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Uptake of inorganic mercury in the olfactory bulbs via olfactory pathways in rats.

Uptake and transport in the olfactory neurons may be an important means by which some heavy metals gain access to the brain. In the present study we explored whether inorganic mercury (203Hg2+) may be taken up in the CNS via the olfactory pathway. Autoradiography and gamma spectrometry showed that intranasal instillation of 203Hg2+ in the right nostrils of rats resulted in much higher levels of the metal in the right olfactory bulbs than in the left ones. At the side of the application of the 203Hg2+ there was also a labeling of the olfactory nerve bundles projecting to the olfactory bulbs as well as in the olfactory nerve-fibres constituting the olfactory nerve layer of the bulbs, which was not seen on the opposite side. The results also showed that the 203Hg2+ accumulated in the glomerular layer of the bulbs. These data indicate that our results can be ascribed to a movement of the mercury along the olfactory axons to their terminal parts in the glomeruli and not to circulatory uptake from the mucosal vasculature. At late survival intervals a low labeling was also discernable in the external plexiform layer, indicating that a low level of 203Hg2+ leaves the terminal arborizations of the axons in the glomeruli. An uptake of 203Hg2+ in the glomerular layer of the olfactory bulbs was also seen in rats given the metal intraperitoneally. This uptake was similar in the right and left bulbs and always much lower than in the right bulbs of the rats given 203Hg2+ in the right nostrils. The intraperitoneal injections in addition resulted in an uptake of the 203Hg2+ in the olfactory epithelium. We propose that in these rats the mercury is taken up from the blood into the olfactory neurons and then moves along the axons to their terminations in the olfactory bulbs. In humans a continuous exposure of the nasal cavity to mercury vapor (Hg0), released from amalgam fillings and oxidized to Hg2+ in the olfactory mucosa, as well as a potential uptake of Hg2+ in the olfactory neurons from the blood, may lead to considerable concentrations of the metal in the olfactory bulbs.

Animals↗

Cellular and subcellular localization of gamma-aminobutyric acidB receptors in the rat olfactory bulb.

Olfactory nerve axons terminate in rounded regions of the olfactory bulb, termed glomeruli, where they make excitatory synapses with the dendrites of second-order neurons. Neurotransmission from the olfactory nerve to the postsynaptic targets is negatively regulated by gamma-aminobutyric acid (GABA), and there is evidence that inhibition of sensory input is mediated, at least in part, by GABA(B) receptors. Using an antiserum that recognizes two GABA(B) receptor splice variants (GBR1a and GBR1b), we show here that GABA(B) receptors are located on the axon terminals of the olfactory nerve, where they are concentrated at sites of axodendritic apposition. Taken with previous data, these results indicate that GABA(B) receptors act presynaptically to regulate the release of glutamate from olfactory nerve terminals.

Alternative Splicing↗

Infusion of the metabotropic receptor agonist, DCG-IV, into the main olfactory bulb induces olfactory preference learning in rat pups.

DCG-IV, a type 2 metabotropic glutamate receptor (mGluR2) agonist, was infused into the main olfactory bulb of 1-week-old pups exposed to peppermint odor. A preference for peppermint was demonstrated 24 h later. The data support the proposal that disinhibition at dendrodendritic synapses between granule cells and mitral cells is a critical component of olfactory memory formation in the rat pup olfactory bulb as well as in the accessory olfactory bulb of adult rodents.

Animals↗

NMDA-receptor modulation of lateral inhibition and c-fos expression in olfactory bulb.

Olfactory bulb primary output neurons, mitral/tufted cells, are glutamatergic and excite inhibitory interneurons, granule cells, by activation of both alpha-amino-3-hydroxy-5-methyl-ioxazole-4-propionic acid (AMPA) and N-methyl-D-aspartate (NMDA) glutamate receptors. The data presented here demonstrate that the NMDA antagonists MK-801 and CGP39551, but not ketamine, significantly enhanced expression of c-fos mRNA by mitral cells as measured by in situ hybridization. All three antagonists significantly reduced mitral/tufted cell excitation of granule cells as measured with extracellular field potentials following antidromic stimulation of the lateral olfactory tract (LOT). In turn, the NMDA antagonists significantly reduced granule cell mediated feedback inhibition of mitral/tufted cells, as measured with field potential recordings of paired-pulse LOT stimulation, suppression of mitral/tufted cell single-unit spontaneous activity following LOT stimulation, and intracellularly recorded IPSP amplitude in mitral/tufted cells following LOT stimulation. While there was not a perfect correlation between the effects of the NMDA antagonists on c-fos mRNA expression and on inhibition, the results suggest that disinhibition of mitral/tufted cells accounts for the observed enhancement in c-fos mRNA expression induced by NMDA receptor antagonists.

