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3-O-methyl-D-glucose and beta-alanine: rapid axoplasmic transport of metabolically inert low molecular weight substances.

The dynamics of the axoplasmic transport of free beta-alanine and 3-O-methyl-D-glucose were quantitatively characterized in vivo in the pike olfactory nerve following intranasal application of the tritiated substances. Both compounds were found to be taken up by the epithelium but not metabolized to a significant extent. Axoplasmic transport showed the same dynamics as rapid transport of protein and free leucine in this nerve. No synthesis or transport of carnosine was detectable. The results are in agreement with the view that rapid axoplasmic transport is capable of conveying free, low molecular weight molecules in a non-specific manner.

3-O-Methylglucose↗

The development of the olfactory mucosa in the mouse: electron microscopy.

The development of the olfactory epithelium from the 10th day of gestation of postnatal life has been examined electron miscroscopically in the mouse. At 10 days' gestation the epithelium is already differentiated into dark and pale cells, the former representing embryonic stem cells and the latter the developing receptors. Axons are also visible at this stage. At 11 days the first signs of dendrite formation appear, and at 12 days spheroidal terminal swellings containing numerous microtubules are present at the apices of receptor dendrites. Centriole clusters also appear in the receptor cell bodies and dendrites. From the 12th to the 16th day of gestation a few cilia are formed on the receptor endings. Final steps in the maturation of differentiating receptors begin on the 17th day of gestation, when membranous organelles and lysosomes increase greatly in numbers. However, immature receptors can still be found in the base of the epithelium in postnatal life. Supporting cells are first recognizable on the 17th day of gestation, derived apparently from the remaining stem cells. At the same time differentiated basal cells and glands of Bowman begin to appear. In the early develoment of the olfactory nerve bundles the axons have large and varying diameters, but later on axonal sizes are progressively reduced and the adult size range is achieved at about 18 days of gestation. The significance of these findings is discussed.

Age Factors↗

Suppression of evoked potentials with continued ion transport during anoxia in turtle brain.

A key to turtle brain survival during anoxia is continued ion transport and avoidance of anoxic depolarization. Previous findings that ATP concentration remained constant during prolonged anoxia and calculations that ATP production decreased indicate that compensatory processes, other than consumption of energy stores or increased anaerobic glycolysis, must also contribute to ion homeostasis and brain survival. To determine whether preservation of ion transport is associated with changes in electrophysiology during loss of oxidative metabolism, the brains of pentobarbital sodium-anesthetized turtles were electrically stimulated 1) to provoke measurable increments in extracellular K+ activity (a degrees k) for determination of rates of K+ reaccumulation at the stimulus site and 2) to elicit polysynaptic extracellular field potentials (evoked potentials) recordable in the olfactory bulb. During anoxia, base-line a degrees k rose only a few millimolar, and rates of reaccumulation of K+, incremented by stimulation were slightly but not significantly slowed. In contrast, postsynaptic orthodromic responses of olfactory bulb granule cells were markedly depressed by anoxia. Monosynaptic responses of granule cells to antidromic stimulation of the lateral olfactory tract were less affected, and compound action potentials in the olfactory nerve were unchanged by anoxia. These data suggest that synaptic transmission in turtle brain, as in that of mammal, is highly dependent on oxidative metabolism and that the turtle brain may effectively conserve energy for ion transport during anoxia by depression of electrical activity.

Animals↗

Membrane and synaptic properties of mitral cells in slices of rat olfactory bulb.

