The quantal component of synaptic transmission from sensory hair cells to the vestibular calyx.

Spontaneous and stimulus-evoked excitatory postsynaptic currents (EPSCs) were recorded in calyx nerve terminals from the turtle vestibular lagena to quantify key attributes of quantal transmission at this synapse. On average, EPSC events had a magnitude of ∼ 42 pA, a rise time constant of τ(0) ∼ 229 µs, decayed to baseline with a time constant of τ(R) ∼ 690 µs, and carried ∼ 46 fC of charge. Individual EPSCs varied in magnitude and decay time constant. Variability in the EPSC decay time constant was hair cell dependent and due in part to a slow protraction of the EPSC in some cases. Variability in EPSC size was well described by an integer summation of unitary quanta, with each quanta of glutamate gating a unitary postsynaptic current of ∼ 23 pA. The unitary charge was ∼ 26 fC for EPSCs with a simple exponential decay and increased to ∼ 48 fC for EPSCs exhibiting a slow protraction. The EPSC magnitude and the number of simultaneous unitary quanta within each event increased with presynaptic stimulus intensity. During tonic hair cell depolarization, both the EPSC magnitude and event rate exhibited adaptive run down over time. Present data from a reptilian calyx are remarkably similar to noncalyceal vestibular synaptic terminals in diverse species, indicating that the skewed EPSC size distribution and multiquantal release might be an ancestral property of inner ear ribbon synapses.

Evidence that protons act as neurotransmitters at vestibular hair cell-calyx afferent synapses.

Present data support the conclusion that protons serve as an important neurotransmitter to convey excitatory stimuli from inner ear type I vestibular hair cells to postsynaptic calyx nerve terminals. Time-resolved pH imaging revealed stimulus-evoked extrusion of protons from hair cells and a subsequent buildup of [H(+)] within the confined chalice-shaped synaptic cleft (ΔpH ∼ -0.2). Whole-cell voltage-clamp recordings revealed a concomitant nonquantal excitatory postsynaptic current in the calyx terminal that was causally modulated by cleft acidification. The time course of [H(+)] buildup limits the speed of this intercellular signaling mechanism, but for tonic signals such as gravity, protonergic transmission offers a significant metabolic advantage over quantal excitatory postsynaptic currents--an advantage that may have driven the proliferation of postsynaptic calyx terminals in the inner ear vestibular organs of contemporary amniotes.

The anatomical and physiological framework for vestibular prostheses.

This article reviews the structure function of the vestibular system and its pathology with respect to requirements for the design and construction of a functional vestibular prosthesis. The ultimate goal of a vestibular prosthesis is to restore balance and equilibrium through direct activation of vestibular nerve fibers. An overview of the peripheral and central vestibular systems that highlights their most important functional aspects re: the design of a prosthesis is provided. Namely, the peripheral labyrinth faithfully transduces head motion and gravity in both the time and frequency domains. These signals are described in hopes that they may be prosthetically replicated. The peripheral and central connections of the vestibular nerve are also discussed in detail, as are the vestibular nuclei in the brainstem that receive VIIIth nerve innervation. Lastly, the functional effector pathways of the vestibular system, including the vestibulo-ocular, vestibulo-spinal, vestibulo-colic, vestibulo-autonomic, and vestibular efferent innervation of the labyrinth are reviewed.

Infrared photostimulation of the crista ampullaris.

The present results show that the semicircular canal crista ampullaris of the toadfish, Opsanus tau, is sensitive to infrared radiation (IR) applied in vivo. IR pulse trains (∼1862 nm, ∼200 µs pulse⁻¹) delivered to the sensory epithelium by an optical fibre evoked profound changes in phasic and tonic discharge rates of postsynaptic afferent neurons. Phasic afferent responses to pulsed IR occurred with a latency of <8 ms while tonic responses developed with a time constant (τ) of 7 ms to 10 s following the onset or cessation of the radiation. Afferents responded to direct optical radiation of the sensory epithelium but did not respond to thermal stimuli that generated nearly equivalent temperature increases of the whole organ. A subset of afferent neurons fired an action potential in response to each IR pulse delivered to the sensory epithelium, at phase-locked rates up to 96 pulses per second. The latency between IR pulses and afferent nerve action potentials was much greater than synaptic delay and spike generation, demonstrating the presence of a signalling delay interposed between the IR pulse and the action potential. The same IR stimulus applied to afferent nerve axons failed to evoke responses of similar magnitude and failed to phase-lock afferent nerve action potentials. The present data support the hypothesis that pulsed IR activates sensory hair cells, thus leading to modulation of synaptic transmission and afferent nerve discharge reported here.