2-Amino-5-phosphonovalerate↗

Aging in the rat olfactory system: relative stability of piriform cortex contrasts with changes in olfactory bulb and olfactory epithelium.

Previous studies have quantified growth and atrophy of the olfactory bulb and olfactory epithelium of the Sprague-Dawley rat from maturity to senescence. Major events occurring in these structures include changes in the volume of mitral cells and changes in the number of septal olfactory receptors. These effects are large, consist of a growth phase followed by atrophy, and are temporally related in that events in the olfactory epithelium precede those in the mitral cells. A hypothesis of aging based on transneuronal effects would predict that these changes would be similarly transmitted to the next synaptic station in the olfactory pathway. Therefore, cells and synapses of the piriform cortex were studied in rats 3, 12, 18, 24, 27, 30, and 33 months of age. Alternate Vibratome sections through brains perfused with mixed aldehydes were processed for light and electron microscopy. No significant age effects were found for the volumes of cortical laminae Ia and Ib. Both numerical and surface density of synaptic apposition zones in layer Ia, formed primarily by mitral cell axons, were stable with age. A modest (18%) but significant decline in the proportion of layer Ia occupied by dendrites and spines was mirrored by an increase in the proportion of glial processes; no change in the proportion of axons and terminals was observed. Neither nuclear volume, nor soma volume, nor numerical density of layer II neurons changed with age. Thus, contacts made in the piriform cortex by mitral cell axons remain relatively stable in senescence, despite the marked volumetric changes in the mitral cell somata, changes which were confirmed again in this study. Age-related dendritic regression in layer II neurons may be attributable to functional deafferentation subsequent to reduced receptor input to mitral cells.

Age Factors↗

Transgenic expression of Cre recombinase in mitral/tufted cells of the olfactory bulb.

Olfactory information is conveyed from the periphery to the olfactory cortices through mitral and tufted (M/T) cells in the olfactory bulb. A mouse with a specific expression of Cre recombinase in M/T cells is essential for genetic marking of M/T cells and manipulating their properties. Protocadherin 21 (Pcdh21) expression is highly restricted to M/T cells. Here we report a transgenic mouse line, Pcdh21-Cre, in which 10-kb mouse Pcdh21 promoter drives the expression of Cre recombinase. In Pcdh21-Cre mice, Cre recombinase activity is predominantly detected in M/T cells, visualized with the anti-CFP immunostaining in offspring of a cross between Pcdh21-Cre and the reporter Rosa26-loxP-stop-loxP-CFP strain. These results demonstrate that the 10-kb Pcdh21 promoter can drive transcription in M/T cells and Pcdh21-Cre mice can be used to excise floxed DNA fragments in M/T cells, which provides a valuable tool to reveal the structure and function of the central olfactory circuits.

Animals↗

Axon-glia communication evokes calcium signaling in olfactory ensheathing cells of the developing olfactory bulb.

Olfactory ensheathing cells (OECs) accompany receptor axons in the olfactory nerve and promote axonal growth into the central nervous system. The mechanisms underlying the communication between axons and OECs, however, have not been studied in detail yet. We investigated the effect of activity-dependent neuronal transmitter release on Ca(2+) signaling of OECs in acute mouse olfactory bulb slices using confocal Ca(2+) imaging. TTX-sensitive axonal activity upon electrical nerve stimulation triggers a rise in cytosolic Ca(2+) in OECs, which can be mimicked by application of DHPG, an agonist of metabotropic glutamate receptors (mGluRs). Both stimulation- and DHPG-induced Ca(2+) transients in OECs were abolished by depletion of intracellular Ca(2+) stores with cyclopiazonic acid (CPA). The mGluR(1)-specific antagonist CPCCOEt completely inhibited DHPG-evoked Ca(2+) transients, but reduced stimulation-induced Ca(2+) transients only partly, suggesting the involvement of another neurotransmitter. Application of ATP evoked CPA-sensitive Ca(2+) transients in OECs, which were inhibited by the P2Y(1)-specific antagonist MRS2179. Co-application of CPCCOEt and MRS2179 almost completely blocked the stimulation-induced Ca(2+) transients, indicating that they were mediated by mGluR(1) and P2Y(1) receptors. Our results show that OECs are able to respond to olfactory nerve activity with an increase in cytosolic Ca(2+) due to glutamate and ATP release.

