Responses of pigeon horizontal semicircular canal afferent fibers. II. High-frequency mechanical stimulation

1. The horizontal semicircular canals of anesthetized (barbiturate/ketamine) pigeons were mechanically stimulated by the use of a piezoelectric micropusher that provided controlled indentation of the surgically exposed membranous horizontal semicircular duct. 2. Extracellular action potentials from single horizontal semicircular canal afferent (HCA) fibers were recorded during sinusoidal mechanical stimulation. This method of stimulation was shown in the companion paper to produce equivalent responses to those produced by rotation for frequencies ranging from 0.01 to 10 Hz. 3. Sinusoidal mechanical stimulation produced clearly entrained action potentials in some HCA fibers up to a frequency of 400 Hz (highest stimulus frequency tested), with stimulus probe displacements of +/- 1.0 and +/- 2.5 microns. Thirty-four HCA fibers were thoroughly studied. 4. For most HCA fibers, the number of action potentials per stimulus cycle decreased as stimulus frequency increased, until only one action potential per stimulus cycle was elicited. The point at which only one spike per stimulus cycle was observed was dependent on both the fiber's resting mean discharge rate (MDR) and the fiber's coefficient of variation (CV) obtained during the MDR. 5. Dynamic response properties of individual HCA fibers were found to be correlated with the fiber's CV and the resting level MDR. Neurons with lower CV values had less adaptation, higher short time constants, and lower high corner frequencies than did neurons with high CV values. For a given CV class of HCA fibers, neurons with higher MDRs had more enhanced gains and more advanced phase shifts at high stimulus frequencies than did neurons with lower MDRs. 6. Transfer function parameters affecting the dynamics of the high-frequency response were derived from the mean gain and phase shift values of regular-, intermediate-, and irregular-firing HCA fibers. Best-fit short time constant (tau S) values of 4.6, 1.9, and 2.0 ms; hair cell membrane time constant (tau M) values of 10.3, 13, and 7 ms; excitatory postsynaptic membrane time constant (tau E) values of 0.8, 0.4, and 0.5 ms; and synaptic delay time constant (tau D) values of 0.5, 0.5, and 1.4 ms were determined for regular, intermediate, and irregular classes of HCA fibers, respectively. 7. The values of 4.6, 1.9, and 2.0 ms derived for the regular, intermediate, and irregular afferents would suggest upper-corner frequencies of 35, 84, and 80 Hz for these classes of HCA fibers, respectively.     

Influence of surgical plugging on horizontal semicircular canal mechanics and afferent response dynamics.

Mechanical occlusion of one or more of the semicircular canals is a surgical procedure performed clinically to treat certain vestibular disorders and used experimentally to assess individual contributions of separate canals and/or otoliths to vestibular neural pathways. The present experiments were designed to determine if semicircular canal afferent nerve modulation to angular head acceleration is blocked by occlusion of the endolymphatic duct, and if not, what mechanism(s) might account for a persistent afferent response. The perilymphatic space was opened to gain acute access to the horizontal canal (HC) in the oyster toadfish, Opsanus tau. Firing rate responses of HC afferents to sinusoidal whole-body rotation were recorded in the unoccluded control condition, during the process of duct occlusion, and in the plugged condition. The results show that complete occlusion of the duct did not block horizontal canal sensitivity; individual afferents often exhibited a robust firing rate modulation in response to whole-body rotation in the plugged condition. At high stimulus frequencies (about >8 Hz) the average sensitivity (afferent gain; spikes/s per degrees /s of head velocity) in the plugged condition was nearly equal to that observed for unoccluded controls in the same animals. At low stimulus frequencies (about <0.1 Hz), the average sensitivity in the plugged condition was attenuated by more than two orders of magnitude relative to unoccluded controls. The peak afferent firing rate for sinusoidal stimuli was phase advanced approximately 90 degrees in plugged canals relative to their control counterparts for stimulus frequencies approximately 0.1-2 Hz. Data indicate that afferents normally sensitive to angular velocity in the control condition became sensitive to angular acceleration in the plugged condition, whereas afferents sensitive to angular acceleration in the control condition became sensitive to the derivative of acceleration or angular jerk in the plugged condition. At higher frequencies (>8 Hz), the phase of afferents in the plugged condition became nearly equal, on average, to that observed in controls. A three-dimensional biomechanical model of the HC was developed to interpret the residual response in the plugged condition. Labyrinthine fluids were modeled as incompressible and Newtonian; the membranous duct, osseous canal and temporal bone were modeled as visco-elastic materials. The predicted attenuation and phase shift in cupular responses were in close agreement with the observed changes in afferent response dynamics after canal plugging. The model attributes the response of plugged canals to labyrinthine fluid pressure gradients that lead to membranous duct deformation, a spatial redistribution of labyrinthine fluids and cupular displacement. Validity of the model was established through its ability to predict: the relationship between plugged canal responses and unoccluded controls (present study), the relationship between afferent responses recorded during mechanical indentation of the membranous duct and physiological head rotation, the magnitude and phase of endolymphatic pressure generated during HC duct indentation, and previous model results for cupular gain and phase in the rigid-duct case. The same model was adjusted to conform to the morphology of the squirrel monkey and of the human to investigate the possible influence of canal plugging in primates. Membranous duct stiffness and perilymphatic cavity stiffness were identified as the most salient model parameters. Simulations indicate that canal plugging may be the most effective in relatively small species having small labyrinths, stiff round windows, and stiff bony perilymphatic enclosures.     

