Ultrastructural histopathology of human olfactory dysfunction.

This paper presents electron-microscopic observations on biopsies of the olfactory mucosae of several classes of patients with smell disorders: 1) patients with loss of smell function following head injury (post-traumatic anosmics or hyposmics); 2) patients with loss of smell function following severe head colds and/or sinus infections (post-viral olfactory dysfunction, or PVOD); and 3) patients that have lacked smell function since birth (congenital anosmics). Of these, the traumatic anosmics' olfactory epithelia were quite disorganized; the orderly arrangement of supporting cells, ciliated olfactory receptor neurons, microvillar cells, and basal cells was disrupted. Although many somata of ciliated olfactory receptors were present, few of their dendrites reached the epithelial surface. The few olfactory vesicles present usually lacked olfactory cilia. The post-viral anosmics, too, had a greatly reduced number of intact ciliated olfactory receptor neurons, and most of those present were aciliate. The post-viral hyposmics had a larger population of intact, ciliated olfactory receptor cells. In the seven cases of congenital anosmia studied, no biopsies of olfactory epithelium were obtained, indicating the olfactory epithelium is either absent--or greatly reduced in area--in these individuals.

Ultrastructural neurobiology of the olfactory mucosa of the brown trout, Salmo trutta.

This paper describes four investigations of the olfactory mucosa of the brown trout: 1) the ultrastructure of the olfactory mucosa as revealed by scanning (SEM), conventional transmission (TEM), and high voltage (HVEM) electron microscopy; 2) light and electron-microscopic investigations of retrograde transport of the tracer macromolecule horseradish peroxidase (HRP) when applied to the cut olfactory nerve; 3) SEM and TEM investigations of the effects of olfactory nerve transection on cell populations within the olfactory epithelium; and 4) ultrastructural investigations of reversible degeneration of olfactory receptors caused by elevated copper concentrations. The trout olfactory epithelium contains five cell types: ciliated epithelial cells, ciliated olfactory receptor cells, microvillar olfactory receptor cells, supporting cells, and basal cells. The ciliated and microvillar olfactory receptor cells and a small number of basal cells are backfilled by HRP when the tracer is applied to the cut olfactory nerve. When the olfactory nerve is cut, both ciliated and microvillar olfactory receptor cells degenerate within 2 days and are morphologically intact again within 8 days. When wild trout are taken from their native stream and placed in tanks with elevated copper concentrations, ciliated and microvillar cells degenerate. Replacement of these trout into their stream of origin is followed by morphologic restoration of both types of olfactory receptor cells. Ciliated and microvillar receptor cells are primary sensory bipolar neurons whose dendrites make contact with the environment; their axons travel directly to the brain. Consequently, substances can be transported directly from the environment into the brain via these "naked neurons." Since fish cannot escape from the water in which they swim, and since that water may occasionally contain brain-toxic substances, the ability to close off--and later reopen--this anatomic gateway to the brain would confer a tremendous selective advantage upon animals that evolved the "brain-sparing" capacity to do so. Consequently, the unique regenerative powers of vertebrate olfactory receptor neurons may have their evolutionary origin in fishes.

The vomeronasal (Jacobson's) organ in man: ultrastructure and frequency of occurrence.

These investigations address three major questions: (1) What is the frequency of occurrence of the vomeronasal (Jacobson's) organ (VNO) in man? (2) what is the ultrastructure of the human VNO? and (3) does the VNO contain sensory receptor cells? Macroscopic and microscopic intranasal clinical examinations of over 200 persons revealed paired bilateral vomeronasal pits on the anterior 1/3 of the nasal septum in all cases. Biopsies of the vomeronasal pits and surrounding tissues were examined by light and electron microscopy. These studies showed that the vomeronasal pit leads to a closed tube, 2-8 mm long, lined by a unique pseudostratified columnar epithelium unlike any other in the human body. The anterior end of the tube is lined by tall, columnar cells with a sparse population of short microvilli. The posterior end of the VNO is lined by an epithelium that contains three morphologically distinct cell types: (1) basal cells; (2) "dark cells--tall, slender cells with heterochromatic nuclei and electron-dense cytoplasm that often contain mucigen-like granules; and (3) "light" cells--large, clear cells, extending from the basement membrane to the organ's lumen. Each "light" cell has a round, euchromatic nucleus and a clear cytoplasm that often contains many Golgi stacks and membrane-limited vesicles filled with material of modest electron density. The cell apex is tipped by a few short microvilli. Whether these cells subserve any sensory function awaits further investigation.

The effect of chronic cocaine abuse on human olfaction.

Cocaine has been used for many decades as both a stimulant and as a topical anesthetic/vasoconstrictor. Illicit "snorting" or freebase smoking has increased markedly in recent years. Decreased olfaction has been an often reported subjective complaint of cocaine abusers, but quantification of smell loss using sensitive psychophysical tests has not yet been done, leading to the present study. Eleven cocaine abusers were recruited from a drug treatment clinic. Olfaction was assessed using a butanol threshold test, the UPSIT, and a 7-item discrimination test. One patient tested anosmic, one had a mild discrimination problem, and one had a large septal perforation but was normosmic. From the present study, it appears that most cocaine abusers, even heavy users or those with intranasal damage, do not develop permanent olfactory dysfunction. It is not clear what factors may have resulted in complaints of olfactory loss in previous studies.

