve from the utricular macula. Despite the fact that the measurement of oVEMPs was introduced relatively recently, it has found widespread use as a diagnostic tool in otology and neurotology. Despite the growing number of literature reports, the methodology of the test and the methods for the interpretation of its results are still subject to debate. This article is dedicated to various aspects of AC-oVEMP tests, including the methodology of the test, interpretation of its results and potential use in the diagnostics of vertigo.

Ultrastructural Characterization of Stem Cell-Derived Replacement Vestibular Hair Cells Within Ototoxin-Damaged Rat Utricle Explants.

ABSTRACTThe auditory apparatus of the inner ear does not show turnover of sensory hair cells (HCs) in adult mammals; in contrast, there are many observations supporting low‐level turnover of vestibular HCs within the balance organs of mammalian inner ears. This low‐level renewal of vestibular HCs exists during normal conditions and it is further enhanced after trauma‐induced loss of these HCs. The main process for renewal of HCs within mammalian vestibular epithelia is a conversion/transdifferentiation of existing supporting cells (SCs) into replacement HCs.In earlier studies using long‐term organ cultures of postnatal rat macula utriculi, HC loss induced by gentamicin resulted in an initial substantial decline in HC density followed by a significant increase in the proportion of HCs to SCs indicating the production of replacement HCs. In the present study, using the same model of ototoxic damage to study renewal of vestibular HCs, we focus on the ultrastructural characteristics of SCs undergoing transdifferentiation into new HCs. Our objective was to search for morphological signs of SC plasticity during this process. In the utricular epithelia, we observed immature HCs, which appear to be SCs transdifferentiating into HCs. These bridge SCs have unique morphological features characterized by formation of foot processes, basal accumulation of mitochondria, and an increased amount of connections with nearby SCs. No gap junctions were observed on these transitional cells. The tight junction seals were morphologically intact in both control and gentamicin‐exposed explants. Anat Rec, 303:506–515, 2020. © 2019 The Authors. The Anatomical Record published by Wiley Periodicals, Inc. on behalf of American Association of Anatomists.

Peripheral Vestibular Organ Degeneration After Temporal Bone Fracture: A Human Otopathology Study.

Vestibular symptoms are a common sequela of temporal bone fractures (TBFs). The mechanisms of injury to the peripheral vestibular system following TBF, however, are not well described. Herein, we aimed to investigate the histopathology of the peripheral vestibular system in patients who sustained TBFs. Retrospective human specimen analysis. Specimens from the National Temporal Bone Pathology Registry with (cases) and without (controls) TBFs were evaluated. Specimens were analyzed by light microscopy for vestibular hair cell and/or dendritic degeneration, presence of endolymphatic hydrops, blockage of the endolymphatic duct, and number of Scarpa ganglion cells (ScGCs) in the superior and inferior vestibular nerves. Seven temporal bones (TBs) from five individuals with TBFs, and seven TBs from six age-matched individuals without a history of head injury met inclusion and exclusion criteria. All fractures involved the otic capsule. Severe degeneration of the cristae was identified in the semicircular canals in all TBF cases. The utricular and saccular maculae showed mild to severe degeneration in the TBF cases. Vestibular hydrops (n = 2 TBs) and blockage of the endolymphatic duct (n = 3 TBs) were also present in the TBF cases. There was a decrease of 52.6% in the mean total ScGC count in the TBF cases (n = 3 TBs) compared to age-matched controls (n = 7 TBs, P = .015). There was a mean loss of 53% of the ScGCs in the superior vestibular nerve and a mean loss of 52.3% of the ScGCs in the inferior vestibular nerve compared to age-matched controls (P = .033 and P = .021, respectively). In a cohort of patients with TBFs, there were distinct peripheral vestibular changes including reduction of ScGCs. NA Laryngoscope, 130:752-760, 2020.

Spontaneous and Acetylcholine Evoked Calcium Transients in the Developing Mouse Utricle.

