Biomedical subjects

T V Getchell

Publications and source records attributed to T V Getchell.

At least 55 records · Page 3Linked to original sources

PubMed1992

Fine structural aspects of secretion and extrinsic innervation in the olfactory mucosa.

The mucus at the surface of the olfactory mucosa constitutes the milieu in which perireceptor events associated with olfactory transduction occur. In this review, the ultrastructure of olfactory mucus and of the secretory cells that synthesize and secrete olfactory mucus in the vertebrate olfactory mucosa is described. Bowman's glands are present in the olfactory mucosa of all vertebrates except fish. They consist of acini, which may contain mucous or serous cells or both, and ducts that traverse the olfactory epithelium to deliver secretions to the epithelial surface. Sustentacular cells are present in the olfactory epithelium of all vertebrates. In fish, amphibia, reptiles, and birds, they are secretory; in mammals, they generally are considered to be "non-secretory," although they may participate in the regulation of the mucous composition through micropinocytotic secretion and uptake. Goblet cells occur in the olfactory epithelium of fish and secrete a mucous product. Secretion from Bowman's glands and vasomotor activity in the olfactory mucosa are regulated by neural elements extrinsic to the primary olfactory neurons. Nerve fibers described in early anatomical studies and characterized by immunohistochemical studies contain a variety of neuroactive peptides and have several targets within the olfactory mucosa. Ultrastructural studies of nerve terminals in the olfactory mucosa have demonstrated the presence of adrenergic, cholinergic and peptidergic input to glands, blood vessels, and melanocytes in the lamina propria and of peptidergic terminals in the olfactory epithelium. The neural origins of the extrinsic nerve fibers and terminals are the trigeminal, terminal, and autonomic systems.

Animals↗

PubMed1992

Ultrastructural localization of sialylated glycoconjugates in cells of the salamander olfactory mucosa using lectin cytochemistry.

An indirect gold-labeling method utilizing the lectin from Limax flavus was employed to characterize the subcellular distribution of sialic acid in glycoconjugates of the salamander olfactory mucosa. The highest density of lectin binding sites was in secretory vesicles of sustentacular cells. Significantly lower densities of lectin binding sites were found in secretory granules of acinar cells of both Bowman's and respiratory glands. Lectin binding in acinar cells of Bowman's glands was confined primarily to electron-lucent regions and membranes of secretory granules. In the olfactory mucus, the density of lectin binding sites was greater in the region of mucus closest to the nasal cavity than in that closest to the epithelial surface. At the epithelial surface, the density of lectin binding sites associated with olfactory cilia was 2.4-fold greater than that associated with microvilli of sustentacular cells or non-ciliary plasma membranes of olfactory receptor neurons, and 7.9-fold greater than non-microvillar sustentacular cell plasma membranes. Lectin binding sites were primarily associated with the glycocalyx of olfactory receptor cilia. The cilia on cells in the respiratory epithelium contained few lectin binding sites. Thus, sialylated glycoconjugates secreted by sustentacular cells are preferentially localized in the glycocalyx of the cilia of olfactory receptor neurons.

Ambystoma↗

PubMed1992

Glutathione and gamma-glutamyl transpeptidase are differentially distributed in the olfactory mucosa of rats.

Components of the gamma-glutamyl cycle, including thiols, glutathione (GSH) and gamma-glutamyl transpeptidase (gamma-GT), were localized in the nasal mucosae of rats using histochemical and immunohistochemical methods. In olfactory mucosa, thiols were widely distributed, with intense staining in the mucociliary complex (MC), basal cells, acinar cells of Bowman's glands (BG), and olfactory nerve bundles, and with moderate staining in olfactory receptor neurons (ORNs). GSH was localized in MC, BG acinar cells, nerve bundles and, to a lesser extent, in ORNs. gamma-GT immunoreactivity was restricted to the MC and to basolateral and apical membranes of BG acinar and duct cells. The basolateral membrane of BG acinar cells, located in close association with blood vessels and connective tissue, showed granule-like immunoreactivity. In respiratory mucosa, all three compounds were localized in the MC and acinar cells of respiratory glands (RG). In the MC, gamma-GT immunoreactivity was associated primarily with brush borders of ciliated cells. Granular immunoreactivity was also apparent in the supranuclear region of RG acinar cells. These results demonstrate that components of the gamma-glutamyl cycle are localized in olfactory and respiratory glands, and that they are secreted into the mucus, where they may mediate perireceptor events such as detoxification and/or solubilization of air-borne xenobiotics, toxicants and odorants.