Golgi cells operate as state-specific temporal filters at the input stage of the cerebellar cortex.

Cerebellar processing of incoming information begins at the synapse between mossy fibers and granule cells, a synapse that is strongly controlled through Golgi cell inhibition. Thus, Golgi cells are uniquely positioned to control the flow of information into the cerebellar cortex and understanding their responses during behavior is essential to understanding cerebellar function. Here we show, for the first time, that Golgi cells express a unique oculomotor-related signal that can be used to provide state- and time-specific filtering of granule cell activity. We used newly established criteria to identify the unique electrophysiological signature of Golgi cells and recorded these neurons in the squirrel monkey ventral paraflocculus during oculomotor behaviors. We found that they carry eye movement, but not vestibular or visual, information and that this eye movement information is only expressed within a specific range of eye positions for each neuron. In addition, simultaneous recordings of Golgi cells and nearby mossy fibers revealed that Golgi cells have the opposite directional tuning of the mossy fiber(s) that likely drive their responses, and that these responses are more sluggish than their mossy fiber counterparts. Because the mossy fiber inputs appear to convey the activity of burst-tonic neurons in the brainstem, Golgi cell responses reflect a time-filtered negative image of the motor command sent to the extraocular muscles. We suggest a role for Golgi cells in the construction of forward models of movement, commonly hypothesized as a major function of the cerebellar cortex in motor control.

Mechanical amplification by hair cells in the semicircular canals.

Sensory hair cells are the essential mechanotransducers of the inner ear, responsible not only for the transduction of sound and motion stimuli but also, remarkably, for nanomechanical amplification of sensory stimuli. Here we show that semicircular canal hair cells generate a mechanical nonlinearity in vivo that increases sensitivity to angular motion by amplification at low stimulus strengths. Sensitivity at high stimulus strengths is linear and shows no evidence of amplification. Results suggest that the mechanical work done by hair cells contributes approximately 97 zJ/cell of amplification per stimulus cycle, improving sensitivity to angular velocity stimuli below approximately 5 degrees /s (0.3-Hz sinusoidal motion). We further show that mechanical amplification can be inhibited by the brain via activation of efferent synaptic contacts on hair cells. The experimental model was the oyster toadfish, Opsanus tau. Physiological manifestation of mechanical amplification and efferent control in a teleost vestibular organ suggests the active motor process in sensory hair cells is ancestral. The biophysical basis of the motor(s) remains hypothetical, but a key discriminating question may involve how changes in somatic electrical impedance evoked by efferent synaptic action alter function of the motor(s).

Efferent control of hair cell and afferent responses in the semicircular canals.

The sensations of sound and motion generated by the inner ear are controlled by the brain through extensive centripetal innervation originating within the brain stem. In the semicircular canals, brain stem efferent neurons make synaptic contacts with mechanosensory hair cells and with the dendrites of afferent neurons. Here, we examine the relative contributions of efferent action on hair cells and afferents. Experiments were performed in vivo in the oyster toadfish, Opsanus tau. The efferent system was activated via electrical pulses to the brain stem and sensory responses to motion stimuli were quantified by simultaneous voltage recording from afferents and intracellular current- and/or voltage-clamp recordings from hair cells. Results showed synaptic inputs to both afferents and hair cells leading to relatively long-latency intracellular signaling responses: excitatory in afferents and inhibitory in hair cells. Generally, the net effect of efferent action was an increase in afferent background discharge and a simultaneous decrease in gain to angular motion stimuli. Inhibition of hair cells was likely the result of a ligand-gated opening of a major basolateral conductance. The reversal potential of the efferent-evoked current was just below the hair cell resting potential, thus resulting in a small hyperpolarization. The onset latency averaged about 90 ms and latency to peak response was 150-400 ms. Hair cell inhibition often outlasted afferent excitation and, in some cases, latched hair cells in the "off" condition for >1 s following cessation of stimulus. These features endow the animal with a powerful means to adjust the sensitivity and dynamic range of motion sensation.