We have investigated the membrane properties and excitatory synaptic transmission of mitral cells in a slice preparation of rat olfactory bulb. In response to intracellular injection of depolarizing current, most mitral cells showed several distinct membrane properties: (1) delayed onset of firing (suggesting the presence of a type of potassium A current); (2) subthreshold oscillation of the membrane potential; and (3) repetitive firing of clustered action potentials during prolonged threshold stimulation. Olfactory nerve (ON) stimulation evoked a long-lasting EPSP in most of the mitral cells. This long EPSP was completely blocked by combined application of NMDA and non-NMDA receptor antagonists (20 microM CNQX and 100 microM APV), confirming that glutamate is the neurotransmitter at the synapses from ON to mitral cells. The ON-evoked EPSP was preceded by a prespike, which was resistant to membrane potential hyperpolarization at the soma. This fast prepotential may be indicative of an active response in the primary dendritic tufts of the mitral cells. Stimulation of the lateral olfactory tract evoked an antidromic pulse followed by a short EPSP, which could also be elicited independently of an antidromic spike in the recorded cell. Since the asymmetrical synapses so far observed on the mitral cells are all form the ON, this antidromically evoked EPSP may reflect self-excitation of a mitral cell by glutamate released from its own dendrites by antidromic impulse invasion, or/and lateral excitation by neighboring invaded dendrites.

2-Amino-5-phosphonovalerate↗

Localization and regulation of low affinity nerve growth factor receptor expression in the rat olfactory system during development and regeneration.

Nerve growth factor (NGF), a classic neurotrophic factor, promotes neuronal survival, maintenance, regeneration and differentiation in the peripheral nervous system and parts of the central nervous system. NGF activity is mediated by cell surface bound receptors including the low affinity NGF receptor (LNGFr) which is expressed by some peripheral and central neurons and is present on peripheral nerve Schwann cells during development and regeneration. The olfactory system is a useful model for the study of the role of LNGFr in neuronal development and regeneration. The growth of olfactory axons into the brain begins in the embryo and continues through the first few postnatal weeks. In mature animals there is persistent turnover and generation of olfactory receptor neurons (ORNs) and continuous growth of new axons into the olfactory bulb. These new axons grow along the preexisting olfactory pathway. In the mature olfactory system, LNGFr has been observed in the glomerular layer of the olfactory bulb, the target of ORNs. However, neither the cellular localization nor the developmental expression of LNGFr has been characterized. Here, we tested the hypothesis that LNGFr expression is developmentally regulated in the olfactory nerve and is reinduced following injury to the mature olfactory nerve. LNGFr-immunoreactivity (IR) was first observed in the olfactory mucosa at embryonic day (E)13 and in the olfactory nerve at E14. LNGFr-IR increased in the nerve during embryonic development, began to decrease at around postnatal day (P)5 and was scarcely detectable in normal adults. The staining pattern suggests that LNGFr is located on the olfactory nerve Schwann cells. Streaks of LNGFr-IR were present in the adult olfactory nerve. We reasoned that these streaks might represent transient reexpression of LNGFr associated with normal olfactory neuron turnover and replacement. Consistent with this hypothesis, LNGFr was robustly reexpressed in the adult olfactory nerve following lesion of the olfactory epithelium. Starting late in development (E21) and in the adult, LNGFr-IR was also observed on fibers in deep layers of the olfactory bulb. LNGFr-IR was also observed in neurons of the nucleus of the diagonal band (NDB) in the basal forebrain. NDB is the sole source of cholinergic afferents of the olfactory bulb. Thus, we tested the hypothesis that LNGFr in the deep layers of the olfactory bulb is located on NDB axons by making lesions of NDB. Following the lesion, LNGFr-IR disappeared in the deep layers of the olfactory bulb but remained in the glomerular layer.(ABSTRACT TRUNCATED AT 400 WORDS)

Acetylcholinesterase↗

Computation of frequency-to-spatial transform by olfactory bulb glomeruli.

A physiological simulation of 2.5% of the input and inhibitory neurons and 25% of the primary mitral/tufted cells in a single mammalian olfactory bulb glomerulus was constructed. This physiological simulation used the integrate-and-fire paradigm with realistic activation curves and synaptic delays. The dendritic integration incorporated non-linear interactive effects of individual cell excitatory and inhibitory post-synaptic potentials (PSPs) from both axodendritic and dendro-dendritic synaptic contacts. Refractory periods for granule-cell inhibition of mitral/tufted cell activity lead to relatively fixed-frequency rhythmic activity in the glomerulus, independent of the input frequency from the olfactory nerve. Though the frequency of mitral/tufted cell firing in bulb was approximately independent of input frequency, the number of cells active in the glomerulus was a roughly-linear function of input frequency to the glomerulus, indicating the mechanism's ability to function as a frequency-to-spatial encoder.

