Michael J. Lyon

Localization of adductor and abductor motor nerve fibers to the larynx.

Knowledge of the location of motor nerve fibers to the adductor and abductor muscles of the larynx may be useful in the diagnosis and treatment of innervation disorders in this organ. Anterograde degeneration and retrograde tracer anatomical techniques have demonstrated the central and peripheral positions of these two groups of motor nerve fibers in the cat. Traditional nerve fiber degeneration methods applied following intracranial transection of the vagus nerve rootlets indicated that: 1) Most of the fibers in the recurrent laryngeal nerve (RLN) are motor; 2) Almost all of these motor fibers leave the brain stem in the most rostral rootlet(s) of the vagus nerve; and 3) Motor fibers to the larynx form a discrete bundle within the trunk of the vagus nerve before forming the RLN. A tracer (horseradish peroxidase) of retrograde axoplasmic flow in motor neurons has been employed to demonstrate: 1) Dorsoventral division of the adductor and abductor neurons in the nucleus ambiguus; and 2) Diffuse arrangement of both adductor and abductor nerve fibers in the vagus nerve but collection of these fibers into abductor and adductor halves of the RLN prior to entering the larynx. These findings dispel theories of differential cord paralysis (Semons law) based on a vulnerable position of abductor fibers at the periphery of the RLN. Furthermore, the diffuse arrangement of these fiber groups explains the usually mixed functional results obtained following reimplantation of the RLN into a laryngeal muscle.

Fiber Components of the Recurrent Laryngeal Nerve in the Cat

Experimental neuroanatomical methods were employed in 21 adult cats to determine 1) the number and size of myelinated motor and sensory fibers in the recurrent laryngeal nerve (RLN), and 2) the fiber components originating in the nucleus ambiguus (NA) and retrofacial nucleus (RFN) of the brain stem. Intracranial transection of the X and XI cranial nerves and selective destruction of the NA or RFN were the experimental lesions inflicted in order to obtain the following results. About 55% (312) of the right RLN (565 fibers) is composed of myelinated motor nerve fibers which measure 4 μ − 9 μ in diameter. Nine percent come from the RFN and are smaller (4–6 μ) than the 46% which emanate from the NA and measure 6–9 μ in diameter. The remaining 45% of the RLN is made up of sensory neurons which can be divided into three groups. 1)The largest numerical group (32%) is very small in caliber (1–3 μ) and supplies extralaryngeal regions (trachea, esophagus). 2) The intermediate size fiber group (4–9 μ) comprises 11% of the RLN and probably supplies the subglottic mucosa. 3) The smallest group (2%) of sensory fibers is the largest in diameter (10–15 μ) and may represent either the innervation of muscle spindles or afferents from the superior laryngeal nerve coursing down into the chest.

Ultrastructural Changes in Vestibulo-Ocular Neurons following Vestibular Neurectomy in the Cat

Vestibulo-ocular (VO) neurons in the superior vestibular nucleus were labeled retrogradely with horseradish peroxidase and studied quantitatively using electron microscopy to determine the morphologic correlates of vestibular compensation. Eleven VO neurons from three normal cats were compared to 26 VO neurons in four animals killed 8 weeks after vestibular neurectomy and 13 VO cells from two animals killed 1 year after vestibular neurectomy. The results demonstrated a marked reduction (74%) in the number of synaptic profiles (SPs) on the VO cell soma in both experimental groups. Synaptic vesicles in the remaining SPs on VO neurons were fewer, smaller, and rounder than vesicles in control animals. The residual SPs also were associated with more asymmetric synapses. The experimental VO neuron showed a significant decrease in soma and organelles associated with protein synthesis.

Blood flow and assessment of capillaries in the aging rat posterior canal crista

Vascular change has been proposed as an etiological factor in inner ear aging and in several inner ear disorders. Moreover, some successful medical management of the episodic vertigo and tinnitus associated with Ménières disease has been directed toward pharmacologically increasing blood flow, changing vascular permeability or ion homeostasis. While there are many studies of cochlear capillary morphology and blood flow, there are very few examining these variables in the vestibular system and none with respect to aging. The purpose of this study was to examine the rat posterior canal ampullary crista for age-related changes in blood flow and capillary morphology. By combining stereological techniques with microsphere injection, we have determined that in the rat posterior canal crista there is a statistically significant age-related decrease in blood flow (75%), mean capillary diameter (31%), and volume fraction of capillary lumen (31%). There is also an overall 18% decrease in the volume of the ampullary crista, a 72% decrease in blood flow/unit volume and a 36% increase in capillary length/unit volume. There were no significant changes in the capillary surface area/unit volume, the absolute capillary length, or the absolute capillary surface area. These data suggest impaired blood flow and degenerative loss of the ampullary crista may be relate to impaired end organ function.