Congenital anosmia.

Seven patients with congenital anosmia underwent detailed chemosensory evaluation, followed by the performance of biopsies of the olfactory region. Olfactory epithelium was not found in any of the biopsy specimens. It appears therefore that patients with congenital anosmia lack any olfactory epithelium. Several possible explanations for this finding are discussed. The most attractive hypothesis is that the olfactory placode forms either normally or abnormally during development but later degenerates and is replaced with respiratory epithelium. Only one patient in our series had congenital anosmia in association with a syndrome (Kallmann's syndrome), indicating that congenital anosmia is found more often as an isolated symptom.

Peroxidase backfills suggest the mammalian olfactory epithelium contains a second morphologically distinct class of bipolar sensory neuron: the microvillar cell.

The olfactory epithelium of man, rat and some other mammals consists of 4 cell types: ciliated olfactory receptors, microvillar cells, supporting (sustentacular) cells, and basal cells. Of these, the microvillar cell is least well understood: its function is unknown. In this study, a hypothesis is put forth: that the microvillar cells in the mammalian olfactory epithelium comprise a morphologically distinct class of sensory receptor. The hypothesis is tested by injecting the cytochemical tracer macromolecule horseradish peroxidase (HRP) into the olfactory bulb of the rat, and observing its pattern of uptake in the olfactory epithelium by light and electron microscopy. In these experiments, ciliated olfactory receptors and microvillar cells backfilled with HRP: supporting and basal cells did not. The data, which support the hypothesis, indicate the microvillar cells, along with the ciliated olfactory receptors, send axons to the olfactory bulb. Consequently, it is concluded that the microvillar cell is a sensory bipolar neuron, with the cell body in the olfactory epithelium, that sends a dendrite to the site of stimulus reception at the free surface of the olfactory epithelium, and an axon to the olfactory bulb in the brain. The similarity of microvillar cells in the olfactory epithelium to 'brush cells' found throughout the respiratory tract is discussed in detail.

Post-traumatic anosmia. Ultrastructural correlates.

Five patients suffering post-traumatic anosmia were studied at the University of Colorado Health Sciences Center, Denver. Each patient underwent psychophysical testing, clinical evaluation, and olfactory biopsy. The biopsy specimens were examined ultrastructurally and were found to vary from normal tissues. The overall appearance of the olfactory epithelium in the post-traumatic patient is disrupted and the receptor cells are distorted. Large numbers of axons are located near the basement membrane and can often be found in bundles throughout the epithelium, extending even to the mucosal surface. Olfactory cilia are rarely seen in epithelia obtained from post-traumatic patients. Bald olfactory vesicles, often containing basal bodies, are frequently observed. We postulate that in these cases, the olfactory epithelium regenerates following head trauma and the receptor cells attempt to send axons centrally. However, the cribriform plate has undergone fibrotic healing and the axons are unable to penetrate it and make contact with olfactory bulb neurons.

Steroid-dependent anosmia.

In steroid-dependent anosmia (nasal polyps, inhalant allergy, anosmia), high doses of steroids will temporarily restore the sense of smell, a diagnostic test. Appropriate surgery can then be carried out, followed by low-dose, long-term steroid therapy to maintain the sense of smell. Olfactory biopsy specimens taken during the course of evaluation and treatment show electron-optically normal olfactory receptors, meaning that the probable pathogenesis of the sensory deficit is an obstruction, mechanical and possibly biochemical. Two cases of steroid-dependent anosmia are presented to detail a fully reversible anosmia using state-of-the-art techniques.

Electron microscopy of olfactory epithelia in two patients with anosmia.

Ultrastructural alterations were present in biopsy specimens of olfactory epithelia taken from two patients with anosmia. In both cases, the olfactory epithelia presented a disorganized appearance when viewed by transmission electron microscopy. The number of ciliated olfactory receptors was reduced; few olfactory vesicles were present at the epithelial surface. Where present, the olfactory vesicles usually lacked cilia. Since both patients had a history of head trauma, we speculate that the fila olfactoria may have been severed at the level of the cribriform plate. The histopathologic changes in the olfactory receptors that were revealed by electron microscopy may have resulted from the inability of regenerating axons to reach their normal site of synaptic contact--the second-order neurons (mitral cells) in the olfactory bulb of the brain.

Electron microscopy of human olfactory epithelium reveals a new cell type: the microvillar cell.