Dynamic displacement of normal and detached semicircular canal cupula.

The dynamic displacement of the semicircular canal cupula and modulation of afferent nerve discharge were measured simultaneously in response to physiological stimuli in vivo. The adaptation time constant(s) of normal cupulae in response to step stimuli averaged 36 s, corresponding to a mechanical lower corner frequency for sinusoidal stimuli of 0.0044 Hz. For stimuli equivalent to 40-200 deg/s of angular head velocity, the displacement gain of the central region of the cupula averaged 53 nm per deg/s. Afferents adapted more rapidly than the cupula, demonstrating the presence of a relaxation process that contributes significantly to the neural representation of angular head motions by the discharge patterns of canal afferent neurons. We also investigated changes in time constants of the cupula and afferents following detachment of the cupula at its apex-mechanical detachment that occurs in response to excessive transcupular endolymph pressure. Detached cupulae exhibited sharply reduced adaptation time constants (300 ms-3 s, n = 3) and can be explained by endolymph flowing rapidly over the apex of the cupula. Partially detached cupulae reattached and normal afferent discharge patterns were recovered 5-7 h following detachment. This regeneration process may have relevance to the recovery of semicircular canal function following head trauma.

The passive cable properties of hair cell stereocilia and their contribution to somatic capacitance measurements.

Somatic measurements of whole-cell capacitance are routinely used to understand physiologic events occurring in remote portions of cells. These studies often assume the intracellular space is voltage-clamped. We questioned this assumption in auditory and vestibular hair cells with respect to their stereocilia based on earlier studies showing that neurons, with radial dimensions similar to stereocilia, are not always isopotential under voltage-clamp. To explore this, we modeled the stereocilia as passive cables with transduction channels located at their tips. We found that the input capacitance measured at the soma changes when the transduction channels at the tips of the stereocilia are open compared to when the channels are closed. The maximum capacitance is felt with the transducer closed but will decrease as the transducer opens due to a length-dependent voltage drop along the stereocilium length. This potential drop is proportional to the intracellular resistance and stereocilium tip conductance and can produce a maximum capacitance error on the order of fF for single stereocilia and pF for the bundle.

Visual-vestibular interaction in vertical vestibular only neurons.

The central nervous system combines information from different stimulus modalities to generate appropriate behaviors. For instance, vestibular and visual information are combined during oculomotor behavior. We used squirrel monkeys to study this signal combination on vestibular neurons that carry the vertical component of vestibular and visual (slow visual pathway, or optokinetic) signals. We found that these neurons contain a neuronal correlate of asymmetries observed in oculomotor behaviors, and that there is a relationship between their response to vestibular and visual (optokinetic) stimulation. We argue that if this relationship is maintained after learning, changes in one information pathway (e.g. vestibular) will result in changes in the other (e.g. visual), explaining the cross-modality plasticity observed in these systems after vestibulo-ocular reflex motor learning.

Neuronal substrates of motor learning in the velocity storage generated during optokinetic stimulation in the squirrel monkey.

