Biomedical subjects

T V Getchell

Publications and source records attributed to T V Getchell.

At least 73 records · Page 4Linked to original sources

PubMed1988

Localization of substance P and neurofilament immunoreactive fibers in the lumbar facet joint capsule and supraspinous ligament of the rabbit.

An indirect immunofluorescence method was utilized to identify substance P-like immunoreactive (SPLI) and neurofilament protein immunoreactive (NFIR) fibers in the lumbar facet joint capsule and supraspinous ligament of the rabbit. The results demonstrated a large population of NFIR fibers, indicating that these tissues are richly innervated, and a smaller population of SPLI fibers. In some fibers, neurofilament protein and substance P were colocalized. The data suggest that the facet joint capsule and the supraspinous ligament contain SPLI nociceptive fibers that could be a source of low back pain.

Animals↗

PubMed1988

Ion transport across the frog olfactory mucosa: the action of cyclic nucleotides on the basal and odorant-stimulated states.

The action of cyclic nucleotides on the short-circuit current across the isolated bullfrog olfactory mucosa was studied both in the absence and presence of odorants. 8-Bromo-cAMP applied to the ciliated side of the mucosa caused a concentration-dependent, reversible increase in the basal short-circuit current, but not when it was applied to the submucosal side. The current had a sigmoidal concentration dependence described by the Hill equation. The magnitude of the odorant-evoked current was enhanced after bathing the ciliated side with cAMP analogs or modulators of intracellular cAMP. GTP gamma S added to the ciliated side increased the odorant-evoked current, while GDP beta S caused a decrease. Current transients induced by stimulating the ciliated side with either pulses of odorant or 8-bromo-cAMP were partially suppressed by amiloride, but only when amiloride and stimulant were presented simultaneously. Pulses of 8-bromo-cAMP and odorant presented simultaneously resulted in currents that added nonlinearly. In the absence of odorant, 8-bromo-cGMP caused a concentration-dependent decrease in net inward current that was reversed by 8-bromo-cAMP. Odorant-evoked currents were also reduced by 8-bromo-cGMP, and these could not be reversed by 8-bromo-cAMP. The results indicate that one type of olfactory transduction process involves the activation by cAMP of an inward current through an amiloride-sensitive apical ion channel and that this mechanism is mediated by a stimulatory G-protein.

8-Bromo Cyclic Adenosine Monophosphate↗

PubMed1988

Induction of odorant-evoked current transients in Xenopus oocytes injected with mRNA isolated from the olfactory mucosa of Rana pipiens.

Total mRNA isolated from the olfactory mucosa of the frog, Rana pipiens, and from rat pup brains was microinjected into mature oocytes isolated from Xenopus laevis. Both populations of voltage-clamped oocytes as well as controls were non-responsive to an odorant mixture and other ligands two days later. On the 3rd day, oocytes microinjected with rat brain mRNA responded to 5-hydroxytryptamine but not to an odorant mixture, while oocytes injected with Rana pipiens olfactory mucosal mRNA responded to an odorant mixture but not to other ligands. The magnitude of the small-amplitude, long-duration inward current transients recorded in response to odorant stimulation was enhanced by preincubating the oocytes with 8-bromo cAMP. The data suggest that odorant-gated ion channels associated with sensory transduction in the primary olfactory system were translated and inserted into the oocyte membrane. The results also indicate that the Xenopus oocyte system may be a useful preparation in which to investigate further the expression of membrane events associated with olfactory transduction.

8-Bromo Cyclic Adenosine Monophosphate↗

PubMed1988

Ultrastructural characteristics of sustentacular cells in control and odorant-treated olfactory mucosae of the salamander.