Animals↗

PubMed1992

Differential distribution of gamma-glutamyl cycle molecules in the vomeronasal organ of rats.

Molecules related to the gamma-glutamyl cycle, including thiols, glutathione (GSH) and gamma-glutamyl transpeptidase (gamma-GT) were identified histochemically and immunohistochemically in the vomeronasal organ of neonatal and adult rats. Thiols and GSH were distributed in the mucomicrovillar complex (MMC), vomeronasal receptor neurons and acinar cells of vomeronasal glands (VNG). gamma-GT was localized in the MMC and in the VNG, where it was associated mainly with the luminal surface of the acinar cells and ducts. The VNO of the neonates exhibited higher staining intensities for all compounds than that of the adults. The data indicate that components of the gamma-glutamyl cycle are present in the VNO and that they are secreted into mucus, where they may be associated with perireceptor events including clearance of pheromones and detoxification of xenobiotics.

Aging↗

PubMed1992

Immunolocalization of two cytochrome P450 isozymes in rat nasal chemosensory tissue.

The NMa and NMb isoforms of cytochrome P450 enzymes are expressed in three nasal chemosensory organs: the olfactory, septal and vomeronasal mucosae. The NMa isoform is widely distributed throughout the nasal mucosa whereas the NMb isoform is present primarily in the chemosensory mucosae. The localization of cytochromes P450 demonstrates that sustentacular cells in the olfactory and septal epithelia, the mucus of the vomeronasal organ and the acinar cells of glands in the lamina propria of all three chemosensory systems engage in xenobiotic metabolism and participate in odorant/pheromone clearance, a perireceptor process associated with chemosensory transduction.

Animals↗

PubMed1992

The effects of controlled mechanical loading on group-II, III, and IV afferent units from the lumbar facet joint and surrounding tissue. An in vitro study.

An in vitro model was developed to investigate the responses of afferent units in the lumbar spine to controlled loading as measured by a load-cell. The neuronal discharge was recorded simultaneously with loading. Three types of neuronal responses were observed. The first type of response involved phasic-type mechanoreceptors, which responded to movement, regardless of direction or initial position. The response did not outlast the movement phase of loading. These units may serve as velocity detectors. The second type of response was seen in slowly adapting low-threshold mechanoreceptors, which tended to respond to loading in the 0.3 to 0.5-kilogram range with an immediate and sustained increase in the rate of firing. This type of response appears to be associated with the activation of low-threshold group-II and group-III fibers, which were located in muscles and tendons inserting into the facet joint. The third type of response involved slowly adapting high-threshold mechanoreceptors, which could not be activated until a threshold of three to five kilograms had been exceeded. It appears that this type of response is at least partially due to the activation of high-threshold group-III and group-IV capsular afferent units, which may signal noxious mechanical stimulation.

Adaptation, Physiological↗

PubMed1992

Characterization of the immune barrier in human olfactory mucosa.