Animals↗

[Olfactory acuity and improvement of olfaction after total laryngectomy].

OBJECTIVE: To test and evaluate the olfactory function of patient after total laryngectomy, seek to a practical method to ameliorate olfactory function and rise the qualitative character of these patients. METHODS: Using the T&T olfactory examination to evaluate the olfactory function of 60 cases. Four cases olfactory mucosae were observed by electron microscope. Observing relation among the remains olfaction, the time after operation and whether or not undergone the voice reconstruction. And analyse the reasons of the above hyposomnia. Using the closing-mouth and nasal out-word airflow maneuver (CNOAM) as the intervention in the patients of tracheoesophageal fistula voice reconstruction after total laryngectomy to observe the amelioration after intervention. RESULTS: It shows various degree of hyposmia and anosmia in the cases after total laryngectomy with or without tracheoesophageal fistula voice reconstruction, with significant deference (P < 0.01) compared to the normal person respectively. There are precisely correlation among the time after operation and whether or not undergone the voice reconstruction. The longer time leads to less remaining olfaction. The patients after total laryngectomy without tracheoesophageal fistula voice reconstruction have lost their olfaction thoroughly within 5 years. But for the patients after total laryngectomy with tracheoesophageal fistula voice reconstruction, they have a middle hyposmia within 5 years, with significant deference (P < 0.01) between the patients in 5 years and after 5 years. There were significant differences (P < 0.01) between the values of patients with and without tracheoesophageal fistula voice reconstruction. The ultrastructure of 4 cases of olfactory epithelium shows the apoptosis change. After the treatment of CNOAM, the remaining olfaction of most patients were improved, with significant deference (P < 0.01) compared to those before the treatment of CNOAM. CONCLUSIONS: The proceed hypofunction of olfaction may be influenced by the reform of respiratory air, the extinction of air velocity bypass the nasal cavity and the apoptosis of epithelial cells in the patients after total laryngectomy. But if we give an early intervention study such as tracheoesophageal fistula voice reconstruction and CNOAM, the olfactory function may be maintenance. During the intervention, the ending of olfactory nerves may be get uninterrupt stimulation. This may help the patients keep a better existing quality than those fail to accept the interventions.

Adult↗

Expression of neu and Neu differentiation factor in the olfactory mucosa of rat.

The growth and differentiation of olfactory sensory neurons are regulated tightly. We had shown previously, by immunohistochemistry, that transforming growth factor-alpha (TGF-alpha) and epidermal growth factor (EGF) receptor are present in the olfactory epithelium of untreated adult rats and that TGF-alpha is a potent mitogen of olfactory epithelium in vitro. Expression of EGF receptor and TGF-alpha was detected primarily in horizontal basal cells and supporting cells but rarely in globose basal cells, which suggested that EGF receptor is not a likely candidate for the mitotic regulator of sensory neurons. In order to expand the search for candidate regulators, we have now examined other members of the EGF family of receptors and ligands. By utilizing reverse transcriptase-polymerase chain reaction (RT-PCR) methodology, we have detected the messenger RNA encoding the protein of the neu gene (p185neu) and Neu differentiation factor (NDF) isoforms in the olfactory mucosa. Immunohistochemical localization of p185neu and NDF indicates expression of these proteins in the olfactory epithelium of adult rats in regions where globose basal cells and immature sensory neurons are found, as well as in the ensheathing cells of the olfactory nerve. The presence of neu and NDF transcripts in the olfactory tissue and the localization of their encoded polypeptides to proliferative regions of the epithelium suggest involvement of these gene products in the regulated proliferation/differntiation of the sensory neurons.