The ultrastructural characteristics of five morphologically distinct regions of sustentacular cells in the salamander olfactory mucosa are described. 1) The apical region was characterized by a microvillar surface that lay below the level of the olfactory knob of olfactory receptor neurons and contained endosome-like vesicles and a filamentous array at the level of the zonula adherens. 2) The supranuclear region contained rough and smooth endoplasmic reticulum, a Golgi complex, and secretory vesicles. Few sustentacular cells showed morphological signs of secretion, suggesting a low rate of baseline secretory activity. 3) The nuclear region contained the cylindrical nucleus surrounded by a thin band of cytoplasm containing bundles of filaments. 4) The central stalk contained filamentous arrays, Golgi-like cisternae, multivesicular bodies, and peroxisomes. Cytoplasmic veils that extended from the central stalk contained filamentous aggregates. 5) The basilar expansion had a complex series of lateral and basal folds. The lateral folds enveloped extracellular material and nonmyelinated axons of the receptor neurons. The basal folds formed complex interdigitations with the basal lamina, particularly in regions occupied by blood vessels and the acini of Bowman's glands in the subjacent lamina propria. These characteristics, and the presence of endosome-like vesicles and mitochondria, suggest that the basilar expansion is metabolically active and participates in cellular transport of material. Treatment with the odorant 2-isobutyl-3-methoxypyrazine caused ultrastructural changes in the apical and supranuclear regions that were associated with secretion and in the basilar expansion region that were indicative of an increase in metabolic and transport activity.

Animals↗

PubMed1987

Spectrophotometric determination of cation concentrations in olfactory mucus.

Spectrophotometric techniques were used to determine the concentrations of Na+, K+ and Ca2+ in the olfactory mucus of frogs. The mean concentrations in mEq/l were: [Na+], 52.7 +/- 4.1; [K+], 10.6 +/- 1.9 and [Ca2+], 10.7 +/- 1.7. Topical application of the odorant cineole was associated with statistically significant increases in [Na+] and [Ca2+]; the secretagogues methacholine and isoproterenol induced transient increases in [Na+]. Cineole and methacholine caused sustained increases in [Na+]/[K+] from the control value of 5:1, while isoproterenol caused a transient increase followed by a decline. The results indicate that the cation concentrations in olfactory mucus samples are more similar to those derived from secretory tissue than to those found in the extracellular fluids surrounding typical neural tissue.

Animals↗

PubMed1987

Ion transport across the frog olfactory mucosa: the basal and odorant-stimulated states.

The Ussing method was adapted to study the basal electrolyte transfer as well as the events that occur upon odorant stimulation in frog olfactory mucosa. The unstimulated short-circuit current was due mainly to a furosemide-sensitive ion transport system on the apical side of the olfactory mucosa. This current was not amiloride sensitive. The current-voltage relationship of the unstimulated state was linear. That of the odorant-evoked current was non-linear and amiloride-sensitive. Ouabain caused collapse of both the unstimulated and odorant-stimulated short-circuit current. In this case, voltage-clamping the tissue to non-zero values restored the odorant-evoked current with polarity depending on that of the clamping voltage. This suggested that the direction of the current is determined by that of the sodium electrochemical potential difference. Our results indicate that the unstimulated short-circuit current occurs through an apical sodium cotransport system, while the odorant-evoked current is due to odorant-activated, passive sodium channels that are amiloride sensitive.

Amiloride↗

PubMed1987

Odorant stimulation of secretory and neural processes in the salamander olfactory mucosa.

Topical application of the odorants guaiacol (10(-3) mol/l, 1-30 min) and 2-isobutyl-3-methoxypyrazine (IBMP, 10(-5)-10(-3) mol/l, 15 min) caused time- and concentration-dependent reductions in the secretory granule content of acinar cells of the superficial Bowman's glands (sBG) and moderate to extensive vacuolation in acinar cells of sBG and deep olfactory glands (dG). Topical application of 9.8 mg/ml scopolamine 10 min before 10(-4) mol/l IBMP significantly reduced the amount of secretory granule depletion from sBG compared to that seen with IBMP alone and resulted in less extensive vacuolation in sBG and dG acinar cells. The i.p. injection of 42 mg/kg propranolol 10 min before topical application of 10(-4) mol/l IBMP had no effect on the action of IBMP. Guaiacol and IBMP also had time- and concentration-dependent effects on the secretory activity of sustentacular cells in the olfactory epithelium. The protrusion of secretory material into the mucociliary matrix that covers the epithelial surface and vacuolation within the secretory material resulted from odorant application. Scopolamine and propranolol had no effects on the action of IBMP on sustentacular cell secretory activity. When applied in the vapor phase, guaiacol elicited action potentials recorded from individual olfactory receptor neurons; the impulse frequency was concentration-dependent and showed tonic and phasic components when the duration of stimulation was varied. Low to moderate concentrations of IBMP delivered in the vapor phase evoked monophasic negative slow voltage transients recorded from the surface of the olfactory mucosa. The amplitudes of these transients increased with increasing stimulus concentrations. Higher concentrations or longer stimulus durations evoked longer-latency positive-voltage generating processes and negative afterpotentials. The properties of the electrophysiological responses to both odorants were characteristic of responses evoked by a wide variety of 'typical' odorants.