Spontaneous calcium transients are present during early postnatal development in the mouse retina and cochlea, and play an important role in maturation of the sensory organs and neural circuits in the central nervous system (CNS). It is not known whether similar calcium transients occur during postnatal development in the vestibular sensory organs. Here we demonstrate spontaneous intracellular calcium transients in sensory hair cells (HCs) and supporting cells (SCs) in the murine utricular macula during the first two postnatal weeks. Calcium transients were monitored using a genetically encoded calcium indicator, GCaMP5G (G5), at 100 ms-frame−1 in excised utricle sensory epithelia, including HCs, SCs, and neurons. The reporter line expressed G5 and tdTomato (tdT) in a Gad2-Cre dependent manner within a subset of utricular HCs, SCs and neurons. Kinetics of the G5 reporter limited temporal resolution to calcium events lasting longer than 200 ms. Spontaneous calcium transients lasting 1-2 s were observed in the expressing population of HCs at birth and slower spontaneous transients lasting 10-30 s appeared in SCs by P3. Beginning at P5, calcium transients could be modulated by application of the efferent neurotransmitter acetylcholine (ACh). In mature mice, calcium transients in the utricular macula occurred spontaneously, had a duration 1-2 s, and could be modulated by the exogenous application of acetylcholine (ACh) or muscarine. Long-lasting calcium transients evoked by ACh in mature mice were blocked by atropine, consistent with previous reports describing the role of muscarinic receptors expressed in calyx bearing afferents in efferent control of vestibular sensation. Large spontaneous and ACh evoked transients were reversibly blocked by the inositol trisphosphate receptor (IP3R) antagonist aminoethoxydiphenyl borate (2-APB). Results demonstrate long-lasting calcium transients are present in the utricular macula during the first postnatal week, and that responses to ACh mature over this same time period.

Utricular function in vestibular neuritis: a pilot study of concordance/discordance between ocular vestibular evoked myogenic potentials and ocular cycloposition.

Vestibular neuritis (VN) can affect utricular afferents. Utricular function can be assessed by ocular vestibular evoked myogenic potentials (oVEMPs) whose abnormalities include weak or absent responses, and ocular cycloposition whose abnormalities include ocular torsion (OT). When studied independently in vestibular neuritis, oVEMPs are abnormal in 61-82% of cases, and OT is present in 72-80% of cases. The similar range of abnormalities suggests the hypothesis that these tests should be concordantly abnormal. We tested this hypothesis by identifying consecutive adult cases of VN in whom both oVEMPs and OT were performed. OT and oVEMP overlapped (both were abnormal) in only 47% of cases. In 40% of cases oVEMPs alone were abnormal, and in 13% of cases, OT alone was present. These results suggest that oVEMPs and OT assess different aspects of utricular function believed to arise from discrete zones of the utricular macula; the former are thought to reflect the activity of extra-striolar afferents (which detect constant acceleration), and the latter reflects the activity of striolar afferents (which detect change in acceleration).

Investigation of inner ear anatomy in mouse using X-ray phase contrast tomography.

A thorough understanding of inner ear anatomy is important for investigators. However, investigation of the mouse inner ear is difficult due to the limitations of imaging techniques. X-ray phase contrast tomography increases contrast 100-1,000 times compared with conventional X-ray imaging. This study aimed to investigate inner ear anatomy in a fresh post-mortem mouse using X-ray phase contrast tomography and to provide a comprehensive atlas of microstructures with less tissue deformation. All experiments were performed in accordance with our institution's guidelines on the care and use of laboratory animals. A fresh mouse cadaver was scanned immediately after sacrifice using an inline phase contrast tomography system. Slice images were reconstructed using a filtered back-projection (FBP) algorithm. Standardized axial and coronal planes were adjusted with a multi-planar reconstruction method. Some three-dimensional (3D) objects were reconstructed by surface rendering. The characteristic features of microstructures, including otoconia masses of the saccular and utricular maculae, superior and inferior macula cribrosae, single canal, modiolus, and osseous spiral lamina, were described in detail. Spatial positions and relationships of the vestibular structures were exhibited in 3D views. This study investigated mouse inner ear anatomy and provided a standardized presentation of microstructures. In particular, otoconia masses were visualized in their natural status without contrast for the first time. The comprehensive anatomy atlas presented in this study provides an excellent reference for morphology studies of the inner ear.

A New Model for Congenital Vestibular Disorders.

Many developmental disorders of the inner ear are manifested clinically as delayed motor development and challenges in maintaining posture and balance, indicating involvement of central vestibular circuits. How the vestibular circuitry is rewired in pediatric cases is poorly understood due to lack of a suitable animal model. Based on this, our lab designed and validated a chick embryo model to study vestibular development in congenital vestibular disorders. The developing inner ear or "otocyst" on the right side of 2-day-old chick embryos (E2) was surgically rotated 180° in the anterior-posterior axis, forming the "anterior-posterior axis rotated otocyst chick" or ARO chick. The ARO chick has a reproducible pathology of a sac with truncated or missing semicircular canals. A sac is the most common inner ear defect found in children with congenital vestibular disorders. In E13 ARO chicks, the sac contained all three cristae and maculae utriculi and sacculi, but the superior crista and macula utriculi were shortened in anterior-posterior extent. Also, the number of principal cells of the tangential vestibular nucleus, a major avian vestibular nucleus, was decreased 66% on the rotated side. After hatching, no difference was detected between ARO and normal chicks in their righting reflex times. However, unlike normal chicks, ARO hatchlings had a constant, right head tilt, and after performing the righting reflex, ARO chicks stumbled and walked with a widened base. Identifying the structure and function of abnormally developed brain regions in ARO chicks may assist in improving treatments for patients with congenital vestibular disorder.