The olfactory epithelium of mammals is generally considered to consist of 3 cell types: basal cells, supporting (sustentacular) cells, and ciliated olfactory receptors. We have completed a detailed ultrastructural study of the fine structure of the human olfactory mucosa. In our electron microscopic observations of biopsies of human olfactory epithelium taken from normal, consenting volunteers under local anesthesia, we have consistently observed a fourth cell type, the microvillar cell, located near the epithelial surface. The apical end of these flask-shaped, electron-lucent cells gives rise to a tuft of microvilli that project into the mucus layer lining the nasal cavity. The cell body itself contains bundles of microfilaments, mitochondria, a well-developed smooth endoplasmic reticulum, a prominent Golgi complex, electron-dense vesicles that resemble lipofuscin granules, free ribosomes, and occasional cisternae of the rough endoplasmic reticulum. A thin, axon-like cytoplasmic process extends from the basal pole of the cell and travels through the epithelium toward the lamina propria. Although there is no physiological evidence that bears upon the function of the microvillar cell, its ultrastructure suggests it may be a bipolar sensory neuron. Based upon morphological and phylogenetic considerations, the authors speculate the microvillar cell represents a second morphologically distinct class of chemoreceptor in the human olfactory mucosa.

The fine structure of the olfactory mucosa in man.

This report gives a detailed description of the fine structure of the olfactory mucosa in man. Using a special biopsy instrument and technique, fresh biopsies of olfactory epithelium were taken under local anaesthesia from eight normal volunteers. Transmission electron microscopy reveals that human olfactory epithelium has four major cell types: ciliated olfactory receptors, supporting cells, basal cells and microvillar cells. The ciliated olfactory receptors, as in other mammals, are bipolar neurons; the dendrite tip, modified to form the olfactory vesicle, bears 10-30 cilia that lack dynein arms. The supporting cells, markedly different from the goblet cells of respiratory epithelium, are not specialized for mucus secretion. Instead they are equipped to contribute materials to, and remove materials from, the surface mucus. The basal cells are stem cells that serve to replace epithelial cells and receptors lost during normal turnover or injury. In addition to ciliated olfactory neurons, supporting cells and basal cells, the human olfactory mucosa contains a distinct fourth cell type, the microvillar cell, of unknown function. The apical pole of the cell sends a tuft of short microvilli into the nasal cavity; its basal pole gives rise to a slender cytoplasmic process that resembles an axon. If microvillar cells prove to be sensory cells, the current concept of the human olfactory epithelium will have to be revised to include two morphologically distinct classes of receptors.

Suppression of reflex postural tonus: a role of peripheral inhibition in insects.

Postural reflexes act through a single excitatory motoneuron of the several that innervate a flexor muscle of the cockroach leg. A peripheral inhibitory neuron whose axon accompanies this excitatory motoneuron is able to suppress muscle tensions developed from postural reflexes without affecting centrally generated muscle tensions. The inhibitory neuron could thus serve to rapidly suppress postural tensions at the initiation of escape.

Biopsy of human olfactory mucosa. An instrument and a technique.

We have developed an instrument that obtains small biopsy specimens of human olfactory mucosa suitable for electron microscopy. Following local anesthesia of the patient, the instrument is introduced through the naris and advanced 60 to 70 mm between the septum and middle turbinate. The cutting edge is pressed against the olfactory area and then is withdrawn with the mucosal specimen. This procedure has been performed successfully on 12 people with no substantial adverse effects. We have provided the indications for the use of this technique and its possible applications.

Calcium-binding sites on sensory processes in vertebrate hair cells.

Vertebrate lateral line and vestibular systems center their function on highly mechanosensitive hair cells. Each hair cell is equipped with one kinocilium (which resembles a motile cilium) and 50-100 actin-containing stereocilia (which resemble microvilli) at the site of stimulus reception. This report describes electron-microscopic localization of calcium-binding sites on the sensory processes of vertebrate hair cells. Using the Oschman-Wall technique for calcium localization [Oschman, J. L. & Wall, B. J. (1972) J. Cell Biol. 55, 58-73] together with electron-probe x-ray microanalysis of thin sections, we observed: (i) calcium- and iron-containing deposits in the region of the ciliary necklace in goldfish lateral line hair cells, (ii) calcium deposits upon the surface of stereocilia of hair cells of the bullfrog inner ear, and (iii) calcium deposits upon stereocilia of hair cells of the guinea pig vestibular system.

A somatotopic organization of groups of afferents in insect peripheral nerves.

Sensory axon projections in the main leg nerves of two orthopteran insects, the cockroach Periplaneta americana and the grasshopper Melanoplus bivittatus, were studied by observing patterns of axonal degeneration after ablation of different leg segments. The patterns of degeneration seen in transverse sections of the leg nerves, close to their entrance to the central nervous system (thoracic ganglion) show that sensory axons occur in constant positions in the leg nerves. When distal leg segments are removed, a discrete area of degeneration is found in the leg nerve along its posterior edge. More proximal ablations produce larger areas of degeneration that progressively extend into the anterior half of the nerve. A comparison of the patterns of degeneration produced by different leg ablations shows a posterior to anterior laminar arrangement of groups of sensory axons that corresponds to a distal to proximal map of the leg. This mapping has been confirmed by localized ablations of small groups of leg sensory receptors.