Immunologic defense factors in the human olfactory mucosa were localized immunohistochemically. Olfactory epithelium was identified with an antiserum to olfactory marker protein, specific for olfactory receptor neurons. Constituents of the secretory immune system, including IgA, IgM, secretory component, and J chain, were localized in the acinar and duct cells of Bowman's glands and in the mucociliary complex. In addition, B lymphocytes in the lamina propria near Bowman's glands displayed immunoreactivity for IgA, IgM, and J chain. Immunostaining also localized other humoral factors. Immunoreactivity for IgG was present throughout the stroma and in B lymphocytes in the lamina propria. Antibody to IgD stained numerous B lymphocytes clustered below the basement membrane. Antibody to IgE stained similarly distributed cells; toluidine blue staining demonstrated that many were mast cells. In addition, antibodies to IgD and IgE stained occasional intraepithelial B lymphocytes or mast cells. Two antimicrobial proteins, lactoferrin and lysozyme, were localized in Bowman's glands and the mucociliary complex. Thus, the human olfactory mucosa, which provides a direct neural route for pathogens to the brain, is a site for synthesis and secretion of immune and other defense factors.

Adolescent↗

PubMed1991

Ultrastructural localization of Na+/K(+)-ATPase in rodent olfactory epithelium.

The olfactory epithelium is comprised of bipolar sensory neurons, sustentacular cells, and basal cells. The sensory neurons have apical knobs and cilia, which project into the olfactory mucus toward the nasal lumen, and represent presumptive sites of odorant binding. Ionic currents, measured across this epithelium in both the resting and odorant-stimulated states, are known to be sustained, at least in part, by active transport of sodium. Information identifying the cellular sites of ion transport in olfactory sensory epithelium will therefore aid in elucidating the ionic mechanisms associated with olfactory transduction. The membrane-bound enzyme Na+/K(+)-ATPase mediates active ion transport in many other cells and tissues. We have consequently employed the cytochemical technique reported by Ernst (J. Histochem. Cytochem., 20 (1972) 23-38, 1322) to identify possible sites of elevated Na+/K(+)-ATPase activity in olfactory epithelium. This procedure detects inorganic phosphate (Pi) released from an artificial substrate (nitrophenyl phosphate) by enzyme catalytic activity. In the presence of strontium ion. Pi is precipitated near regions of high enzymatic activity, then converted to a product visible in the electron microscope. Parallel control preparations were incubated in media (1) supplemented with the specific Na+/K(+)-ATPase inhibitor ouabain (to abolish formation of specific reaction product); (2) with substrate deleted (to demonstrate possible non-specific binding of Sr2+ and/or Pb2+); or (3) with the necessary cofactor K+ deleted. In tissues incubated for demonstration of Na+/K(+)-ATPase activity, reaction product was associated with apical knobs, cilia and dendrites of olfactory receptor neurons at the apical surface. In the more proximal region of the epithelium, reaction product was associated with cell bodies and axons of the sensory neurons, and with the lateral membranes of sustentacular cells. Reaction product was deposited intracellularly, compatible with the known mechanism of the Na+/K(+)-ATPase enzymatic reaction. In control specimens incubated with ouabain, with substrate deleted, or with K+ deleted, only a small quantity of non-specific precipitate was observed. These results are discussed with reference to the various sodium currents implicated in olfactory transduction and transepithelial transport.

Animals↗

PubMed1991

Identification of sugar residues in secretory glycoconjugates of olfactory mucosae using lectin histochemistry.

Lectin histochemistry at the light microscope level was used to determine the distribution of sugar residues in secretory cells of the olfactory mucosae of salamander, hamster, and mouse. Differences in sugar composition and distribution of glycoconjugates found in sustentacular cells and acinar cells of Bowman's glands of these three animals were characterized. Oligosaccharides in secretory products of sustentacular cells in salamander olfactory mucosa contained sialic acid, galactose (Gal), N-acetylgalactosamine (GalNAc), N-acetylglucosamine (GlcNAc), fucose, and mannose residues. Glycoconjugates of these cells lacked terminal galactosyl-beta-(1,3)N-acetylgalactose (Gal beta 1,3GalNAc) residues. The sequences Gal beta 1,3GalNAc, N-acetyllactosamine (Gal beta 1,4GlcNAc), and GalNAc were penultimate to sialic acid residues. Sustentacular cells of mouse and hamster did not appear to contain O-linked oligosaccharides but stained for mannose-containing N-linked oligosaccharides. Glycoconjugates of acinar and duct cells of Bowman's glands in the salamander, hamster, and mouse contained variable amounts of beta(1,4)GlcNAc residues, and terminal N-acetyllactosamine, Gal beta 1,3GalNAc, and GalNAc residues. In the salamander, glycoconjugates of acinar cells possessed terminal GlcNAc residues but were not sialylated, while those of hamster and mouse generally stained for sialic acid but did not possess terminal GlcNAc residues. Secretory products of a subpopulation of rodent acinar cells also contained penultimate Gal beta 1,3GalNAc residues. Staining for sialic acid, Gal, GalNAc, and GlcNAc in glycoconjugates of rodents was often limited to a sub-population of Bowman's glands. This was especially noticeable in the mouse.