Animals↗

Immunohistochemical observations of dividing cells in olfactory epithelium using anti-BrdU antibody.

To examine the dividing cells in the olfactory epithelium, an experiment using a novel immunohistochemical technique with bromodeoxyuridine (BrdU) was performed. Tissue specimens were obtained from the olfactory epithelium of guinea pigs at days 7 and 21 after olfactory nerve axotomy. BrdU uptake was detected in the epithelial cell layer directly above the basal cell layer rather than in the basal cells per se. The BrdU-immunoreactive cells were found more numerously at 7 days than at 21 days after axotomy. The basal cells showed no immunoreaction to the anti-BrdU antibody on either day. The cells reacting with the anti-BrdU antibody also showed no reaction to the anti-cytokeratin antibody used to identify the basal cells. These findings indicate that the cells showing mitotic activity have characteristics different from those of basal cells, which has been considered previously to be the precursors of regenerating olfactory receptor cells.

Animals↗

Addition of new cells to the olfactory bulb of adult zebrafish.

We have been examining patterns of cell proliferation in the brain of adult zebrafish. Understanding this process in fish may lead to important insights due to the tremendous regenerative capabilities of these animals. Fish were exposed to a 1% solution of the thymidine analog 5-bromo-2'-deoxyuridine (BrdU) for 1 hr before being returned to small aquaria that received frequent water changes. Animals were overanesthetized and perfused with Bouin's fixative solution after two survival periods (4 hr or 3-4 weeks). Paraffin immunohistochemistry using a monoclonal antibody against BrdU was used to visualize the newly generated cells. Quantitative analyses were performed on serial, 10-micron sections from 4 animals for each survival group. Statistical determinations were based on the nonparametric Mann-Whitney U test. The average number of BrdU-labeled nuclear profiles in the bulbs, from analysis of every 5th section, was significantly different between the two groups (22.0 +/- 6.8 [SEM] for the 4-hr group and 136.7 +/- 11.3 for the 4-wk group; p < 0.05). The volumes of the bulbs, however, were not different between the two groups (p > 0.5). These data indicate that either cells divided repeatedly during the longer survival period or cells migrated into the bulb from other brain regions. To examine this phenomenon further, the location of the new cells was analyzed in three mid-bulb sections (20 microns apart) from each animal. Both the area and number of labeled nuclei in each lamina were measured to obtain an average profile density. Comparison of the 4-hr and the 4-wk groups showed that density was significantly greater in all bulb layers in the long survival group (p < 0.05 for all). In the 4-hr survival group, cells were found mainly in the olfactory nerve layer. When examined after 4 wk, proportionately more labeled cells were found in the internal cell layer. This addition of new cells could be a result of neurogenesis, gliogenesis, and/or angiogenesis. We are currently performing double-labeling experiments to determine the types of cells that are added to the adult bulb. In addition, our future plans include investigating the origin of these cells and the signals that direct their formation.

Animals↗

[Nasal infection of the domestic cat with feline herpesvirus as an animal model of experimental viral labyrinthitis].

Looking for a suitable animal model of experimental labyrinthitis we performed in vivo infections of the domestic cat with feline rhinotracheitis virus. After instillation of the virus suspension into the nose of the anaesthetized cat we could find the complete virus particles in so-called virus factories in the nasal mucosa four days later by means of electron-microscopy. The adjacent ganglia of the olfactory nerve were stained by immunohistochemical methods and showed positive viral antigen. After embedding of the cochlea for electron microscopy virus particles could be demonstrated in the inflammatory cell material in the scala tympani. Our experimental model seems to be suitable for further studies of the rhinogenic way of infection in experimental viral labyrinthitis.

Animals↗

A review of 59 consecutive patients with lesions of the anterior cranial base operated on using the subcranial approach.