Peripheral Vestibular System Histopathologic Changes following Head Injury without Temporal Bone Fracture.

Vestibular symptoms such as dizziness and vertigo are common after head injury and may be due to trauma to the peripheral vestibular system. The pathophysiology of peripheral vestibular symptoms following head injury without temporal bone (TB) fracture, however, is not well understood. Herein, we investigate the histopathology of the peripheral vestibular system of patients who sustained head injury without a TB fracture. Otopathology study. Otopathology laboratory. TB of subjects with a history of head injury without TB fractures were included and evaluated by light microscopy. Specimens were assessed for qualitative and quantitative characteristics, such as number of Scarpa's ganglion cells in the superior and inferior vestibular nerves, vestibular hair cell and/or dendrite degeneration in vestibular end organs, presence of vestibular hydrops, and obstruction of the endolymphatic duct. Five cases (n = 5 TBs) had evidence of vestibular pathology. There was a decrease of 48.6% (range, 40%-59%) in the mean count of Scarpa's ganglion cells as compared with that of normative historical age-matched controls. Moderate to severe degeneration of the vestibular membranous labyrinth was identified in the posterior, superior, and lateral canals in several cases (50%, n = 4 TBs). The maculae utriculi and sacculi showed mild to severe degeneration in 2 cases. Additional findings include vestibular hydrops (25%, n = 2 TBs) and blockage of the endolymphatic duct (n = 1 TB). Otopathologic analysis of patients with a history of head injury without TB fracture demonstrated peripheral vestibular otopathology. Future studies are necessary to determine if otopathology findings are directly attributable to head injury.

Oxidative Stress in the Blood Labyrinthine Barrier in the Macula Utricle of Meniere's Disease Patients.

The blood labyrinthine barrier (BLB) is critical in the maintenance of inner ear ionic and fluid homeostasis. Recent studies using imaging and histopathology demonstrate loss of integrity of the BLB in the affected inner ear of Meniere’s disease (MD) patients. We hypothesized that oxidative stress is involved in the pathogenesis of BLB degeneration, and to date there are no studies of oxidative stress proteins in the human BLB. We investigated the ultrastructural and immunohistochemical changes of the BLB in the vestibular endorgan, the macula utricle, from patients with MD (n = 10), acoustic neuroma (AN) (n = 6) and normative autopsy specimens (n = 3) with no inner ear disease. Each subject had a well-documented clinical history and audiovestibular testing. Utricular maculae were studied using light and transmission electron microscopy and double labeling immunofluorescence. Vascular endothelial cells (VECs) were identified using isolectin B4 (IB4) and glucose-transporter-1 (GLUT-1). Pericytes were identified using alpha smooth muscle actin (αSMA) and phalloidin. IB4 staining of VECS was consistently seen in both AN and normative. In contrast, IB4 was nearly undetectable in all MD specimens, consistent with the significant VEC damage confirmed on transmission electron microscopy. GLUT-1 was present in MD, AN, and normative. αSMA and phalloidin were expressed consistently in the BLB pericytes in normative, AN specimen, and Meniere’s specimens. Endothelial nitric oxide synthase (eNOS), inducible nitric oxide synthase (iNOS), and nitrotyrosine were used as markers of oxidative stress. The VECs of the BLB in Meniere’s had significantly higher levels of expression of iNOS and nitrotyrosine compared with normative and AN specimen. eNOS-IF staining showed similar patterns in normative and Meniere’s specimens. Microarray-based gene expression profiling confirmed upregulation of iNOS mRNA from the macula utricle of Meniere’s patients compared with AN. Nitrotyrosine, a marker recognized as a hallmark of inflammation, especially when seen in association with an upregulation of iNOS, was detected in the epithelial and stromal cells in addition to VECs in MD. Immunohistochemical and ultrastructural degenerative changes of the VEC suggest that these cells are the primary targets of oxidative stress, and pericyte pathology including degeneration and migration, likely also plays a role in the loss of integrity of the BLB and triggering of inflammatory pathways in MD. These studies advance our scientific understanding of oxidative stress in the human inner ear BLB and otopathology.