Vestibular dysfunction in the adult CBA/CaJ mouse after lead and cadmium treatment.

OBJECTIVES: The vestibular system allows the perception of position and motion and its dysfunction presents as motion impairment, vertigo and balance abnormalities, leading to debilitating psychological discomfort and difficulty performing daily tasks. Although declines and deficits in vestibular function have been noted in rats exposed to lead (Pb) and in humans exposed to Pb and cadmium (Cd), no studies have directly examined the pathological and pathophysiological effects upon the vestibular apparatus of the inner ear. METHODS: Eighteen young adult mice were exposed through their drinking water (3 mM Pb, 300 µM Cd, or a control treatment) for 10 weeks. Before and after treatment, they underwent a vestibular assessment, consisting of a rotarod performance test and a novel head stability test to measure the vestibulocolic reflex. At the conclusion of the study, the utricles were analyzed immunohistologically for condition of hair cells and nerve fibers. RESULTS: Increased levels of Pb exposure correlated with decreased head stability in space; no significant decline in performance on rotarod test was found. No damage to the hair cells or the nerve fibers of the utricle was observed in histology. CONCLUSIONS: The young adult CBA/CaJ mouse is able to tolerate occupationally-relevant Pb and Cd exposure well, but the correlation between Pb exposure and reduced head stability suggests that Pb exposure causes a decline in vestibular function. © 2016 Wiley Periodicals, Inc. Environ Toxicol, 2016. © 2016 Wiley Periodicals, Inc. Environ Toxicol 32: 869-876, 2017.

Photoresponse diversity among the five types of intrinsically photosensitive retinal ganglion cells.

Intrinsically photosensitive retinal ganglion cells (ipRGCs) mediate non-image-forming visual responses, including pupillary constriction, circadian photoentrainment and suppression of pineal melatonin secretion. Five morphological types of ipRGCs, M1-M5, have been identified in mice. In order to understand their functions better, we studied the photoresponses of all five cell types, by whole-cell recording from fluorescently labelled ipRGCs visualized using multiphoton microscopy. All ipRGC types generated melanopsin-based ('intrinsic') as well as synaptically driven ('extrinsic') light responses. The intrinsic photoresponses of M1 cells were lower threshold, higher amplitude and faster than those of M2-M5. The peak amplitudes of extrinsic light responses differed among the ipRGC types; however, the responses of all cell types had comparable thresholds, kinetics and waveforms, and all cells received rod input. While all five types exhibited inhibitory amacrine-cell and excitatory bipolar-cell inputs from the 'on' channel, M1 and M3 received additional 'off'-channel inhibition, possibly through their 'off'-sublamina dendrites. The M2-M5 ipRGCs had centre-surround-organized receptive fields, implicating a capacity to detect spatial contrast. In contrast, the receptive fields of M1 cells lacked surround antagonism, which might be caused by the surround of the inhibitory input nullifying the surround of the excitatory input. All ipRGCs responded robustly to a wide range of motion speeds, and M1-M4 cells appeared tuned to different speeds, suggesting that they might analyse the speed of motion. Retrograde labelling revealed that M1-M4 cells project to the superior colliculus, suggesting that the contrast and motion information signalled by these cells could be used by this sensorimotor area to detect novel objects and motion in the visual field.

Wavelet transform of single-trial vestibular short-latency evoked potential reveals temporary reduction in signal detectability and temporal precision following noise exposure.

The vestibular short-latency evoked potential (VsEP) reflects the activity of irregular vestibular afferents and their target neurons in the brain stem. Attenuation of trial-averaged VsEP waveforms is widely accepted as an indicator of vestibular dysfunction, however, more quantitative analyses of VsEP waveforms could reveal underlying neural properties of VsEP waveforms. Here, we present a time-frequency analysis of the VsEP with a wavelet transform on a single-trial basis, which allows us to examine trial-by-trial variability in the strength of VsEP waves as well as their temporal coherence across trials. Using this method, we examined changes in the VsEP following 110 dB SPL noise exposure in rats. We found detectability of head jerks based on the power of wavelet transform coefficients was significantly reduced 1 day after noise exposure but recovered nearly to pre-exposure level in 3 - 7 days and completely by 28 days after exposure. Temporal coherence of VsEP waves across trials was also significantly reduced on 1 day after exposure but recovered with a similar time course. Additionally, we found a significant reduction in the number of calretinin-positive calyces in the sacculi collected 28 days after noise exposure. Furthermore, the number of calretinin-positive calyces was significantly correlated with the degree of reduction in temporal coherence and/or signal detectability of the smallest-amplitude jerks. This new analysis of the VsEP provides more quantitative descriptions of noise-induced changes as well as new insights into potential mechanisms underlying noise-induced vestibular dysfunction. Significance Statement: Our study presents a new method of VsEP quantification using wavelet transform on a single-trial basis. It also describes a novel approach to determine the stimulus threshold of the VsEP based on signal-detection theory and Rayleigh statistics. The present analysis could also be applied to analysis of auditory brain stem response (ABR). Thus, it has the potential to provide new insights into the physiological properties that underlie peripheral vestibular and auditory dysfunction.

