Karl Beykirch

Comparison of static and dynamic posturography in young and older normal people

Objective: To measure sway velocity during static and dynamic posturography in “normal’ older people and to determine which tests best distinguish young from older subjects.

Quantification of the process of hair cell loss and recovery in the chinchilla crista ampullaris after gentamicin treatment.

The degree of ototoxic drug sensitivity and hair cell repair was determined in the chinchilla horizontal crista ampullaris after intraotic administration of gentamicin. Histological evaluation was made of 22 cristae ampullaris from one normal and six post‐treatment (PT) animal groups killed at 1, 4, 7, 14, 28, and 56 days. New hair cell production was quantified, using the dissector technique. Transmission electron microscopy was used to investigate the ultrastructural characteristics of the hair cells in the regenerated epithelium. At I day PT, type I and II hair cells presented cytoplasmic vacuolization, swollen nerve calyces and 20% of type I and 18% of type II hair cells were lost. At 4days PT, 95% of type I hair cells and 14% of type II hair cells had disappeared. In addition, most of the type II hair cells showed clumping of nuclear material. Nerve fibers were not found in the sensory epithelium, but were still observed below the basal lamina. Supporting cells appeared unaffected, maintaining their location in the crista. At 1 and 4 days PT, the damage to hair cells was more pronounced in the central region of the crista ampullaris. The degree of ototoxic damage at 7 days was similar to that of 14 days: no type I hair cells were present and most of the type II hair cells had disappeared; supporting cell nuclei began to occupy the apical part of the sensory epithelium and most of the nerve fibers had retracted. Quantitatively, 87 and 93% of type II hair cells were lost at 7 and 14 days PT, respectively. Initial signs of hair cell recovery began at 28 days PT; immature type II‐like hair cells appeared, supporting cell nuclei began to align at the base of the sensory epithelium and nerve fibers penetrating the basal lamina were observed. No type I hair cells were found, but 40% of the normal number of type II hair cells were present. Hair cells appeared to regenerate in the peripheral areas of the cristae ampullaris first. At 56 days PT, an increase in the number of mature type II hair cells was present, supporting cells were aligned at the base of the epithelium, and more nerve fibers appeared to penetrate the basal lamina to the sensory epithelium. Although type I hair cells were absent from the epithelium, 55% of the normal number of type II hair cells were present. At this time, more regenerated hair cells were located in the center of the cristae ampullaris as compared to the periphery. At the transmission electron microscopic level, type II hair cells at different stages of maturation were observed. Some exhibited mature stereocilia, a cuticular plate, and terminal endings with synaptic specialization opposing these hair cells. In conclusion, type I hair cells were more sensitive than type II hair cells to gentamicin intoxication (as they disappeared as early as 4days PT). After 56 days PT, the number of type II hair cells reached 55% of normal. No type I hair cells had regenerated at this time. These results demonstrate quantitatively the differential ototoxic sensitivity and regenerative capacity of hair cells.

Projections of the individual vestibular end-organs in the brain stem of the squirrel monkey

The central nervous system (CNS) projections of primary afferent neurons from individual vestibular receptors were studied using horseradish peroxidase (HRP) or biocytin labeling in 14 ears from 7 adult squirrel monkeys using the technique developed in the chinchilla (Lee et al., 1989, 1992). The specificity of labeling was verified by examining the location of the labeled fibers and cell bodies in the vestibular nerve and Scarpas ganglion. Labeled fibers and cells were restricted to nerves and areas belonging to groups of cells in either the superior or the inferior ganglion of the vestibular nerve. In the vestibular nerve root, labeled primary afferent fibers also exhibited a receptor-dependent segregation at the entrance to the medulla. Fibers from the HSC and the SSC were found rostrally and those from the PSC and the SAC were found in the caudal area. The UTR fibers were situated intermediate between these two groups of fibers. (A bundle of fibers, probably vestibular efferents, was identified immediately rostrally and ventromedially to the UTR fibers.) The primary afferent fibers bifurcated into secondary ascending and descending fibers at the lateral border of the vestibular nuclei, forming a longitudinal rostrocaudal vestibular tract. The secondary fibers from individual end-organs occupied specific locations in the tract: the UTR fibers were dorsal to the SSC and the HSC fibers, PSC fibers were found most medially, and the SAC fibers occupied the lateralmost area. The secondary UTR fibers overlapped considerably with those of the SSC and the HSC. The orderly receptor-dependent segregation of fibers was more prominent in the descending tracts than in the ascending tracts. In the vestibular nuclei complex the location of the tertiary branches of various end-organs exhibited considerable overlap within the major vestibular nuclei (SN, superior nucleus; LN, lateral nucleus; MN, medial nucleus; DN, descending nucleus). There were still differences, however, in the projection pattern. Fibers from the SAC ran primarily in the lateral area, fibers from the SSC and the UTR were found ventromedially to the SAC fibers, and the HSC projected slightly medially to the fibers from the SSC. The PSC fibers projected most medially. The UTR and SAC sent numerous fibers to the cerebellum. Fibers from the semicircular canals projected through the rostrodorsal region of the SN and presumably also projected to the cerebellum. The precise termination of fibers was evaluated by studying the location of labeled boutons, which were identified in all major vestibular nuclei. Labeled boutons from all the receptors were in the rostral and central areas of the SN, and in the MN mainly in the rostral two-thirds. In the LN, boutons from all the receptors were in the rostroventral part, most of which were from the UTR and SAC. No labeled boutons were in the caudodorsal part of this nucleus. Labeled boutons in the DN primarily surrounded the descending tract fibers and were particularly prominent medially. In specimens in which superior vestibular nerve receptor organs were scratched vestibular efferent fibers were also labeled. These fibers traveled in the most ventral part of the vestibular nerve root and projected in the ventral aspect of the LN to labeled soma in the ipsilateral and contralateral brain stem. Specificity the in projection patterns of efferent fibers from different end-organs could not be ascertained.