Discharge properties of afferent fibres of the goldfish semicircular canal with high frequency stimulation

The horizontal semicircular canals of goldfish were sinusoidally stimulated between 0.07 and 63 Hz (about 3 decades). Single afferent fibre recordings showed sinusoidal modulation of discharges. Above 4 Hz the discharges became phase-locked to the stimulus. With increasing frequency the number of spikes per period decreased so that finally only one spike per period remained. At 63 Hz a stimulus of as little as 0.005^o was sufficient to drive the units to far above their spontaneous activity. As cupular deflection is less than the angle of body movement, the cupular deflection threshold for modulation of afferent discharges must be much less than 0.005^o. Transfer functions of afferent activity were determined. The simple pendulum model does not fit the data. Additional introduction of a third time constant and the low pass properties of the receptor cell membrane, the synaptic delay and the leaky integrator of the post synaptic afferent terminal improve the fit.     

Behavior of horizontal semicircular canal afferents in alert monkey during vestibular and optokinetic stimulation

Summary The behavior of single vestibular nerve fibers from the lateral semicircular canal was recorded during sinusoidal oscillations of the head, during optokinetic stimulation with the head stationary, and during spontaneous oculomotor behavior in the alert monkey. The response of similar fibers to adequate vestibular stimulation was also studied in some of the animals under deeply anesthetized conditions. In the alert animals all units were spontaneously active and their discharge was modulated only by adequate vestibular stimulation. Ipsilateral horizontal rotations of the head were excitatory for all units. No modification of this basic vestibular response by visual stimulation including full-field striped drum rotation was observed. Furthermore no correlation of unit activity with oculomotor function including voluntary saccadic and pursuit eye movements was found in any of the units. The regularity of spontaneous discharge was the most consistent characteristic that differentiated the unit response into types. Most units were very regular in discharge, but a few were very irregular. The averaging of unit discharge over several cycles of oscillatory head rotation showed that the irregular type units were also consistently modulated by adequate vestibular stimulation. Both regular and irregular type units were found in the anesthetized animals. Unimodal distributions of the quantitative values for unit resting discharge rate, sensitivity, and phase relationship were found. The distributions for these three parameters were similar in the units recorded in the anesthetized animals. Thus at least these characteristics of semicircular canal response seem not to be affected by the vestibular efferent system which should be altered or eliminated in the case of the anesthetized animals.Research supported by NIH Grant EY0995-04.     

Morphological correlates of response dynamics and efferent stimulation in horizontal semicircular canal afferents of the toadfish, Opsanus tau.