INTRODUCTION: Lesions of the anterior cranial fossa are still a challenge for the surgeon. Since Joram Raveh introduced the concept of a subcranial approach in 1978 it has been used in the treatment of lesions extending into the anterior cranial fossa. Our experience with the subcranial approach during the past 8 years at Turku University Central Hospital is described. PATIENTS: Fifty-nine consecutive patients underwent surgery using the subcranial approach for treating various benign and malignant neoplasms, for repair of frontobasal-midfacial fractures, and for repair of cerebrospinal fluid leaks. METHODS: Patient records were retrospectively reviewed, with special focus on surgical technique, early outcome, and complications. RESULTS: Nineteen of the patients were operated on because of combined fronto-naso-orbital and skull base fractures, 37 were tumour cases, and 3 patients required surgical repair for cerebrospinal fluid leakage. Significant complications consisted of two cases of meningitis. However, they were successfully treated with antibiotics. The most common late complaint was olfactory nerve dysfunction (44), other late complications such as diplopia (4), enophthalmos (2), scar tissue in the nasal cavity (2), and trigeminal nerve dysfunction (2) were also encountered. CONCLUSION: The subcranial approach affords exposure to the orbital, sphenoethmoidal, and clivus regions, as well as to the nasal and paranasal cavities. On the basis of this review, it is concluded that it is a safe and effective approach for treating lesions involving the anterior skull base.

Adolescent↗

Components of the plasma membrane of growing axons. III. Saxitoxin binding to sodium channels.

The density of sodium channels was measured in growing and mature axons of the olfactory nerve of the bullfrog, using as a probe the drug saxitoxin (STX). The toxin binds to control nerves from adult animals in a saturable manner with a dissociation constant of approximately 23 nM at 4 degrees C and a capacity of 72 fmol/mg wet weight, equivalent to about five sites per square micrometer of axolemma. In growing nerves, obtained from adult frogs 4-5 wk following removal of the original nerve, the STX-binding capacity per wet weight of tissue is markedly reduced, to approximately 25% of control values, and appears to decrease in the proximodistal direction. STX-binding data, expressed as STX/mg wet weight, was converted to STX/micron 2 of axolemma using stereologically derived values of membrane area per milligram wet weight of nerve. The axolemmal content (area/mg wet weight) of all regions of growing nerve is substantially decreased compared to controls, but increases in the proximodistal direction by 60%. These changes in axolemmal area result in calculated STX receptor densities (per unit axolemmal area) which, in distal regions, are approximately at the level of the mature nerve and, in proximal regions, are actually increased above controls by 50 to 70%. Upon comparing the axolemmal density of intramembrane particles, reported in the companion paper, with the calculated density of STX receptors in both mature and growing nerves, we find a correlation between STX receptors and intramembrane particles with diameters of 11.5-14.0 nm. The growing axon's gradient of sodium channels and the shift from this gradient to a uniform distribution in the mature axon suggest (a) that sodium channels are inserted into the perikaryal plasmalemma and diffuse from there into the growing axolemma, and (b) that the axolemma undergoes functional maturation during growth.

Amphibian Proteins↗

Membrane bistability in olfactory bulb mitral cells.

Whole-cell patch-clamp recordings were used to investigate the electrophysiological properties of mitral cells in rat main olfactory bulb brain slice preparations. The majority of mitral cells are bistable. These cells spontaneously alternate between two membrane potentials, separated by approximately 10 mV: a relatively depolarized potential (upstate), which is perithreshold for spike generation, and a relatively hyperpolarized potential (downstate), in which spikes do not occur. Bistability occurs spontaneously in the absence of ionotropic excitatory or inhibitory synaptic inputs. Bistability is voltage dependent; transition from the downstate to the upstate is a regenerative event activated by brief depolarization. A brief hyperpolarization can switch the membrane potential from the upstate to the downstate. In response to olfactory nerve (ON) stimulation, mitral cells in the upstate are more likely to fire an action potential than are those in the downstate. ON stimulation can switch the membrane potential from the downstate to the upstate, producing a prolonged and amplified depolarization in response to a brief synaptic input. We conclude that bistability is an intrinsic property of mitral cells that is a major determinant of their responses to ON input.