Adenosine Triphosphate↗

Viral tracing identifies distributed columnar organization in the olfactory bulb.

Olfactory sensory neurons converge onto glomeruli in the olfactory bulb (OB) to form modular information processing units. Similar input modules are organized in translaminar columns for other sensory modalities. It has been less clear in the OB whether the initial modular organization relates to a columnar structure in the deeper layers involved in local circuit processing. To probe synaptic connectivity in the OB, we injected a retrograde-specific strain of the pseudorabies virus into the rat OB and piriform cortex. The viral-staining patterns revealed a striking columnar organization that extended across all layers of the OB from the glomeruli to the deep granule cell layer. We hypothesize that the columns represent an extension of the glomerular unit. Specific patterning was observed, suggesting selective, rather than distance-dependent, center-surround connectivity. The results provide a previously undescribed basis for interpreting the synaptic connections between mitral and granule cells within the context of a columnar organization in the OB and have implications for olfactory coding and network organization.

Animals↗

[Regulation of c-Fos expression in the rat olfactory bulbs during olfactory learning].

The distribution of c-Fos-immunopositive neurons was examined in the mitral/tufted and granular cell layers in the medium part of the main olfactory bulbs of 18-day-old rats after they had been trained for propionic acid vapour-guided search for dam in the Y-maze. On the next day these pups exhibited a strong preference for the propionic acid odor as compared to the control pups trained for this task without the odor cue and odor-familiarized pups exposed to propionic acid as a novel neutral stimulus. Exposure to propionic acid produced a moderate activation of c-Fos expression, mainly in the granular layer of the dorsomedial part of the bulb. Training in the Y-maze devoid of odor cues resulted in diffuse increase in the number of c-Fos-positive neurons both in the mitral and granular cell layers in all parts of the olfactory bulb. Maze training with the odor cue produced activation of c-Fos expression (which significantly exceeded the non-odor Y-maze group) in the dorsomedial olfactory bulb. These data suggest that associative olfactory conditioning results in activation of c-Fos expression that combines the effect of diffuse motivational excitation and specific olfactory input to the neurons which process odor cues.

Animals↗

The differential projections of the olfactory bulb and accessory olfactory bulb in mammals.

Three species were studied, the rabbit, opossum and rat. Lesions of the main olfactory bulb caused terminal degeneration, assayed by the Fink-Heimer method, to occur in the ipsilateral olfactory tubercle, prepyriform cortex (including its periamygdaloid part), ventrolateral entorhinal area, and in anterior and posterolateral divisions of the cortical amygdaloid nucleus. The various parts of the ipsilateral anterior olfactory nucleus and the rostroventral end of the anterior continuation of the hippocampus (hippocampal rudiment) also received this projection. Lesions of the accessory olfactory bulb, which receives its sensory input from the vomeronasal (Jacobson's) organ, caused terminal degeneration to occur in the medial amygdaloid nucleus and in a posteromedial part of the cortical amygdaloid nucleus. This projection was conveyed by an accessory olfactory tract, which is accompanied in part of its course by a small nucleus, the bed nucleus of the accessory olfactory tract. The accessory olfactory tract is initially a part of the lateral olfactory tract but becomes increasingly indivuated at more posterior levels. It parts company with the lateral olfactory tract at the rostral end of the amygdaloid region, and, in addition to distributing to the medio-cortical amygdaloid region, it enters the stria terminalis to terminate in the bed nucleus of the stria terminalis in a small region bearing cytoarchitectonic resemblance to the medial amygdaloid nucleus. The topographic segregation of the areas of termination of the olfactory and accessory olfactory (vomeronasal) projections is suggestive of a functional dichotomy in the organization of the olfactory system...

Amygdala↗

Retrograde transport of nerve growth factor from olfactory bulb to olfactory epithelium.

NGF is produced in the olfactory bulb and transported from the bulb to cholinergic neurons in the horizontal limb of the diagonal band. Although the expression of NGF receptors has been reported in olfactory epithelial cells, no correlation between NGF and olfactory epithelial cells has been found. This study aimed to define whether or not retrograde transport of NGF occurs in the olfactory neural system using the method of radioluminography. 125I-labeled NGF injected into the olfactory bulb was taken up and transported to the olfactory epithelium 18 h after injection. This finding suggests that bulbar NGF may act as a neurotrophic factor in olfactory epithelial cells.