1. We used the intraaxonal labeling technique to study correlations between the terminal dendritic morphology of horizontal semicircular canal primary afferents and their response dynamics to sinusoidal head rotation and combined electrical stimulation of central efferent vestibular neurons. Thirty-eight canal afferents were identified by their sensitivity and phase of response to rotation between 0.1 and 1.0 Hz (+/- 10 degrees/s) and were subsequently labeled with horseradish peroxidase or biocytin. The afferent's dendritic field and synaptic specializations in the neuroepithelium of the crista were examined under light microscopy. 2. Rate and regularity of background discharge of the afferent were not correlated with its axon diameter or relative location of its dendritic field in the crista. 3. Response sensitivity of the afferent to rotation was correlated both with the relative location of its dendritic field in the crista and with the number of terminal endings it possesses. Afferents having low sensitivities, slow dynamics, and few terminal endings supply the peripheral portions of the crista; afferents with higher sensitivities, faster dynamics, and greater number of terminal endings supply the more central portions. It is suggested that the differences in sensitivity among the afferents reflect principally the variations in both the cupular dynamics along the crista and the number of possible hair cell contact sites in the neuroepithelium. 4. Response phase of the afferent was correlated only with the extent of its dendritic processes along the transverse axis of the crista. Afferents having transversely oriented dendritic fields had less phase lags relative to acceleration than did those having a more longitudinally oriented dendritic field. 5. Efferent stimulation produced a change in both the afferent's discharge rate and its response sensitivity to rotation. Afferents discharge rate and its response sensitivity to rotation. Afferents having a centrally located dendritic field and acceleration afferents, defined by their response to rotation, were the most affected by efferent stimulation. These results suggest that efferent innervation is either directed toward, or most efficacious in, the central regions of the crista and that it may select specific hair cell-afferent complexes.     

Response characteristics of semicircular canal and otolith systems in cat. I. Dynamic responses of primary vestibular fibers

1. The activity of cat semicircular canal and otolith afferents was studied during yaw and roll rotations, respectively, to examine their dynamic behavior. 2. A sinusoidal analysis of the canal afferent activities showed that their dynamic characteristics are similar to those of second order vestibular neurons, except for a two to three-fold lower absolute gain. This agrees with earlier studies using angular acceleration steps. 3. Both divisions of the eighth nerve were sampled so as to examine afferents from both the utriculus and sacculus. Within the range of inputs used (+/- 25 degrees lateral tilt), the presumed sacular afferents (inferior division) showed either a gamma- or beta-response. However, the gain of their response was generally much less than for the afferents of the superior division (mostly utricular). This behavior is to be expected on the basis of receptor orientations and the components of gravity acting upon the macular receptors. 4. In response to ramp changes in angular position, some otolith units showed a phasic-tonic response pattern, i.e., an overshoot followed by an adaptation to a new steady state level of activity. The majority of units showed predominantly tonic responses proportional to displacement. 5. During sinusoidal rotations the predominantly tonic units showed small phase leads of 0 to 15 degrees at 0.025 Hz which remained constant or decreased to 0 to -15 degrees at 1.0 Hz. The gains were flat or increased by up to 2 fold. The phasic-tonic units showed greater phase leads, 10 to 50 degrees, and gains which increased from 2 to 8 fold. 6. This behavior of otolith afferents suggests that they can provide information about both the magnitude and the rate of change of linear acceleration stimuli.     

Spontaneous and driven responses of semicircular canal primary afferents in the unanesthetized pigeon