Animals↗

PubMed1986

Amphibian terminal nerve: distribution revealed by LHRH and AChE markers.

Immunocytochemical and histochemical studies in the tiger salamander and bullfrog demonstrated the presence of luteinizing hormone-releasing hormone-like immunoreactive (LHRH-ir) material and acetylcholinesterase (AChE) in the terminal nerve (TN). Immunoreactive perikarya and processes were found within the olfactory, vomeronasal and trigeminal nerves and in the nasal epithelium. Central TN projections consisted of fibers terminating in the olfactory bulb and bundles that projected to another group of LHRH-ir perikarya in the preoptic region. Up to 4 weeks following hypophysectomy, the labeling intensity and number of TN-immunoreactive neurons were not altered. Acetylcholinesterase histochemistry in the salamander revealed two distinct groups of neurons associated with the TN: a lightly labeled group of fusiform perikarya was located in the olfactory nerve proper and a more heavily labeled group of larger oval perikarya was found within AChE-positive trigeminal fascicles in the ventral mucosa. This study has demonstrated that the amphibian TN follows olfactory, vomeronasal and trigeminal nerves to reach peripheral targets in the nasal mucosa. The projection of TN fibers to discrete olfactory bulb glomeruli, especially evident in the bullfrog, suggests that the TN functions in odor processing. The TN projection to the preoptic region in both of these amphibians implicates the TN in reproductive processes.

Acetylcholinesterase↗

PubMed1986

Functional properties of vertebrate olfactory receptor neurons.

The interaction of an odorant with the chemosensitive membrane of olfactory receptor neurons initiates a sequence of molecular and membrane events leading to sensory transduction, impulse initiation, and the transmission of sensory information to the brain. The main steps in this sequence are summarized in Figure 6. Several lines of evidence support the hypothesis that the initial molecular events and subsequent stages of transduction are mediated by odorant receptor sites and associated ion channels located in the membrane of the cilia and apical dendritic knob of the olfactory receptor neuron. Similarly, the membrane events associated with impulse initiation and propagation are mediated by voltage-gated channels located in the initial axonal segment and the axolemma. The ionic and electrical events associated with the proposed sequence have been characterized in general using a variety of experimental techniques. The identification, localization, and sequence of membrane events are consistent with the neurophysiological properties observed in specific regions of the bipolar receptor neuron. The influence of other cells in the primary olfactory pathway such as the sustentacular cells in the olfactory epithelium, the Schwann cells in the olfactory nerve, and the astrocytes in the olfactory nerve layer in the olfactory bulb on the physiological activity of the olfactory receptor neuron is an emerging area of research interests. The general principles derived from the experimental results described in this review provide only a framework that is both incomplete and of necessity somewhat speculative. As noted in the Introduction, the multidisciplinary study of the primary olfactory pathway is undergoing a renaissance of research interest. The application of modern biophysical, cell, and molecular biological techniques to the basic issues of odorant recognition and membrane excitability will clarify the speculations and lead to the establishment of new hypotheses. Three broad areas of research will benefit from such studies. First, the application of biophysical techniques will lead to a detailed characterization of the membrane properties and associated ion conductance mechanisms. Second, the isolation and biochemical characterization of intrinsic membrane and cytosolic proteins associated with odorant recognition, sensory transduction, and the subsequent electrical events will result from the utilization of cell and molecular biological techniques.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗

PubMed1984

Histological and histochemical studies of the secretory components of the salamander olfactory mucosa: effects of isoproterenol and olfactory nerve section.

