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PubMed2003

Detection of pathologic prion protein in the olfactory epithelium in sporadic Creutzfeldt-Jakob disease.

BACKGROUND: Olfactory cortexes and the olfactory tracts are involved in sporadic Creutzfeldt-Jakob disease. We examined peripheral regions of the olfactory sensory pathway, including the olfactory mucosa, to assess whether pathologic infectious prion protein (PrPSc) is deposited in the epithelium lining the nasal cavity. METHODS: We studied nine patients with neuropathologically confirmed sporadic Creutzfeldt-Jakob disease. We obtained the brain, the cribriform plate with the attached olfactory mucosa, and the surrounding respiratory epithelium at autopsy. Control samples of nasal mucosa were obtained post mortem or at biopsy from age-matched control subjects and from control patients with other neurodegenerative diseases. The olfactory and respiratory mucosa and the intracranial olfactory system were analyzed by light microscopy, immunohistochemistry, and Western blotting for pathological changes and for deposition of PrPSc. RESULTS: In all nine patients with sporadic Creutzfeldt-Jakob disease, PrPSc was found in the olfactory cilia and central olfactory pathway but not in the respiratory mucosa. No PrPSc was detected in any of the tissue samples from the 11 controls. CONCLUSIONS: Our pathological and biochemical studies show that PrPSc is deposited in the neuroepithelium of the olfactory mucosa in patients with sporadic Creutzfeldt-Jakob disease, indicating that olfactory biopsy may provide diagnostic information in living patients. The olfactory pathway may represent a route of infection and a means of spreading prions.

Antibodies, Monoclonal↗
PubMed1993

Monoclonal antibody labels olfactory and visual pathways in Drosophila and Apis brains.

We employed a monoclonal antibody raised against Drosophila brain homogenate for a comparative immunocytochemical analysis of visual and olfactory pathways in brains of two insect species. On Western blots of Drosophila and Apis nervous tissue, antibody fb45 recognized an antigen with an apparent molecular weight higher than 180 kD. Application of the antibody to sections of Drosophila and Apis brain stained certain interneurons which conspicuously fasciculate in common tracts or neuropilar compartments. Both in Drosophila and in Apis, the antigen was also expressed on the perineural sheath and granular cell compartments in the majority of neuronal cell bodies. The antibody stained monopolar cells in the visual system of both species, and in Apis those fibers of the anterior superior optic tract which link the medulla with the mushroom bodies. In Drosophila, bundles of Kenyon cells of the mushroom bodies were stained. In worker bees and drones, the relay neurons of the median and lateral antennoglomerular tracts were labelled. Since the recognition of the antigen does not require fixation, the antibody can be employed to label selectively living neurons in dissociated cell culture. This opens up the possibility for future functional studies on the role of the antigen in vitro.

Animals↗
PubMed2005

Delayed olfactory nerve regeneration in ApoE-deficient mice.

Apolipoprotein E (apoE), a lipid transporting protein, is extensively expressed in the primary olfactory pathway, but its function is unknown. We previously reported increased apoE levels in the olfactory bulb (OB) following olfactory epithelium (OE) lesion in mice, and hypothesized that apoE may play a vital role in olfactory nerve (ON) regeneration. To directly test this hypothesis, we examined the rate of ON regeneration following OE lesion in apoE deficient/knockout (KO) and wild-type (WT) mice. OE was lesioned in 2- to 3-month-old mice by intranasal irrigation with Triton X-100 (TX). OB were collected at 0, 3, 7, 21, 42, and 56 days post-lesion. OB recovery was measured by both immunoblotting and immunohistochemical analysis of growth cone associated protein (GAP) 43 and olfactory marker protein (OMP). The results revealed that (1) OMP recovery in the OB was significantly slower in apoE KO compared to WT mice; (2) recovery of glomerular area was similarly slower; and (3) GAP43 increases and return to prelesion levels in the OB were slower in KO mice. Together, these results show that olfactory nerve regeneration is significantly slower in KO mice as compared to WT mice, suggesting apoE facilitates olfactory nerve regeneration.

Animals↗
PubMedDate not supplied

Heterogeneity in olfactory neurons in mouse revealed by differential expression of glycoconjugates.

Cell surface glycoconjugates have been implicated in the growth and guidance of subpopulations of primary olfactory axons. While subpopulations of primary olfactory neurons have been identified by differential expression of carbohydrates in the rat there are few reports of similar subpopulations in the mouse. We have examined the spatiotemporal expression pattern of glycoconjugates recognized by the lectin from Wisteria floribunda (WFA) in the mouse olfactory system. In the developing olfactory neuroepithelium lining the nasal cavity, WFA stained a subpopulation of primary olfactory neurons and the fascicles of axons projecting to the target tissue, the olfactory bulb. Within the developing olfactory bulb, WFA stained the synaptic neuropil of the glomerular and external plexiform layers. In adults, strong expression of WFA ligands was observed in second-order olfactory neurons as well as in neurons in several higher order olfactory processing centres in the brain. Similar, although distinct, staining of neurons in the olfactory pathway was detected with Dolichos biflorus agglutinin. These results demonstrate that unique subpopulations of olfactory neurons are chemically coded by the expression of glycoconjugates. The conserved expression of these carbohydrates across species suggests they play an important role in the functional organization of this region of the nervous system.