Balance beam crossing times are slower after noise exposure in rats.

Introduction: The vestibular system integrates signals related to vision, head position, gravity, motion, and body position to provide stability during motion through the environment. Disruption in any of these systems can reduce agility and lead to changes in ability to safely navigate one's environment. Causes of vestibular decline are diverse; however, excessive noise exposure can lead to otolith organ dysfunction. Specifically, 120 decibel (dB) sound pressure level (SPL) 1.5 kHz-centered 3-octave band noise (1.5 kHz 3OBN) causes peripheral vestibular dysfunction in rats, measured by vestibular short-latency evoked potential (VsEP) and reduced calretinin-immunolabeling of calyx-only afferent terminals in the striolar region of the saccule. The present study examined the functional impact of this noise exposure condition, examining changes in motor performance after noise exposure with a balance beam crossing task. Methods: Balance beam crossing time in rats was assessed for 19 weeks before and 5 weeks after noise exposure. Balance beam crossings were scored to assess proficiency in the task. When animals were proficient, they received a single exposure to 120 dB SPL 3-octave band noise. Results: During the initial training phase slower crossing times and higher scores, including multiple failures were observed. This was followed by a period of significant improvement leading to proficiency, characterized by fast and stable crossing times and consistently low scores. After noise exposure, crossing times were significantly elevated from baseline for 4-weeks. A total of 5 weeks after noise exposure, crossing times improved, and though still trending higher than baseline, they were no longer significantly different from baseline. Discussion: These findings show that the noise-induced peripheral vestibular changes we previously observed at cellular and electro-physiological levels also have an impact at a functional level. It has been previously shown that imbalance is associated with slower walking speed in older adults and aged rats. These findings in noise-exposed rats may have implications for people who experience noisy environments and for seniors with a history of noise exposure who also experience balance disorders and may be at increased fall risk.

Exposure to Intense Noise Causes Vestibular Loss.

INTRODUCTION: The vestibular system is essential for normal postural control and balance. Because of their proximity to the cochlea, the otolith organs are vulnerable to noise. We previously showed that head jerks that evoke vestibular nerve activity were no longer capable of inducing a response after noise overstimulation. The present study adds a greater range of jerk intensities to determine if the response was abolished or required more intense stimulation (threshold shift). MATERIALS AND METHODS: Vestibular short-latency evoked potential (VsEP) measurements were taken before noise exposure and compared to repeated measurements taken at specific time points for 28 days after noise exposure. Calretinin was used to identify changes in calyx-only afferents in the sacculus. RESULTS: Results showed that more intense jerk stimuli could generate a VsEP, although it was severely attenuated relative to prenoise values. When the VsEP was evaluated 4 weeks after noise exposure, partial recovery was observed. CONCLUSION: These data suggest that noise overstimulation, such as can occur in the military, could introduce an increased risk of imbalance that should be evaluated before returning a subject to situations that require normal agility and motion. Moreover, although there is recovery with time, some dysfunction persists for extended periods.

Effects of Noise Exposure on the Vestibular System: A Systematic Review.