Ambystoma↗

PubMed1991

Immunohistochemical localization of components of the immune barrier in the olfactory mucosae of salamanders and rats.

Immunohistochemical techniques were used to investigate the cellular distribution of components of the secretory immune system, including secretory immunoglobulin, secretory piece, and J chain, as well as other immunoglobulins and nonspecific defense factors in the olfactory mucosae of salamanders and rats. In the salamander, secretory immunoglobulin M, and J chain were localized in duct and acinar cells of Bowman's glands, in B lymphocytes, and in sustentacular cells in immature regions of the olfactory mucosa. Lactoferrin and lysozyme were also present in Bowman's glands, in sustentacular cells in immature regions of the olfactory mucosa, and in blood cells in the lamina propria. Olfactory nerve section resulted in the presence of increased numbers of secretory immunoglobulin-immunoreactive B lymphocytes and in an altered distribution of IgM, secretory piece, and lactoferrin. In the rat, secretory immunoglobulin A and J chain were localized in duct and acinar cells of Bowman's glands and in B lymphocytes in the lamina propria. Secretory piece could be demonstrated in Bowman's glands only in rats that had a prior viral infection. Other defense factors, localized in the lamina propria, included IgG in the connective tissue stroma and in B lymphocytes, IgD-immunoreactive B lymphocytes, and IgE-immunoreactive cells that were identified as mucosal mast cells. Lactoferrin and lysozyme were present in serous acinar cells of Bowman's glands and in blood cells. These results demonstrate that the olfactory mucosa is protected from pathogenic invasion by the secretory immune system as well as other immunoglobulins, lactoferrin, and lysozyme.

Ambystoma↗

PubMed1991

Vomeromodulin, a putative pheromone transporter: cloning, characterization, and cellular localization of a novel glycoprotein of lateral nasal gland.

Vomeromodulin, a novel glycoprotein of lateral nasal gland, has been cloned by differential hybridization from nasal/olfactory tissue. The 2.2-kb mRNA directs the in vitro synthesis of a 60-kDa primary translation product in reticulocyte lysates. Differential sensitivity to endoglycosidases indicates that vomeromodulin is posttranslationally modified in vivo by N-glycosylation to form a 70-kDa glycoprotein of the complex type. Immunocytochemical localization with two different antipeptide antisera demonstrates that vomeromodulin is abundant in the lateral nasal glands and is also present in the posterior septal and vomeronasal glands. Most striking is the observation that it is highly concentrated in the mucus of the vomeronasal organ of Jacobson but is not detectable in the mucus of the main olfactory neuroepithelium. Evaluation of mRNA and protein distribution by Northern and Western analyses, respectively, indicates that vomeromodulin is absent from 15 other tissues. The glandular and mucosal distribution of this glycoprotein implies a transport function that may be related to the mechanisms by which pheromones of low volatility gain access to their receptors in the vomeronasal organ. These observations support our hypothesis that vomeromodulin participates in perireceptor events that facilitate the process of pheromone access and detection.

Amino Acid Sequence↗

PubMed1990

Expression of catfish amino acid taste receptors in Xenopus oocytes.