Spontaneous activity from 120 vestibular semicircular canal (SC) primary afferents and driven responses from 20 SC primary afferents to 2.3-decade bandwidth (0.029-6.152 Hz) sum of sinusoidal angular rotations were characterized in 10 unanesthetized pigeons. The results were compared with those previously gathered from barbiturate anesthetized pigeons. The average spontaneous mean firing rate (MFR) of SC primary afferents in unanesthetized pigeons was found to be 168 impulses per second (I/s), n = 120, and is approximately 80% higher than that for SC primary afferents in anesthetized pigeons [93 I/s, n = 149 (Ref. 9) and 92 I/s, n = 124 (Ref. 22)]. The spontaneous discharge of SC primary afferents from unanesthetized pigeons was classified according to the coefficient of variation (CV) of 512-1,024 interspike intervals (ISIs) into regular (CV less than 0.1), intermediate (0.1 less than or equal to CV less 0.4), and irregular (0.4 less than or equal to CV) categories. The percentages of SC primary afferents falling within each CV category were similar for unanesthetized and anesthetized pigeons (22). The relation between the spontaneous mean ISI and its standard deviation (SD) for SC primary afferents in the unanesthetized pigeon is best described by a power function model, which provides a significantly better fit than does a linear model. The parameters of this power function model are similar for SC primary afferents in unanesthetized and anesthetized pigeons (10). The spontaneous ISIs of individual SC primary afferents in the unanesthetized pigeon were found to be distributed either normally, log-normally, or according to the first passage time of the Wiener-Levy (WL) process. The gain of anterior SC primary afferents at 0.25 Hz is similar for anesthetized (2.93 I X s-1 X deg-1 X s-1, n = 14) (11) and for unanesthetized (3.01 I X s-1 X deg-1 X s-1, n = 14) pigeons. Bode plots constructed from the responses to sum sinusoidal angular accelerations reveal that SC primary afferents in unanesthetized pigeons display a phase lead and gain enhancement at frequencies between 0.6 and 6 Hz.(ABSTRACT TRUNCATED AT 400 WORDS)     

Studies of solitary semicircular canal hair cells in the adult pigeon. I. Frequency- and time-domain analysis of active and passive membrane properties

1. Hair cells were enzymatically dissociated from the neuroepithelium (cristae ampullares) of the semicircular canals of white king pigeons (Columba livia). Those hair cells determined to be type II by an anatomic criterion, the ratio of the minimum width of the neck to the width of the cuticular plate, were studied with the use of the whole cell patch-clamp technique. 2. The mean +/- SD zero-current membrane potential, Vz, was found to be -54 +/- 12 mV for anterior crista hair cells (n = 71), -62 +/- 14 mV for posterior crista hair cells (n = 14), and -55 +/- 12 mV for lateral (horizontal) crista hair cells (n = 18). The mean +/- SD value of Vz for hair cells from all cristae (n = 103) was -56 +/- 13 mV. 3. Active and passive membrane properties were calculated in the time domain, in voltage- or current-clamp mode, from responses to voltage or current pulses and, in the frequency domain, by fitting a membrane model to admittance magnitude and phase data resulting from current responses to sum-of-sines voltages at different d.c. levels of voltage-clamp membrane potential. 4. The average value +/- SE of input resistance (Rin), over the range from -100 to -60 mV, was found to 1.5 +/- 0.3 G omega from a mean-voltage-as-a-function-of-current plot, V-I, (n = 7) and a mean of 1.4 +/- 0.3 G omega from individual (n = 15) current-as-a-function-of-voltage plots, I-V. A lower mean value 0.8 +/- 0.4 G omega was obtained for the input resistance from frequency-domain calculations for a different set of cells (n = 21). Also, in two different sets of cells, average input capacitance (Cin) was determined to be 12 +/- 3 pF (n = 7) from time-domain estimates and 14 +/- 3 pF (n = 21) from frequency-domain estimates. The (Rin)(Cin) product was 11 ms based on frequency-domain estimates and 17 ms from time-domain estimates. 5. I-V curves for hair cells voltage clamped at -60 mV showed some anomalous rectification for hyperpolarizations between -60 and -120 mV but no detectable N-shape for depolarizations between -50 and 90 mV. The I-V relation showed increasing slope with depolarization through the resting potential (Vz) and increased linearly between -40 and 80 mV; the best-fit straight-line maximum slope conductance for six cells over this range was 17.4 +/- 0.3 nS.(ABSTRACT TRUNCATED AT 400 WORDS)     

Tachypnea after stimulation of afferent cardiac sympathetic nerve fibers.