Despite our understanding of the impact of noise-induced damage to the auditory system, much less is known about the impact of noise exposure on the vestibular system. In this article, we review the anatomical, physiological, and functional evidence for noise-induced damage to peripheral and central vestibular structures. Morphological studies in several animal models have demonstrated cellular damage throughout the peripheral vestibular system and particularly in the otolith organs; however, there is a paucity of data on the effect of noise exposure on human vestibular end organs. Physiological studies have corroborated morphological studies by demonstrating disruption across vestibular pathways with otolith-mediated pathways impacted more than semicircular canal-mediated pathways. Similar to the temporary threshold shifts observed in the auditory system, physiological studies in animals have suggested a capacity for recovery following noise-induced vestibular damage. Human studies have demonstrated that diminished sacculo-collic responses are related to the severity of noise-induced hearing loss, and dose-dependent vestibular deficits following noise exposure have been corroborated in animal models. Further work is needed to better understand the physiological and functional consequences of noise-induced vestibular impairment in animals and humans.

The Interaction of Pre-programmed Eye Movements With the Vestibulo-Ocular Reflex.

The Vestibulo-Ocular Reflex (VOR) works to stabilize gaze during unexpected head movements. However, even subjects who lack a VOR (e.g., vestibulopathic patients) can achieve gaze stability during planned head movements by using pre-programmed eye movements (PPEM). The extent to which PPEM are used by healthy intact subjects and how they interact with the VOR is still unclear. We propose a model of gaze stabilization which makes several claims: (1) the VOR provides ocular stability during unexpected (i.e., passive) head movements; (2) PPEM are used by both healthy and vestibulopathic subjects during planned (i.e., active) head movements; and (3) when a passive perturbation interrupts an active head movement in intact animals (i.e., combined passive and active head movement) the VOR works with PPEM to provide compensation. First, we show how our model can reconcile some seemingly conflicting findings in earlier literature. We then test the above-mentioned predictions against data we collected from both healthy and vestibular-lesioned guinea pigs. We found that (1) vestibular-lesioned animals showed a dramatic decrease in compensatory eye movements during passive head movements, (2) both populations showed improved ocular compensation during active vs. passive head movements, and (3) during combined active and passive head movements, eye movements compensated for both the active and passive component of head velocity. These results support our hypothesis that while the VOR provides compensation during passive head movements, PPEM are used by both intact and lesioned subjects during active movements and further, that PPEM work together with the VOR to achieve gaze stability.

Mice with conditional deletion of Cx26 exhibit no vestibular phenotype despite secondary loss of Cx30 in the vestibular end organs.

Connexins are components of gap junctions which facilitate transfer of small molecules between cells. One member of the connexin family, Connexin 26 (Cx26), is prevalent in gap junctions in sensory epithelia of the inner ear. Mutations of GJB2, the gene encoding Cx26, cause significant hearing loss in humans. The vestibular system, however, does not usually show significant functional deficits in humans with this mutation. Mouse models for loss of Cx26 function demonstrate hearing loss and cochlear pathology but the extent of vestibular dysfunction and organ pathology are less well characterized. To understand the vestibular effects of Cx26 mutations, we evaluated vestibular function and histology of the vestibular sensory epithelia in a conditional knockout (CKO) mouse with Cx26 loss of function. Transgenic C57BL/6 mice, in which cre-Sox10 drives excision of the Cx26 gene from non-sensory cells flanking the sensory epithelium of the inner ear (Gjb2-CKO), were compared to age-matched wild types. Animals were sacrificed at ages between 4 and 40 weeks and their cochlear and vestibular sensory organs harvested for histological examination. Cx26 immunoreactivity was prominent in the peripheral vestibular system and the cochlea of wild type mice, but absent in the Gjb2-CKO specimens. The hair cell population in the cochleae of the Gjb2-CKO mice was severely depleted but in the vestibular organs it was intact, despite absence of Cx26 expression. The vestibular organs appeared normal at the latest time point examined, 40 weeks. To determine whether compensation by another connexin explains survival of the normal vestibular sensory epithelium, we evaluated the presence of Cx30 in the Gjb2-CKO mouse. We found that Cx30 labeling was normal in the cochlea, but it was decreased or absent in the vestibular system. The vestibular phenotype of the mutants was not different from wild-types as determined by time on the rotarod, head stability tests and physiological responses to vestibular stimulation. Thus presence of Cx30 in the cochlea does not compensate for Cx26 loss, and the absence of both connexins from vestibular sensory epithelia is no more injurious than the absence of one of them. Further studies to uncover the physiological foundation for this difference between the cochlea and the vestibular organs may help in designing treatments for GJB2 mutations.

Hippocampal long-term potentiation (LTP) is reduced by a coplanar PCB congener.