Secretory components of the salamander olfactory mucosa, sustentacular cells (SC), and Bowman's glands (BG), were examined histologically and histochemically. In the aquatic larval salamander, SC in sensory grooves contained secretory granules; the submucosa contained a single layer of homogeneous, ductless glands. In the land-dwelling adult salamander, SC spanning a flat epithelial sheet contained vesicles. Subjacent to the epithelium in both dorsal and ventral mucosae lay BG whose ducts opened at the surface of the epithelium. In the ventral mucosa, two additional layers of olfactory glands (OG) lying below the BG were identified; ducts were not observed in association with the OG. The beta-adrenergic agonist isoproterenol caused depletion of secretory granules from BG and OG of larval, young, and adult salamanders but had no discernible effect on SC. Histochemical techniques (Alcian blue at pH 2.5 and pH 1.0, high-iron diamine, and the periodic acid-Schiff reaction) demonstrated that SC contained neutral, acidic, and small amounts of sulfated mucopolysaccharides (MPS), BG and OG contained only neutral MPS. In contrast, glands under adjacent respiratory epithelium contained both acidic and sulfated MPS. Unilateral olfactory nerve section ( ONX ) caused changes in the histochemical reactivity of acidic and sulfated MPS in SC on the ipsilateral and later on the contralateral side. Neutral MPS staining became enhanced first in the OG that lay under the BG, then in BG cells, and later in the deepest OG layer. Ipsilateral changes preceded contralateral ones. At 24 days post- ONX , some acinar cells in the deep OG contained acidic but not sulfated MPS.

Animals↗

PubMed1984

Perireceptor and receptor events in vertebrate olfaction.

In this article we have summarized the basic information which identifies several key issues in the study of perireceptor and receptor events in vertebrate olfaction. We have emphasized the biophysical and biochemical data which have established a pivotal role for the olfactory mucus in the access of odorants to receptor sites as well as their clearance from the micro-environment. In addition, based on initial reports in the literature, we have postulated that the uptake of odorants by cells in the olfactory epithelium and their subsequent enzymatic degradation is an important mechanism in odorant removal. Hence, the pre- and post-interactive events in vertebrate olfaction play a key role in molecular recognition, sensory transduction and receptor desensitization. Study of the primary events in vertebrate olfaction is an increasingly active area of research in neurobiology. Application of contemporary techniques in cell and molecular biology as well as biochemistry and cellular biophysics is yielding new insights into the process and into establishing new hypotheses to be tested.

Animals↗

PubMed1983

Morphological relations between the receptor neurons, sustentacular cells and Schwann cells in the olfactory mucosa of the salamander.

The morphological characteristics of olfactory receptor neurons and their relations to the sustentacular cells and to the sheath cells of Schwann in the olfactory mucosa of the salamander (Ambystoma maculatum et tigrinum) were studied using a modification of the rapid Golgi technique. The bipolar receptor neurons had a fusiform-shaped cell body whose apical pole gave rise to a surface-reaching dendrite and a basal pole which gave rise to an axon. The length and width of the dendrite, although variable, were positively correlated with the relative depth at which the cell body was located in the sensory epithelium. Beyond the initial segment, the axon had a sinuous course prior to its entrance into the lamina propria. Within the lamina propria, the axons were associated with the sheath cells of Schwann to form the olfactory nerve fascicles. The processes of adjacent sheath cells formed an elaborate network of continuous cavities through which the axons coursed. Two morphologically distinct varieties of sustentacular cells, designated types I and II, were found in the sensory epithelium. Both types had a columnar profile consisting of an elongated cell body, a central stalk, and a basilar expansion of the stalk found at the junction of the epithelium with the lamina propria. The central stalk of type I sustentacular cells was unbranched, whereas that of type II cells gave rise to riblike processes from which cytoplasmic veils extended to envelop the cell bodies of receptor neurons. The basilar expansions were found in close apposition to the wall of capillaries or to acinar cells of Bowman's glands located in the most superficial region of the lamina propria. The morphological relationships and possible interdependencies among receptor neurons, the types of sustentacular cells, and the sheath cells are discussed.

Animals↗

PubMed1981

Ultrastructural changes in olfactory receptor neurons following olfactory nerve section.