The role of afferent cardiac sympathetic nerve fibers in the regulation of respiration has been examined. Application of potassium chloride or lactic acid solutions to the left ventricular surface of anesthetized vagotomized dogs resulted in a decrease in the manimum firing rate and shortening in period duration of firing of phrenic nerves. Also, application of the agents caused a decrease in amplitude and an increase in rate of respiratory thoracic movements. The same changes in phrenic nerve activity and respiratory movements were produced by coronary artery occlusion and centrifugal electrical stimulation of the left inferior cardiac nerves. The results indicate tachypnea that can be produced by excitation of afferent cardiac sympathetic nerve fibers.     

Studies of solitary semicircular canal hair cells in the adult pigeon. II. Voltage-dependent ionic conductances

1. The ionic conductances present in putative type II hair cells enzymatically dissociated from the anterior, posterior, and lateral semicircular canal cristae of the white king pigeon (Columba livia) vestibule were studied under whole cell voltage clamp. 2. Two classes of voltage-dependent potassium conductances were distinguishable on the basis of the time course of activation and inactivation and pharmacologic sensitivity. The rapid potassium conductance, IA, as inhibited by 6 mM 4-aminopyridine (4-AP), whereas the slow potassium conductance, IK, was inhibited by 50 mM tetraethylammonium (TEA). These conductances were not affected by extracellular calcium removal. IA was quite similar to the rapidly-inactivating A-current of molluscan soma, whereas IK was more like the delayed rectifier of molluscan soma. 3. The steady-state inactivation of IA occurred over a potential range from -100 to -40 mV. The threshold for activation of IA occurred between -60 and -50 mV. The slope conductance of the I-V curve over a range of -50 to -20 mV was 13.7 nS when the conditioning pulse was -100 mV, and we estimate it to be approximately 1-2 nS from the resting membrane potential of -56 mV. 4. The steady-state inactivation of IK was approximately 60% at -40 mV and was completely removed at -80 mV. The threshold for activation of IK was between -50 and -40 mV. The slope conductance of the I-V curve over a range of -50 to -20 mV was 10.5 nS when the conditioning pulse was -80 mV, and we estimate it to be approximately 6-7 nS from the resting potential of -56 mV. 5. At -56 mV (the average resting membrane potential of putative type II semicircular canal hair cells), approximately 10-14% of IA channels and approximately 57-70% of IK channels were not inactivated: thus IA and IK can contribute to the outward current during small depolarizations from rest. 6. A small calcium-dependent outward current, IK(Ca), could be elicited during step depolarizations from a holding potential of -40 mV. This calcium-dependent current was active over the range of -20 to +40 mV. 7. Inward currents could not be detected when the cells were exposed to normal physiological solutions. However, when the outward currents were blocked with internal cesium and the external solution contained 20 mM barium, sustained inward currents with rapid activation kinetics could be detected. The threshold for activation of the inward current occurred at -40 mV, and the I-V relationship peaked at -10 mV.(ABSTRACT TRUNCATED AT 400 WORDS)     

Opioid peptides as possible neuromodulators of the afferent synaptic transmission in the frog semicircular canal.

Vestibular receptors of the frog, Rana temporaria, were examined for the effect of bath-applied opioid peptide leu-enkephalin, its synthetic analogue dalargin and the specific opiate antagonist naloxone. Multiunit afferent activity of the whole vestibular nerve was recorded in an in vitro preparation. Leu-enkephalin (0.005-100 nM) and dalargin (0.1-100 nM) depress the resting discharge frequency. Naloxone (10 nM-1 microM) antagonizes responses induced by leu-enkephalin and dalargin that suggests a specific action of opioid peptides. Leu-enkephalin and delargin inhibit the excitatory action of L-glutamate. The effects of opioid peptides on L-glutamate-induced responses are unaffected by Co2+ block of transmitter release from hair cells that could speak in favour of the postsynaptic nature of these responses. At the same time, the other possible site of action of opioid peptides, such as efferent system, can not be excluded. The results indicate that opiate receptors are present in hair cells and that the neurotransmitter L-glutamate is involved in opiate action at the peripheral vestibular system of the frog. We suggest that opioid peptides may act as a neuromodulator in this system.     