Animals↗

The olfactory bulb and olfactory mucosa obtained from human cadaver donors as a source of olfactory ensheathing cells.

During the last decade, olfactory ensheathing cells (OECs) have been successfully applied in multiple experimental approaches aimed to repair damaged mammalian spinal cord. Some of these experiments have consequently been translated into clinical trials. Finding a reliable source of human OECs that is easily accessible and can ensure a sufficient number of cells is a major prerequisite for conducting studies on OEC-mediated spinal cord regeneration. Here, we present a procedure for obtaining olfactory bulbs (OBs) and olfactory mucosa (OM) simultaneously from adult cadaver heart-beating donors for OEC isolation and analyze some of the factors that may condition successful OEC culture. We show that the results of OEC culture from OBs (10 cases) correlated significantly with warm ischemia time (WIT) as well as the initial viability of the isolated cells. Efficient OEC culture was possible when the WIT for the OB was up to 20 min. Brain damage, assessed by determination of S100B serum level, was not related to the success of OEC culture from the OB. Cadaver OM (7 cases) was shown to be a more reliable source of human OECs than the OB. In most of the examined cases the efficacy of culturing OECs from cadaver OM obtained even 180 min after cardiac arrest was comparable to that of living patients. The method of obtaining OBs and OM from cadavers enables the use of an alternative source of primary adult human OECs for further preclinical and clinical studies on their neurotrophic properties.

Adult↗

Differential projections from locus coeruleus to olfactory bulb and olfactory tubercle: an HRP study.

The microiontophoretic administration of horseradish peroxidase (HRP) to the olfactory bulb (OB) or olfactory tubercle (OT) in cats and rats yielded similar results in both species. After an OB HRP-injection ipsilateral and contralateral labelled neurons were seen in the piriform cortex, polymorphic layer of OT, magnocellular preoptic region, lateral hypothalamus, ventromedial hypothalamic nucleus and locus coeruleus (LC). In both species more labelled structures were found after an OT HRP-injection than after an OB HRP-injection. The substantia nigra in rats was more abundantly labelled after an OT injection than after an OB one. In cats the dorsal and the ventral raphe were also labelled. In either species, OT HRP-injections resulted in a higher frequency of LC labelled neurons than after OB injections. These results favor the hypothesis that the OT plays an important role as a relay station for efferent inflow from the brain stem en route to the OB.

Animals↗

Dendritic cell-like immunoreactivity in the glomerulus of the olfactory bulb and olfactory nerves of mice.

Dendritic cell-like immunoreactivity was examined in the mouse brain. The glomerulus of the olfactory bulb and the olfactory nerves were stained by antibodies against the dendritic cells, NLDC-145 and MIDC-8, while these structures were not stained by antibodies against microglia or macrophages, F4/80, Mac1 or CD45. Immunoelectron microscopy showed that the immunoreaction for NLDC-145 was localized to the sheath and presynaptic terminals of the olfactory nerves. These findings suggest that the sheath and presynaptic terminals of the primary olfactory nerves have some degree of the antigenicity in common with dendritic cells.

Animals↗

Inputs from the olfactory bulb and olfactory cortex to the entorhinal cortex in the cat. I. Anatomical observations.

The spatial organization and laminar distribution of projections from the olfactory bulb and the anterior (PPCa) and posterior (PPCp) divisions of the prepiriform cortex to the entorhinal cortex were studied with anterograde (3H-leucine) and retrograde (WGA-HRP) tracing techniques. After 3H-leucine injections into the olfactory bulb transported labeling was seen over the lateral entorhinal area, except its most medial part, and over the rostral part of the medial entorhinal area. The labeling covers exclusively layer Ia. The lateral and medial entorhinal areas are also reached by fibers from the prepiriform cortex. The projection to the medial entorhinal area has not been described previously. Following injections of 3H-leucine into the PPCa transported labeling is present over the entire expanse of the entorhinal cortex and is located over layer Ib with the greatest density in its superficial part. Injections of 3H-leucine into the PPCp give rise to transported labeling over much of the entorhinal cortex. No labeling was found over the most medial parts of the medial subdivision (VMEA) of the lateral entorhinal area and the medial entorhinal area. Labeling occupies layer Ib, especially its middle part, and layers II and III. Both PPCa and PPCp appear to project most heavily to the dorsal (DLEA) and ventral (VLEA) subdivisions of the lateral entorhinal area. From the retrograde experiments it can be inferred that cells of layers II and III of the PPCa project predominantly to the DLEA, whereas those of the PPCp project predominantly to the VLEA. The MEA receives its heaviest projection from layer II of both PPCa and PPCp.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Olfactory marker protein modulates primary olfactory axon overshooting in the olfactory bulb.