HAIR CELL RECOVERY IN THE CHINCHILLA CRISTA AMPULLARIS AFTER GENTAMICIN TREATMENT : A QUANTITATIVE APPROACH

The mechanisms for hair cell recovery were investigated after intraotic application of 50 μg gentamicin into the perilymphatic space of the superior semicircular canal of the chinchilla. Histologic evaluation of one normal group and four posttreatment groups (7, 14, 28, and 56 days) was made with light and transmission electron microscopic techniques. The numeric changes of hair cells and supporting cells was quantified with the dissector technique. At 7 and 14 days after treatment, no type I hair cells were present, and 85% and 88% of type II hair cells were lost. Supporting cells decreased to 76% at 7 days, but they recovered to 91% at 14 days. Recovery of the epithelia was evident 28 days after treatment; 83% were type II hair cells, and 3% were type I hair cells. The supporting cell number remained close to normal (86%). Between 14 and 28 days after treatment, there was an increase of 1758 of type II hair cells, representing approximately 125 new hair cells per day. At the same time interval the number of supporting cells remained near normal. These results suggest that new hair cells might be the result of supporting cell mitotic division and differentiation.

Horizontal Vestibulo-ocular Reflex after Acute Peripheral Lesions

We studied the dynamics of the horizontal vestibulo-ocular reflex using a broad range of sinusoidal rotational stimuli in 14 patients with acute unilateral peripheral vestibular lesions. From plots of slow phase velocity (SPV) versus stimulus velocity (phase shifts removed) we calculated the DC offset and gain to ampullopetal (AP) and ampullofugal (AF) stimulation of the remaining intact labyrinth. DC offset values were highly correlated with spontaneous nystagmus SPV values for all frequencies (0.0125-0.8 Hz) and amplitudes (30-120-deg/sec) of stimulation. Mean AP gain values were consistently higher than mean AF values with the greatest difference occurring at low frequencies of stimulation. Saturation nonlinearities were present with AF stimulation at high amplitudes. These findings can be explained by dynamic asymmetries at the level of the primary and secondary vestibular neurons.

The Electrically Evoked Vestibulo-Ocular Reflex: I. Normal Subjects

Recent animal studies indicate that electric currents applied through perilymphatic-space electrodes stimulate vestibular primary afferent neurons directly. These findings suggest that electrical stimulation may provide a testing method by which the vestibular nerve and central pathways could be evaluated separately from the vestibular end-organ. The goal of this study was to obtain normative data on human beings for an electrically evoked vestibulo-ocular reflex (EVOR). Sinusoidal electrical stimuli (0.0125 to 0.8 Hz, 4 mA peak intensity) were applied along the interaural axis through mastoid electrodes in 10 subjects. Horizontal eye movements were recorded by an infrared limbus-tracking device. The subjects also underwent rotational stimulation at the same frequencies so that their horizontal vestibulo-ocular reflex (VOR) could be evaluated. Nystagmus was observed in the EVOR at lower stimulus frequencies, whereas purely sinusoidal eye deviations occurred at higher frequencies. The phase of the EVOR slow-component eye velocity consistently lagged the stimulus. This contrasts with the phase measurements of the VOR in the same subjects, which exhibited a lead relative to head velocity. These findings suggest that currents applied to human beings may activate vestibular primary afferents independent of peripheral receptor mechanisms and thereby provide a “site-of-lesion” testing method by which the vestibular nerve and central pathways can be evaluated separately from the vestibular end-organ.

Arrangement of Vestibular Nerve Fibers in the Semicircular Canal Crista of the Chinchilla

The topographic arrangement of vestibular nerve fibers innervating semicircular canal cristae of the chinchilla was studied using computer-aided video-microscopy and three-dimensional reconstruction. At the level 20 microns proximal to the base of the crista, bundles consisting of 30-50 nerve fibers each were identified. Nerve fibers in bundles were classified into seven categories depending on the diameter. We confirmed that large nerve fibers were more frequently found in the central bundles and small nerve fibers were more frequently found in the peripheral bundles. The central bundle might function as a physiological unit coding various types of head movements, whereas the peripheral bundle might contribute more to the detection of slow and long-lasting movements giving rise to tonus and posture changes. The canalicular nerve may code rotational acceleration of the head via function- and locus-specific nerve fiber bundles.

Sensoritopic and Topologic Organization of the Vestibular Nerve

Anatomical studies in the vestibular system represent one of the earlier steps of Professor Rafael Lorente de No’s search for understanding of brain function. His investigations into the anatomy and physiology of the vestibular system brought the attention of the neuroscience community to this very young (early 20’s), curious, creative and tenacious worker, one of the last students of Nobel Laureate Santiago Ramon y Cajal. Lorente de No made many contributions which remain the foundation of vestibular science, such as the well-known descriptions of the anatomy and physiology of the three neuron vestibuloocular reflex arc and interneurons in vestibular reflex function (Lorente de No,’ 33b; Lorente de No,’ 38).

The galvanically-induced vestibulo-ocular reflex in the cat

Rotatory vestibular input is processed by receptor organs and relayed along the vestibular nerves to a central processor, whence the vestibulo‐ocular reflex (VOR) is generated. Electrical stimuli applied to the labyrinth can bypass receptors and stimulate the nerve directly, thereby generating horizontal eye movements (EVOR). It was our purpose to mathematically relate the EVOR to the VOR in an effort to generate a parameter by which the experimental effects of ototoxins on the inner ear can be evaluated.