We demonstrate that poly (A+)RNA isolated from catfish barbels directs the expression of functional amino acid taste receptors in the Xenopus oocyte. The activity of these receptors is monitored in ovo by the two electrode voltage clamp technique. Specific conductance changes recorded in response to amino acid stimulation are analogous to those recorded electrophysiologically from intact catfish barbels. These responses exhibit specificity, reproducibility, rapid onset and termination, and desensitization to repetitive stimulation. A functional assay system that encompasses the full complement of transduction events from the ligand-receptor interaction to subsequent conductance changes is necessary to identify molecular components responsible for these events. Our results demonstrate that the Xenopus oocyte can be used to characterize and identify clones coding for amino acid taste receptors analogous to its use in studying receptor molecules for other neuroactive compounds.

Amino Acids↗

PubMed1990

Mechanosensitive afferent units in the lumbar facet joint.

The purpose of this study was to characterize somatosensory units of the lumbar facet joint, which may play a central role in idiopathic low-back pain. A laminectomy was performed on the lumbar spine of adult male New Zealand White rabbits. Receptive fields of mechanosensitive afferent units were investigated in the lumbar facet joint and adjacent surrounding tissues, and electrophysiological recordings were obtained from filaments of the dorsal root. Twenty-four units were identified in the region of the facet joint: ten, in the capsule of the joint; twelve, in the border regions between capsule and muscle or tendon; and two, in the ligamentum flavum. Of these units, two had a conduction velocity that was slower than 2.5 meters per second (group IV), fifteen had a velocity ranging from 2.5 to twenty meters per second (group III), and seven had a velocity faster than twenty meters per second. Seven units had a von Frey threshold of more than 6.0 grams, thirteen had a threshold of less than 6.0 grams, and four were not examined. Seven units in the facet joint responded to movement of the joint. Fourteen other mechanosensitive units were found in the muscle, tendon, and interspinous ligament; seven had a conduction velocity of 2.5 to twenty meters per second, and seven had a velocity that was faster than twenty meters per second.

Animals↗

PubMed1989

Ultrastructural evidence for peptidergic innervation of the apical region of frog olfactory epithelium.

Ultrastructural examination of the region near the olfactory epithelial surface of leopard frogs revealed the presence of nerve terminals just proximal to the zonula adherens between adjacent sustentacular cells, and between sustentacular cells and olfactory receptor neurons. Terminal varicosities, located about 20 nm from sustentacular cell membranes, contained numerous large-diameter dense-cored vesicles, small-diameter agranular vesicles, and mitochondria. On the basis of ultrastructural characteristics, they are identified as peptidergic sensory terminals.

Animals↗

PubMed1989

Ultrastructural localization and identification of adrenergic and cholinergic nerve terminals in the olfactory mucosa.

Pharmacological and ultrastructural methods were used to demonstrate alpha-adrenergic regulation of secretory granule content of acinar cells of Bowman's glands and to localize and identify adrenergic and cholinergic axonal varicosities and terminals in the olfactory mucosa of the tiger salamander. The alpha-adrenergic agonist phenylephrine caused secretory granule depletion from Bowman's glands; the alpha-adrenergic antagonist phentolamine partially blocked this effect. These observations were quantified using light microscopic computer-assisted morphometric techniques. Both drugs caused morphological signs of electrolye/water transport. Adrenergic axonal varicosities were identified by the presence of small granular vesicles (SGVs, 45-60 nm in diameter) containing electron-dense material that was enhanced by 5-hydroxydopamine loading and chromaffin reaction fixation techniques. Throughout the lamina propria, small fascicles with axons containing SGVs as well as varicosities and terminals with SGVs were located adjacent to blood vessels, Bowman's gland acini, and melanocytes. Mean vesicle diameters at these sites were 54 +/- 7 nm, 50 +/- 9 nm, and 56 +/- 8 nm, respectively; varicosities were located approximately 0.1-1.0 microns from their presumed cellular targets. Axonal varicosities containing small agranular vesicles (AGVs, 65 +/- 8 nm in diameter), identified as cholinergic by their size and by the absence of electron-dense material after 5-hydroxydopamine loading and chromaffin reaction fixation, were located between adjacent acinar cells. In addition, adrenergic varicosities containing SGVs (56 +/- 6 nm in diameter) were found within 1 micron of blood vessels associated with Bowman's gland ducts and sustentacular cells near the base of the olfactory epithelium. These results characterize the ultrastructural basis for adrenergic and cholinergic regulation of vasomotor tone and secretion within the olfactory mucosa.