Action Potentials↗

The spatiotemporal analysis of odorants at the level of the olfactory receptor sheet.

Activity in two separate regions of the frog olfactory mucosa was sampled by simultaneously recording the summated neural discharges from the olfactory nerve branches originating from them. The difference in the activity from these two regions in response to a stimulus was measured by: (a) the ratio of the response amplitude recorded from the lateral nerve branch to that recorded from the medial nerve branch (LB/MB ratio), (b) the latency difference (or time interval) between these two responses. Equal concentrations of four different odorants were drawn into the nose by an artificially produced sniff of known dimensions. At each concentration in every animal the four chemicals were ranked in order of the magnitudes of their LB/MB ratios and again in order of their latency differences. Regardless of their concentration, the same chemicals fell into the same ranks in different animals. In addition, for each chemical the magnitudes of the ratios and latency differences showed only minimal changes with concentration. Thus, spatiotemporal patterns of relative response magnitudes and latency differences across the mucosa differentially represented the odorants. Such a spatiotemporal code, together with physicochemical considerations, suggested that the nose separates vapors in a manner similar to a gas chromatograph. This is further supported by the previously observed reversal of the ratio patterns with reversal of air flow direction through the olfactory sac.

Animals↗

Bulbar morphology and expression of bulbar dopamine and parvalbumin in experimentally-induced anosmic rats.

Morphological study was carried out in rats with olfactory dysfunction induced by deafferentation of serotonergic fibers in the olfactory bulb. With a computer capable of area measurements, olfactory bulbs of the anosmic rats were found to be decreased in size to 61% of control bulbs, and all bulbar layers were involved in the bulbar shrinkage. Given areas of each bulbar layer in control bulbs to be 100%, percentages of each bulbar layer in the anosmic rats were 23% in the olfactory nerve layer, 54% in the glomerular layer, 63% in the external plexiform layer, 83% in the internal plexiform layer and 81% in the granule cell layer. Dopamine-and parvalbumin-containing neurons were examined immunohistochemically in the experimentally-induced anosmic rats. As a result, immunoreactive neurons for these two chemical substances were significantly decreased in number (dopamine, 33% of control value; parvalbumin, 46% of control value). The present study, using an animal model of anosmia, provided quantitative data on the bulbar atrophy and showed effects of anosmia on expression of dopamine and parvalbumin in the bulb.

Animals↗

Neurite outgrowth promoting effects of enriched and mixed OEC/ONF cultures.

Olfactory ensheathing cell (OEC) transplants stimulate axon regeneration and partial functional recovery after spinal cord injury. However, it remains unclear whether enriched OEC or mixed transplants of OEC and olfactory nerve fibroblasts (ONF) are optimal for stimulating axon regrowth. The neurite outgrowth stimulating effects of enriched OEC, ONF, and mixed OEC/ONF cultures on neonatal cerebral cortical neurons were compared using co-cultures. We show that (1) OEC are more neurite outgrowth promoting than ONF, and (2) ONF do not enhance the neurite outgrowth stimulating effects of OEC in mixed OEC/ONF cultures. Hence, our data indicate that there is no preference for the use of enriched OEC or mixed OEC/ONF cultures with respect to stimulation of neurite growth in vitro.

Animals↗

Simple photoreceptors in Limulus polyphemus.

The "olfactory nerve," the endoparietal eye, and the rudimentary lateral eyes of Limulus (polyphemus) contain simple photoreceptor cells that duplicate many of the electrical responses of the retinular cells of the lateral eye; the responses are a receptor potential consisting of aninitial transient phase and a subsequent steady phase,low-amplitude fluctuations, and a small locally regenerative response to pulses of both light and current. Photic stimulation does not induce conducted action potentials, but does increase the membrane conductance. The receptor potentialrequires the presence of sodium ions in the external medium. Measurements of action and absorption spectra indicate a photopigment whose maximum absorption is of light with wavelength of 535 nanometers. The functional significance of these cells has not been ascertained.

Animals↗