Chronic motor learning in the vestibuloocular reflex (VOR) results in changes in the gain of this reflex and in other eye movements intimately associated with VOR behavior, e.g., the velocity storage generated by optokinetic stimulation (OKN velocity storage). The aim of the present study was to identify the plastic sites responsible for the change in OKN velocity storage after chronic VOR motor learning. We studied the neuronal responses of vertical eye movement flocculus target neurons (FTNs) during the optokinetic after-nystagmus (OKAN) phase of the optokinetic response (OKR) before and after VOR motor learning. Our findings can be summarized as follows. 1) Chronic VOR motor learning changes the horizontal OKN velocity storage in parallel with changes in VOR gain, whereas the vertical OKN velocity storage is more complex, increasing with VOR gain increases, but not changing following VOR gain decreases. 2) FTNs contain an OKAN signal having opposite directional preferences after chronic high versus low gain learning, suggesting a change in the OKN velocity storage representation of FTNs. 3) Changes in the eye-velocity sensitivity of FTNs during OKAN are correlated with changes in the brain stem head-velocity sensitivity of the same neurons. And 4) these changes in eye-velocity sensitivity of FTNs during OKAN support the new behavior after high gain but not low gain learning. Thus we hypothesize that the changes observed in the OKN velocity storage behavior after chronic learning result from changes in brain stem pathways carrying head velocity and OKN velocity storage information, and that a parallel pathway to vertical FTNs changes its OKN velocity storage representation following low, but not high, gain VOR motor learning.

The anatomy of the vestibular nuclei.

The vestibular portion of the eighth cranial nerve informs the brain about the linear and angular movements of the head in space and the position of the head with respect to gravity. The termination sites of these eighth nerve afferents define the territory of the vestibular nuclei in the brainstem. (There is also a subset of afferents that project directly to the cerebellum.) This chapter reviews the anatomical organization of the vestibular nuclei, and the anatomy of the pathways from the nuclei to various target areas in the brain. The cytoarchitectonics of the vestibular brainstem are discussed, since these features have been used to distinguish the individual nuclei. The neurochemical phenotype of vestibular neurons and pathways are also summarized because the chemical anatomy of the system contributes to its signal-processing capabilities. Similarly, the morphologic features of short-axon local circuit neurons and long-axon cells with extrinsic projections are described in detail, since these structural attributes of the neurons are critical to their functional potential. Finally, the composition and hodology of the afferent and efferent pathways of the vestibular nuclei are discussed. In sum, this chapter reviews the morphology, chemoanatomy, connectivity, and synaptology of the vestibular nuclei.

Chronic changes in inputs to dorsal Y neurons accompany VOR motor learning.

Gain changes in the vestibuloocular reflex (VOR) during visual-vestibular mismatch stimulation serve as a model system for motor learning. The cerebellar flocculus and its target neurons in the brain stem (FTN) are candidates for the storage of these novel VOR gains. We have recently studied the changes in vertical flocculus Purkinje cells after chronic VOR motor learning. Recently we recorded Y neurons (a vertical type of FTNs) after chronic VOR motor learning and compared these records with vertical floccular Purkinje cells to document any changes in inputs to FTNs and understand how Y neurons and the vertical Purkinje cells fit into a general model for the vertical VOR. Analysis illustrates that the changes observed in Purkinje cells are not transferred to Y neurons, suggesting that the gain of their synaptic interconnection was modified. We quantified changes in both populations and employed simulations to study changes in parallel pathways to FTNs and to extract the role of the flocculus in VOR adaptation. Low-gain adaptation results in more drastic changes than its high-gain counterpart, causing increases in head velocity sensitivity in parallel pathways. Simulations suggest that cerebellar and brain stem plasticity both participate in novel VOR gain storage and that results obtained following floccular lesion are the product of different mechanisms than those operating in the intact animal.

Report on a workshop concerning the cerebellum and motor learning, held in St Louis October 2004.