Adrenergic Fibers↗

PubMed1989

Sensory innervation of soft tissues of the lumbar spine in the rat.

The purpose of this study was to investigate neurophysiologically and anatomically the soft tissues of the dorsal compartment of the lumbar spine in order to understand better their possible role in low back pain. The focus was primarily on the lumbar facet joint and supraspinous ligament of the Sprague-Dawley rat. Microdissection of the dorsal ramus and electrophysiological and neuroanatomical studies of the dorsal ramus and its terminations in paravertebral tissue revealed that (a) there are mechanosensitive, slowly adapting fibers in the rat lumbar facet joint capsule; (b) there are slowly adapting, mechanosensitive units in the rat supraspinous ligaments that respond to tensile loading; both types of mechanosensitive units have high threshold; (c) mechanical stimulation of these tissues sometimes elicits afterdischarges lasting several minutes; (d) many extracellular recordings from the medial branch of the dorsal ramus appear to be reflex activity to mechanical stimulation; (e) silver impregnation of the rat joint capsule reveals individual axons, very few of which were encapsulated, suggesting that they terminate in free nerve endings; and (f) the nerves of the rat facet joint capsule contain 68, 160, and 200 kdalton polypeptide subunits of neurofilament protein (NFP). These results indicate that neurons of lumbar facet joint capsules and ligaments in the back are sensitive to mechanical strain and that the higher threshold neurons may serve a nociceptive (pain) function.

Animals↗

PubMed1989

Peptidergic regulation of secretory activity in amphibian olfactory mucosa: immunohistochemistry, neural stimulation, and pharmacology.

The role of substance P in the regulation of secretion from sustentacular cells, Bowman's glands and deep glands in the amphibian olfactory mucosa was investigated using immunohistochemical, electrophysiological, and pharmacological methods. Substance P-like immunoreactive varicose fibers extended through the olfactory epithelium, terminating at or near the surface. In addition, immunoreactive varicose fibers innervated Bowman's glands, deep glands, and blood vessels in the lamina propria. Innervation of Bowman's gland was sparse, with fibers terminating on basal acinar cell membranes; deep gland innervation was abundant, with fibers often extending between acinar cells almost to the lumen. Stimulation of the ophthalmic branch of the trigeminal nerve resulted in slow potentials recorded at the surface of the olfactory epithelium. When the olfactory mucosae from trigeminal-stimulated animals were examined histologically, morphological signs of secretory activity were observed, suggesting that substance P was released from the trigeminal nerve terminals. Topical application of 10(-5) to 10(-3) mol substance P resulted in morphological signs of secretion that were very similar to those seen as a result of trigeminal stimulation. Thus, substance P released from trigeminal fibers may modulate secretory activity within the olfactory mucosa.

Animals↗

PubMed1989

Expression of the mammalian Na+-independent L system amino acid transporter in Xenopus laevis oocytes.

System L is primarily responsible for the Na+-independent transport of neutral amino acids, those with bulky chains such as leucine, isoleucine, phenylalanine, etc., into mammalian cells. mRNA from rat kidney and human lymphoid cells, when microinjected into Xenopus laevis oocytes, induced expression of this transport system. The expressed transport exhibits characteristics similar to those reported for the System L amino acid transporter from a variety of mammalian cells. Injection of size-fractionated mRNA indicates that the System L transporter in both the rat kidney and human lymphoid cells is encoded by mRNA of about 3 to 4 kb.

Amino Acid Transport Systems↗