Neurotoxicity of polychlorinated biphenyls (PCBs) is usually ascribed to the ortho-substituted congeners. We have examined the effects of acute perfusion of 3,3',4,4'-tetrachlorobiphenyl (PCB 77), a coplanar, dioxin-like congener, on long-term potentiation (LTP) in the Schaffer collateral-CA1 and the mossy fiber-CA3 pathways in mouse hippocampus. LTP in both pathways was blocked by PCB 77, with a threshold effect at a concentration of 1 microM. LTP is a useful model of learning and memory function in which a patterned stimulation of an afferent pathway produces a persistent increase in the efficacy of synaptic transmission. LTP is reduced by PCB mixtures and ortho-substituted congeners at concentrations comparable to those studied here. These observations provide evidence in support of the hypothesis that dioxin-like and non-dioxin-like PCB congeners are equally potent in causing the cognitive decrements seen in children exposed prenatally to PCBs.

Association between hearing loss and saccular dysfunction in older individuals.

OBJECTIVE: 1) Describe the association between hearing loss and dysfunction of each of the 5 vestibular end-organs--the horizontal, superior, and posterior semicircular canals; saccule; and utricle--in older individuals. 2) Evaluate whether hearing loss and vestibular end-organ deficits share any risk factors. STUDY DESIGN: Cross-sectional study. SETTING: Academic medical center. PATIENTS: Fifty-one individuals age 70 years or older. INTERVENTIONS: Audiometry, head-thrust dynamic visual acuity (htDVA), sound-evoked cervical vestibular-evoked myogenic potential (cVEMP), and tap-evoked ocular VEMP (oVEMP). MAIN OUTCOME MEASURES: Audiometric pure-tone averages (PTA), htDVA LogMAR scores as a measure of semicircular canal function in each canal plane, and cVEMP and oVEMP amplitudes as a measure of saccular and utricular function, respectively. RESULTS: We observed a significant correlation between hearing loss at high frequencies and reduced cVEMP amplitudes (or reduced saccular function; r = -0.37, p < 0.0001) in subjects age 70 years or older. In contrast, hearing loss was not associated with oVEMP amplitudes (or utricular function), or htDVA LogMAR scores (or semicircular canal function) in any of the canal planes. Age and noise exposure were significantly associated with measures of both cochlear and saccular dysfunction. CONCLUSION: The concomitant decline in the cochlear and saccular function associated with aging may reflect their common embryologic origin in the pars inferior of the labyrinth. Noise exposure seems to be related to both saccular and cochlear dysfunction. These findings suggest a potential benefit of screening individuals with presbycusis-particularly those with significant noise exposure history-for saccular dysfunction, which may contribute to fall risk in the elderly.

Tests of models for saccade-vergence interaction using novel stimulus conditions.

During natural activities, two types of eye movements - saccades and vergence - are used in concert to point the fovea of each eye at features of interest. Some electrophysiological studies support the concept of independent neurobiological substrates for saccades and vergence, namely saccadic and vergence burst neurons. Discerning the interaction of these two components is complicated by the near-synchronous occurrence of saccadic and vergence components. However, by positioning the far target below the near target, it is possible to induce responses in which the peak velocity of the vertical saccadic component precedes the peak velocity of the horizontal vergence component by approximately 75 ms. When saccade-vergence responses are temporally dissociated in this way, the vergence velocity waveform changes, becoming less skewed. We excluded the possibility that such change in skewing was due to visual feedback by showing that similar behavior occurred in darkness. We then tested a saccade-related vergence burst neuron (SVBN) model proposed by Zee et al. in J Neurophysiol 68:1624-1641 (1992), in which omnipause neurons remove inhibition from both saccadic and vergence burst neurons. The technique of parameter estimation was used to calculate optimal values for responses from human subjects in which saccadic and convergence components of response were either nearly synchronized or temporally dissociated. Although the SVBN model could account for convergence waveforms when saccadic and vergence components were nearly synchronized, it could not when the components were temporally dissociated. We modified the model so that the saccadic pulse changed the parameter values of the convergence burst units if both components were synchronized. The modified model accounted for velocity waveforms of both synchronous and dissociated convergence movements. We conclude that both the saccadic pulse and omnipause neuron inhibition influence the generation of vergence movements when they are made synchronously with saccades.