A one-day meeting on the cerebellum and motor learning was held in St Louis (October 2004), to address issues arising from a previous larger meeting (Tuebingen, June 2004). The learning tasks considered were VOR adaptation, saccadic adaptation and eyeblink conditioning. A theoretical development was reported that indicated how the cerebellum could use sensory error signals for adaptive control, by decorrelating them from an efferent copy of motor commands. The main topics for discussion were the nature of the error signals actually used by the cerebellum, and the evidence for multiple sites of synaptic plasticity. Reports of studies on VOR adaptation confirmed the presence of error signals in addition to retinal slip, in particular the eye-movement related simple-spike firing of floccular PCs. This firing appears to drive synaptic plasticity in the vestibular nuclei. From a theoretical perspective, a second site of plasticity in the brainstem has two advantages: it improves the high-frequency performance of the VOR given a delayed slip signal, and it allows VOR adaptation when smooth pursuit effectively removes the retinal slip signal. In contrast, some of the physiological data reported on saccadic adaptation seemed incompatible with current theoretical ideas about error signals. However, since other reported data were broadly consistent with those ideas, an important area of experimental disagreement was identified. Furthermore, behavioural studies indicated the presence of multiple sites of plasticity, consistent with earlier lesion studies that suggested one such site within cerebellar cortex and another outside it. Data from eyeblink conditioning suggested that the predictability of the error signal was important. Related ideas have previously emerged from studies of skeletal movement, but their theoretical implications for the cerebellar algorithm have yet to be fully explored. Finally, the long-standing controversy concerning sites of plasticity in eyeblink conditioning illustrated the technical difficulties involved in tracking down such sites.

Determinants of spatial and temporal coding by semicircular canal afferents.

The vestibular semicircular canals are internal sensors that signal the magnitude, direction, and temporal properties of angular head motion. Fluid mechanics within the 3-canal labyrinth code the direction of movement and integrate angular acceleration stimuli over time. Directional coding is accomplished by decomposition of complex angular accelerations into 3 biomechanical components-one component exciting each of the 3 ampullary organs and associated afferent nerve bundles separately. For low-frequency angular motion stimuli, fluid displacement within each canal is proportional to angular acceleration. At higher frequencies, above the lower corner frequency, real-time integration is accomplished by viscous forces arising from the movement of fluid within the slender lumen of each canal. This results in angular velocity sensitive fluid displacements. Reflecting this, a subset of afferent fibers indeed report angular acceleration to the brain for low frequencies of head movement and report angular velocity for higher frequencies. However, a substantial number of afferent fibers also report angular acceleration, or a signal between acceleration and velocity, even at frequencies where the endolymph displacement is known to follow angular head velocity. These non-velocity-sensitive afferent signals cannot be attributed to canal biomechanics alone. The responses of non-velocity-sensitive cells include a mathematical differentiation (first-order or fractional) imparted by hair-cell and/or afferent complexes. This mathematical differentiation from velocity to acceleration cannot be attributed to hair cell ionic currents, but occurs as a result of the dynamics of synaptic transmission between hair cells and their primary afferent fibers. The evidence for this conclusion is reviewed below.

The vestibulo-ocular reflex as a model system for motor learning: what is the role of the cerebellum?

Motor systems are under a continuous adaptive process to maintain behavior throughout developmental changes and disease, a process called motor learning. Simple behaviors with easily measurable inputs and outputs are best suited to understand the neuronal signals that contribute to the required motor learning. Considering simple behaviors, the vestibulo-ocular reflex (VOR) allows quantification of its input and motor output and its neural circuitry is among the best documented. The main candidates for plastic change are the cerebellum and its target neurons in the brainstem. This review focuses on recent data regarding the involvement of the cerebellum in VOR motor learning. Learning can be divided into that acutely acquired over a period of hours and that chronically acquired over longer periods. Both acute and chronic learning have three phases named acquisition, consolidation, and retention. The cerebellar role in retention is disputed, but there is a consensus on the need of an intact cerebellum for acquisition. Data from neuronal recording, lesion studies and transgenic mouse experiments is complex but suggests that the signal representation in the cerebellum contains aspects of both motor output and sensory input. The cerebellum apparently uses different mechanisms for acute and chronic learning as well as for increases and decreases in VOR gain. Recent studies also suggest that the signal content in the cerebellum changes following learning and that the mechanisms used for chronic adaptation involve not only changes in a head velocity component but also in the efference copy of an eye movement command signal reaching Purkinje cells. This data leads to a new conceptual framework having implications for developing theories on the role of the cerebellum in motor learning and in the search for plastic elements within the VOR circuitry. For chronic learning we hypothesize that changes in the head velocity information traveling through the circuitry occur in parallel with changes in the integrator pathway and the efference copy pathway. We further propose that these changes are necessary to maintain the broadband characteristics of the learned behavior.