Unilateral olfactory nerve section was performed in the salamander, Ambystoma tigrinum. An ultrastructural study was performed to investigate the changes occurring during degeneration and replacement of the mature olfactory receptor neurons. Experimental and contralateral control tissues were examined following postoperative survival periods ranging from 12 hours to 90 days. Normal bipolar receptor neurons have a fusiform cell body containing a thin rim of cytoplasm and an ovoid nucleus with a characteristic "checkerboard" chromatin pattern. A single apical dendrite projects to the surface of the epithelium, where numerous cilia extend from its apex into the overlying mucus. A single, unmyelinated, unbranching axon originates at the basal pole of the cell. After nerve section, retrograde degeneration of the mature neurons occurs. Early degenerative changes include pronounced condensation of the nuclear chromatin, increased number of nuclear membrane infoldings, and dilation of the space between the membranes of the nuclear envelope. At a later stage, the cytoplasm of the cell increases in volume and its organelle systems break down, resulting in accumulation of various forms of cell inclusions. Subsequently, proliferation of cells in the basal region of the epithelium occurs. Between 3 week and 2 months following nerve section, these cells differentiate into mature neurons. By 3 months, neurons within the epithelium have resumed their normal ultrastructure. Correlation of the time course of the ultrastructural changes with previously reported neurophysiological studies indicates that neuronal activity of the epithelium is dependent upon the presence of fully differentiated olfactory receptor neurons.

Ambystoma↗

PubMed1981

Neurogenesis in olfactory epithelium: loss and recovery of transepithelial voltage transients following olfactory nerve section.

1. Unilateral olfactory nerve section was performed on the salamander, Ambystoma tigrinum. Physiological recordings and macroscopic observations were made to investigate the physiological correlates of functional recovery in the olfactory epithelium. 2. Slow transepithelial voltage transients, Veog, evoked by several odorous stimuli systematically decreased in amplitude during the initial 7 days and were not recorded at 10 days following nerve section, suggesting retrograde degeneration of receptor neurons. This was true for negative Veog(-), and positive, Veog(+), response components. Responses obtained from the untreated contralateral side of each animal remained similar to nonaxotomized controls. 3. Progressive recovery of the voltage transients was studied at 24, 45, 80, and 100 days following nerve section. At all stages of recovery, the wave form and time course of the responses were characteristic for each stimulus. This suggested that the response properties of the newly differentiated neuronal population were similar to those of the mature population. 4. At 100 days, response amplitudes evoked by all stimuli were similar to control values at all recording sites on the epithelial surface. The simultaneous loss and recovery of positive and negative components of the Veog indicated that the sources of both are dependent on the presence of functionally mature olfactory receptor neurons. 5. Visual inspection indicated that the olfactory nerve was reconstituted and reconnected to the olfactory bulb between 30-60 days following transection. The fact that physiological activity was recorded in the epithelium prior to this event suggests that molecular recognition and sensory transduction are not dependent on connectivity with the olfactory bulb. 6. It is concluded that physiological recovery of the olfactory receptor cell population occurs following axotomy. The time course of recovery was consistent with morphological evidence (see Ref. 57), indicating that newly differentiated receptor neurons are derived from cells in the basal region of the epithelium and replace the population lost through retrograde degeneration.

Ambystoma↗

PubMed1981

Physiological activity of newly differentiated olfactory receptor neurons correlated with morphological recovery from olfactory nerve section in the salamander.

1. Extracellular unitary recordings were made from the olfactory epithelium of the salamander, Ambystoma tigrinum, at numerous time points following olfactory nerve section. Unitary response properties were correlated with histological examination of the same tissues. 2. At 10 days following nerve section, unitary activity was rarely recorded in all regions of the epithelium. Histological examination indicated that virtually the entire mature olfactory receptor cell population had undergone retrograde degeneration. Transneuronal degeneration was not observed in the olfactory bulb, although the olfactory nerve and glomerular layers were substantially reduced in size. 3. At subsequent times, unitary impulse activity gradually returned, consisting of both spontaneous activity and odor-evoked discharges. Anatomical recovery of the olfactory epithelium preceded that of the olfactory bulb. A positive correlation was found between neuronal differentiation in the olfactory epithelium and the recovery of receptor cell function. 4. Patterns of spontaneous activity, odor specificities, intensity-response functions, and adaptive properties studied in newly differentiated olfactory receptor neurons were indistinguishable from those observed in control units. This indicated that these properties were intrinsic to the receptor neurons. 5. Spontaneously active and responsive units were encountered prior to olfactory nerve connection with the bulb. It is concluded that receptor neurons pass through two phases of functional maturity: the first independent of bulbar contact and the second dependent on presumed synaptic contact with bulbar neurons.