Frequency-response analysis of vestibular-induced neck reflex in cat. I. Characteristics of neural transmission from horizontal semicircular canal to neck motoneurons.

1. Vestibular-induced neck muscle reflexes (vestibulocollic reflexes) were studied with frequency-response methods in unanesthetized, decerebrate cats. The horizontal semicircular canals were stimulated by oscillation of the turntable and EMG activity was recorded from neck extensor muscles. 2. The maximum firing rate of each motor unit was less than 40 spikes/s. The motor units were classified by their maximum firing rates into two groups: HF (high frequency) units which could fire above 20 spikes/s and LF (low frequency) units which could not exceed 20 spikes/s of firing. The HF units had larger gains than the LF units on the average. 3. Compound EMGs, which presumably represent activity of the whole muscle, were examined at different frequencies of stimulation. The gain of compound EMG responses depended on the spontaneous activity. When the spontaneous activity was low or too high, the gain was small. There was an intermediate spontaneous activity level at which the gain became maximal. 4. The phase lag and the gain of the frequency response were represented in a Bode diagram with respect to angular acceleration. The transfer function of the system from the vestibular nuclei to neck EMG activity was estimated by assuming a first-order lag-lead system. A phase lag of 70-80 degrees (0.05 Hz) was found between the averaged activity of vestibular nucleus neurons and cervical motor activity. A positive correlation was observed between the phase lag and the gain of each motor unit. These results cannot be attributed solely to the action of the vestibulospinal tract, but suggest the existence of a neural intergrator in the vestibulocollic reflex arc.     

Directionally-specific effects of afferent signals from the extraocular muscles upon responses in the pigeon brainstem to horizontal vestibular stimulation

The responses of single units in the brainstem of the decerebrate, paralysed, pigeon were studied. Natural vestibular stimulation was provided by horizontal, sinusoidal, oscillation of the bird and extraocular muscle afferents of the ipsilateral eye were activated by passive eye-movement. Unit responses to vestibular and/or orbital stimuli were examined in sets of peristimulus time histograms interleaved in time. Of 352 units in the brainstem, in the region of the vestibular nuclei, which were exposed to the effects of both vestibular stimuli and passive eye-movement, 40 (11%) responded only to the latter; the other 312 units (89%) responded to vestibular stimulation at 0.4 Hz (amplitude +/- 8 degrees). Of these 312 units, 129 (41%) were affected only by vestibular stimuli; in the other 183 units (59%) passive eye-movement produced clear modification of the vestibular responses by adding excitation or inhibition, or both. There were phasic modifications in most units; in 77 there were longer-lasting changes in the vestibular responses, often following a phasic response. In 124 units whose responses were subjected to statistical analysis, the vestibular responses of 42 (34%) were modified only by horizontal eye-movement and eight (6%) were affected only by vertical movement. A further 18% showed larger effects from horizontal than from vertical eye-movement; in 2% vertical eye-movement was preferred. Further examination of the specificity of the effects of eye-movement in planes between the vertical and horizontal was possible in 29 units which showed various degrees of "tuning" of the effect. In some units there was additional specificity for eye-movement in (a) particular directions (towards the beak rather than towards the tail, for example); (b) in particular arcs of the orbit (centre-to-temporal rather than nasal-to-centre, for example). Note that all these effects were upon the responses of the units to horizontal vestibular stimulation. Thus, the modifications of the vestibular responses depended upon specific characteristics of the passive eye-movement. The exact recording sites of 29 units were determined histologically; some were in the medial vestibular nucleus but many were in the adjacent reticular formation. The principal interest of the results is that they provide more detailed information than was available previously on the specificity of the effects of afferent signals from the extraocular muscles upon the vestibular responses of units in regions of the brainstem known to be involved in oculomotor control. The decerebrate pigeon proves to be a particularly good preparation in which to study these effects.(ABSTRACT TRUNCATED AT 400 WORDS)