Aging rat vestibular ganglion: I. Quantitative light microscopic evaluation

This study was undertaken to quantify age-related changes in the rat vestibular ganglion. Cell number, diameter, and proximal-distal distribution based on size were evaluated. Serial 5-microns plastic sections of the vestibular ganglion from 15 female Wistar rats were examined. Rats were divided into three age groups: young (Y, 3 to 5 months, n = 5), old (0, 24 to 26 months, n = 3), and very old (VO, 28 to 31 months, n = 7). Quantitative analysis indicated no significant differences (P less than .05) in the estimated number of ganglion cells (mean: Y = 1,690, 0 = 2,257, VO = 1,678), ganglion cell profile diameters (mean: Y = 22.5 microns, n = 2,886; O = 23.7 microns, n = 2,313; VO = 22.8 microns, n = 4,061), or proximal-distal localization (proximal: 22.3 microns, 24.4 microns, 22.7 microns; middle: 22.6 microns, 23.1 microns, 22.4 microns; distal: 23.3 microns, 23.4 microns, 23.7 microns; Y, O, and VO, respectively). When pooled, the old animals tended to have slightly larger cell profiles than the other groups. We noted a dramatic age-related increase of aging pigment within the ganglion cell profiles, making the old and very old animals easily distinguishable from the young. In most of the cell profiles, the aging pigment was more or less uniformly distributed throughout the cytoplasm. However, in some, aging pigment was accumulated at one pole of the cell profile. While no typical degenerating cellular profiles were found in any of the sections, several of the ganglion cell profiles from the old animals revealed dense cytoplasm, possibly indicating an early stage of degeneration.

Aging Rat Vestibular Ganglion: II. Quantitative Electron Microscopic Evaluation

This laboratory has shown that age-related vestibular ganglion cell loss does not occur in the Wistar rat as it does in humans. However, in that study, intracellular changes were evident. The purpose of the present study was to quantitate some of these changes. The volume densities of mitochondria, rough endoplasmic reticulum (RER), Golgi apparatus, and aging pigment, as well as the diameter of the vestibular ganglion cells, of young (3 to 5 months) and old (24 to 31 months) female Wistar rats were determined by electron microscopy and stereological techniques. The data show a significant decrease in the volume densities of mitochondria (11.4%), Golgi apparatus (8.1%), and RER (8.9%), a significant increase in aging pigment (327%), and no change in mean profile diameter. These results suggest a decreased capacity for oxidative metabolism and protein synthesis that may reflect a decrease in the number of hair cells innervated by each ganglion cell and/or in the number of central connections. In either case, these findings suggest impaired metabolic and functional capabilities.

Morphologic correlates of vestibular compensation in the cat.

Vestibulo-ocular (VO) neurons in the superior vestibular nucleus (SVN) were labeled with horseradish peroxidase (HRP) and studied quantitatively in the electron microscope to determine the morphologic correlates of vestibular compensation. Eighteen cells from four normal cats were compared to 26 cells from four 8-week vestibular neurectomy animals and 29 cells from four 1-year neurectomy animals. There was a greater than 70% decrease in synaptic profiles (SP) on the VO soma in both experimental groups. Increased excitatory activity in the residual SP are indicated by (1) smaller synaptic vesicles (SV) with increasing survival after neurectomy, (2) the association of the SP with a higher ratio of asymmetric to symmetric synapses, and (3) a higher volume fraction of mitochondria in the SP. The experimental VO soma showed a 17% reduction in size (cytoplasm), and a decrease in rough endoplasmic reticulum and polyribosomes. The target cell also showed an increased number of somal spines associated with the remaining SP in the 1-year neurectomy group.

Central projection of the sensory component of the rat recurrent laryngeal nerve

The central projection from the sensory components in the rat recurrent laryngeal nerve was studied using WGA-HRP. Sensory terminals were found bilaterally in the nuclei of the tractus solitarius, although they were very sparse contralaterally. In the ipsilateral nucleus, most of these terminals were located in the interstitial subnucleus extending, from the most rostral area, near the obex, caudally for a distance of 1.5 mm.

Ultrastructural Changes in Contralateral Ves tibulo-ocular Neurons Following Vestibular Neurectomy in the Cat

Twenty contralateral superior vestibular vestibulo-ocular neurons (SVON) from 4 cats were studied morphologically 8 weeks after a right vestibular neurectomy. Nine SVON demonstrated a 35% loss of somal synaptic profiles (SP), normal nuclear and cytoplasmic size, but a slight decrease in organelles responsible for protein synthesis (rough endoplasmic reticulum and polyribosomes). Eleven SVON showed an 82% loss of SP, decrease in cell and nuclear size, and significant reduction in rough endoplasmic reticulum (RER) and polyribosomes. These transneuronal SVON changes are presumed to be the result of commissural pathway degeneration caused by the vestibular neurectomy.

Comparison of the vascular innervation of the rat cochlea and vestibular system.

In order to gain a better understanding of the neuronal and local control of inner ear blood flow, the vascular innervation to the rat cochlea and vestibular system was examined. Specimens were removed in toto beginning at the basilar artery extending to the anterior inferior cerebellar artery, labyrinthine artery, common cochlear artery, modiolar artery and anterior vestibular artery. When possible the vessels were dissected in continuity through the cribrose area. The vestibular endorgans were also removed. Specimens were examined using immunohistochemical techniques for the presence of vasoactive intestinal peptide, neuronal nitric oxide synthase, neuropeptide-Y, substance P and calcitonin gene related peptide. Results show that the vasculature to the cochlea and vestibular portion of the inner ear receive similar types of nonadrenergic innervation, that within the vestibular endorgans, only CGRP and SP were found in the neuroepithelium or in association with vessels, and that within the vestibular system, the majority of the vascular innervation appears to stop at or near the cribrose area. In the cochlea however, it extends to include the radiating arterioles. These findings suggest that cochlear blood flow is under finer control and that neuronally induced changes in blood flow may have a more global effect in the vestibular periphery.