Ambystoma↗

PubMed1980

The location of olfactory receptor sites. Inferences from latency measurements.

Excitatory responses recorded from vertebrate olfactory sensory neurons are characterized by long latencies compared with those from other sensory receptors. Explanations which assume free access of the stimuli to receptor molecules presumably located on the olfactory cilia necessarily imply an intrinsic delay in the transduction mechanism. In contrast, the possibility of restricted or delayed access due to diffusion of the stimulus to molecular receptors located on the dendritic know or proximal portions of the cilia suggests transduction processes having time courses similar to those in other sensory systems. We show that the threshold stimulus concentrations and the latency of the excitatory response of the salamander can be predicted primarily on the basis of a diffusional delay and that the receptor molecules are well below the surface of the mucus. Examination of response latencies for other species reported in the literature support the generality of diffusional delay. The predicted location of molecular receptor sites is largely insensitive to assumptions based on the mode of clearance of the stimuli. Additional access restrictions are discussed but are shown to generate qualitatively different latency functions than does diffusion, suggesting that they exert only minor influences on latency and threshold characteristics.

Action Potentials↗

PubMed1978

Denervation of the primary olfactory pathway in mice. V. Long-term effect of intranasal ZnSO4 irrigation on behavior, biochemistry and morphology.

Intranasal irrigation of mice with 0.17 M ZnSO4 solution results in the immediate and total loss of the ability to find a buried food pellet. This anosmia persists for 6 weeks in at least 80% of the treated mice and for 4 months in half of the animals. This marked behavioral effect is matched by a long-term reduction of the levels of carnosine synthesis and transport in the primary olfactory pathway. These biochemical parameters are virtually undetectable at two weeks after treatment and even at one year after treatment do not exceed 5-10% of average control values. Light microscopic observations of tissues of the primary olfactory pathway at various times after treatment are consistent with these observations and indicate a substantial destruction of the olfactory epithelium with subsequent atrophy of the olfactory bulb. At very long intervals after treatment, some receptor regeneration is apparent with accompanying reinnervation of the olfactory bulb. Estimates from microscopy and biochemistry suggest that much less than 10% of the normal complement of functioning receptor cells is adequate to give apparently normal food-finding behavior.

Alanine↗

PubMed1978

Responses of olfactory receptor cells to step pulses of odour at different concentrations in the salamander.

1. The response properties of single olfactory receptor cells in the salamander have been analysed in unitary recordings obtained with platinum-black metal-filled micro-electrodes. 2. Stimulation has been carried out using an apparatus which delivers odour pulses of abrupt onset, steady plateau and abrupt termination. The pulses have been monitored near the site of stimulation on the olfactory epithelium during the experiments. 3. The main type of response was a discharge of impulses that was time locked to the stimulus pulse. The pattern of the responses consisted of a relatively brief latency of onset, a rapid rise in impulse frequency, a continuation of impulse firing during the plateau of the pulse, and an abrupt termination of the discharge correlated with the termination of the pulse. 4. There was a clear relationship between the receptor responses and odour concentration. In general, impulse firing frequency increased with increasing odour concentration. The firing frequency ranged from approximately 1--3 impulses/sec at threshold, up to 20 impulses/sec at the highest concentration. 5. Two types of reduced impulse activity were observed. One occurred after the termination of the pulse and lasted 1--3 sec; this was a common occurrence. The other type was seen during a pulse as a reduction of impulse activity compared to the background level; this type was rarely observed. 6. The receptor responses resembled those of mitral cells in the olfactory bulb to odour pulses in their sensitivity to odour concentration. They differed in that mitral cells show primary response categories consisting of brief excitation followed by suppression, and pure suppression, that are rarely seen at the receptor level. These differences may be ascribed to synaptic interactions in the olfactory bulb. 7. It is concluded that the majority of receptor cells have a stereotyped discharge response pattern and a systematic relation to odour concentration. These properties appear to reflect the simple time course of the odour pulses used in these experiments. This represents an initial step toward analysing olfactory coding at the receptor level using stimuli controlled in a manner similar to that used in other sensory systems.

Action Potentials↗