Olfactory marker protein (OMP) is expressed by mature primary olfactory sensory neurons during development and in adult mice. In mice that lack OMP, olfactory sensory neurons have perturbed electrophysiological activity, and the mice exhibit altered responses and behavior to odor stimulation. To date, defects in axon guidance in mice that lack OMP have not been investigated. During development of the olfactory system in mouse, primary olfactory axons often overshoot their target glomerular layer and project into the deeper external plexiform layer. These aberrant axonal projections are normally detected within the external plexiform layer up to postnatal day 12. We have examined the projections of primary olfactory axons in OMP-tau:LacZ mice and OMP-GFP mice, two independent lines in which the OMP coding region has been replaced by reporter molecules. We found that axons overshoot their target layer and grow into the external plexiform layer in these OMP null mice as they do in wild-type animals. However, in the absence of OMP, overshooting axons are more persistent and remain prominent until 5 weeks postnatally, after which their numbers decrease. Overshooting axons are still present in these mice even at 8 months of age. In heterozygous mice, axons also overshoot into the external plexiform layer; however, there are fewer axons, and they project for shorter distances, compared with those in a homozygous environment. Our results suggest that perturbed electrophysiological responses, caused by loss of OMP in primary olfactory neurons, reduce the ability of primary olfactory axons to recognize their glomerular target.

Animals↗

Distribution and morphology of substance P-immunoreactive structures in the olfactory bulb and olfactory peduncle of the common marmoset (Callithrix jacchus), a primate species.

The present study describes the morphology and distribution of substance P-immunoreactive (SP-ir) elements in the olfactory bulb (OB) and olfactory peduncle (OP) of the common marmoset (Callithrix jacchus), a primate species. SP-ir neurons are very abundant in the OB and belong to two types. External tufted cells are present in the glomerular layer (GL), whereas granule cells are found in the deeper layers, especially in the granule cell layer (GRL), but also scattered in the OP. SP-ir fibers, putatively of central origin, were identified in the OP. They ascend into the bulbar layers. The SP-chemoarchitecture of the marmoset OB and OP does not differ more from rat, guinea pig and cat, than the SP-chemoarchitecture of these species varies among one another.

Animals↗

Comparison of cell populations derived from canine olfactory bulb and olfactory mucosal cultures.

OBJECTIVE: To evaluate the numbers and proportions of olfactory ensheathing cells (OECs) in cell cultures derived from the olfactory bulb (OB) and olfactory mucosa of dogs. ANIMALS: 7 dogs. PROCEDURES: OB tissue and olfactory mucosa from the nasal cavity and frontal sinus were obtained from euthanatized dogs and prepared for cell culture. At 7, 14, and 21 days of culture in vitro, numbers and proportions of OECs, astrocytes, and fibroblasts were determined via immunocytochemistry. Antibody against the low-affinity nerve growth factor receptor p75 was used to identify OECs, antibody against glial fibrillary acidic protein was used to identify astrocytes, and antibody against fibronectin was used to identify fibroblasts. RESULTS: Cultured OECs derived from the olfactory mucosa of the nasal cavity and frontal sinus had similar characteristics. However, whereas OECs in the OB cell cultures constituted approximately 50% of the cells at 7 days and approximately 75% at 21 days the proportion of OECs in cultures derived from both mucosal types was much lower, with approximately 40% OECs at 7 days and approximately 25% at 21 days. Analysis of OEC numbers revealed that these changes were accompanied by corresponding decreases and increases in the population of cells with fibronectin receptors. CONCLUSIONS AND CLINICAL RELEVANCE: Although olfactory mucosal cell cultures yielded a sufficient number of OECs for spinal cord transplantation procedures in dogs, modification of culture conditions would be required to ensure that the derived cell population contained a sufficient proportion of OECs.

Animals↗