Convergence of excitatory and inhibitory hair cell transmitters shapes vestibular afferent responses.

The vestibular semicircular canals respond to angular acceleration that is integrated to angular velocity by the biofluid mechanics of the canals and is the primary origin of afferent responses encoding velocity. Surprisingly, some afferents actually report angular acceleration. Our data indicate that hair-cell/afferent synapses introduce a mathematical derivative in these afferents that partially cancels the biomechanical integration and results in discharge rates encoding angular acceleration. We examined the role of convergent synaptic inputs from hair cells to this mathematical differentiation. A significant reduction in the order of the differentiation was observed for low-frequency stimuli after gamma-aminobutyric acid type B receptor antagonist administration. Results demonstrate that gamma-aminobutyric acid participates in shaping the temporal dynamics of afferent responses.

Gamma-aminobutyric acid is present in a spatially discrete subpopulation of hair cells in the crista ampullaris of the toadfish Opsanus tau.

Although gamma-aminobutyric acid (GABA) and glutamate are known to be present in the vestibular sensory epithelia of a variety of species, the functional relationship between these two transmitters is not clear. The present study addresses the three-dimensional spatial distribution of GABA and glutamate immunoreactivity in the vestibular labyrinth of the oyster toadfish by using whole end organs labeled by immunofluorescence with monoclonal anti-GABA and/or antiglutamate antibodies and visualized as whole mounts by multiphoton confocal microscopy. We find glutamate-immunoreactive hair cells present throughout the sensory epithelium. In contrast, prominent GABA immunoreactivity is restricted to a small population of hair cells located in the central region of the crista. Double immunofluorescence reveals two distinct staining patterns in GABA-labeled hair cells. Most ( approximately 80%) GABA-labeled cells show trace levels of glutamate, appropriate for the metabolic/synthetic role of cytoplasmic glutamate. The remainder of the GABA-stained cells contain substantial levels of both GABA and glutamate, suggesting transmitter colocalization. In the toadfish utricle, glutamatergic hair cells are present throughout the macula. GABA-immunoreactive hair cells follow the arc of the striola, and most GABA-labeled receptor cells coexpress glutamate. The localization of GABA was explored in other species as well. In the pigeon, GABAergic hair cells are present throughout the crista ampullaris. Our findings demonstrate that multiple, neurochemically distinct types of hair cells are present in vestibular sensory epithelia. These observations, together with the excitatory activity generally associated with 8th nerve afferent fibers, strongly suggest that GABA serves an important, specific, and complex role in determining primary afferent response dynamics.

Cerebellar signatures of vestibulo-ocular reflex motor learning.

The vestibulo-ocular reflex (VOR) comprises an outstanding system to perform studies that probe possible cerebellar roles in motor learning. Novel VOR gains can be induced (learned) by the wearing of minifying or magnifying lenses, and learning requires the presence of the cerebellum. Previously, it was shown that Purkinje cells change their head velocity sensitivities with learning and that this change was thought to be inappropriate to be causal for the changed behavior. We now demonstrate that Purkinje cells also change their eye position, eye velocity, and head velocity sensitivities after learning. These combined changes at the Purkinje cell level contribute to a net modulation that is appropriate to support the new VOR gains. Importantly, the changes in the eye position parameter, reported for the first time, suggest the involvement of the neuronal integrator pathways in VOR learning. We provide evidence that all of these changes are necessary for VOR behavior and can explain learning deficits after cerebellar removal.

A dynamical model for the vertical vestibuloocular reflex and optokinetic response in primate.

The vestibuloocular reflex (VOR) in concert with the optokinetic response (OKR) stabilizes vision during head motion. The VOR system characteristics are both compensatory and adaptively self-calibrated. A model was constructed to aid in the understanding of the roles of the cerebellum and other neuronal sites in the performance and adaptation of the vertical VOR. The model structure was based upon the known neuroanatomy, and model parameters were estimated using experimental data. The model can reproduce and predict eye movements and cerebellar Purkinje cell firing patterns during VOR, OKR, and various visual-vestibular mismatch paradigms.