Animals↗
PubMed1991

Pathologic changes in olfactory neurons in Alzheimer's disease.

Olfactory deficits and degenerative changes in central olfactory pathways are prominent in patients with Alzheimer's disease (AD). We hypothesized that peripheral olfactory neurons that reside in the nasal epithelium would show degenerative changes similar to the characteristic pathologic features of AD brain. Immunohistochemical studies of nasal tissue taken at autopsy reveal extensive degeneration in the sensory epithelium as well as abnormal neurites that share immunoreactive epitopes with dystrophic neurites and neurofibrillary tangles of the AD brain. The neuritic masses are stained with well-characterized monoclonal antibodies that do not normally stain olfactory neurons but which are very reactive with dystrophic neuritic structures and neurofibrillary tangles in AD brain. These include antibodies to phosphorylated and nonphosphorylated neurofilament subunits, tau, and also ALZ50, which is characteristically reactive with AD but not with normal brains. Such changes are present in 81% of AD patients. Similar accumulations of ectopic neurites are found in the olfactory epithelium of about 22% of non-demented patients. Preliminary statistical analysis fails to reveal any age-linked association. It has been proposed that the aged monkey is a good model for AD inasmuch as amyloid accumulations similar to those of humans are found in monkey brain. We examined a series of 13 rhesus monkeys, including aged animals with behavioral deficits. Although the olfactory epithelium was very similar to that of humans, no abnormal olfactory structures were observed. Aged rhesus monkeys do not appear to be a good model for the neuritic abnormalities of AD.

Alzheimer Disease↗
PubMed2004

Statistical mapping of functional olfactory connections of the rat brain in vivo.

The olfactory pathway is a unique route into the brain. To better characterize this system in vivo, rat olfactory functional connections were mapped using magnetic resonance (MR) imaging and manganese ion (Mn2+) as a transport-mediated tracer combined with newly developed statistical brain image analysis. Six rats underwent imaging on a 1.5-T MR scanner at pre-administration, and 6, 12, 24, 36, 48, and 72 h and 5.5, 7.5, 10.5, and 13.5 days post-administration of manganese chloride (MnCl2) into the right nasal cavity. Images were coregistered, pixel-intensity normalized, and stereotactically transformed to the Paxinos and Watson rat brain atlas, then averaged across subjects using automated image analysis software (NEUROSTAT). Images at each time point were compared to pre-administration using a one-sample t statistic on a pixel-by-pixel basis in 3-D and converted to Z statistic maps. Statistical mapping and group averaging improved signal to noise ratios and signal detection sensitivity. Significant transport of Mn2+ was observed in olfactory structures ipsilateral to site of Mn2+ administration including the bulb, lateral olfactory tract (lo) by 12 h and in the tubercle, piriform cortex, ventral pallidum, amygdala, and in smaller structures such as the anterior commissure after 24 h post-administration. MR imaging with group-wise statistical analysis clearly demonstrated bilateral transsynaptic Mn2+ transport to secondary and tertiary neurons of the olfactory system. The method permits in vivo investigations of functional neuronal connections within the brain.

Amygdala↗
PubMed1982

Mitral cell degeneration and sensory function in the neurological mutant mouse Purkinje cell degeneration (PCD).

Selective degeneration of mitral cells, the principal relay neuron in the central olfactory pathway, is described in the mutant mouse Purkinje cell degeneration (PCD). Concomitant effects are described in adjacent layers of the olfactory bulb which suggest a reorganization of local neuronal circuits. The 2-deoxyglucose technique produced results which suggested that the topographical distribution of sensory afferent input to the bulb was not affected.

Animals↗
PubMedDate not supplied

Olfactory thresholds are associated with degree of dementia in Alzheimer's disease.

Recent neuroanatomical studies have noted that regions of the olfactory pathways contain high levels of neuritic plaques and neurofibrillary tangles, pathological hallmarks of Alzheimer's disease; and that the olfactory epithelium, the most peripheral level of the system, exhibits anatomical and biochemical changes in Alzheimer's disease. The present experiments investigated thresholds for olfactory and taste stimuli in patients with Probable Alzheimer's disease. Olfactory thresholds of Alzheimer's patients were significantly elevated relative to controls and were significantly correlated with scores on dementia scales. Taste thresholds of Alzheimer's patients were within normal limits and unrelated to scores on dementia scales. These results suggest that increased olfactory thresholds in patients with Alzheimer's disease reflect the effects of the disease process and, thus, may aid in the diagnosis and in the understanding of Alzheimer's disease.

Aged↗
PubMed1987

Olfactory marker protein is present in olfactory receptor cells of human fetuses.

Light microscopic observations indicated that cellular differentiation of the primary olfactory pathway in human fetuses was relatively advanced by the end of the first trimester. However, immunohistochemical staining showed that olfactory marker protein was present in the receptor cells only at about 28 weeks post-conception. At about 32 weeks, modest amounts of the protein could be found in the peripheral olfactory nerve layer and a few glomeruli, with increased staining observed in subsequent weeks. The results show conclusively that human fetal olfactory neurons synthesize olfactory marker protein during the later half of gestation.

Central Nervous System↗
PubMed1999

Internalization of G protein-coupled receptors in single olfactory receptor neurons.

Desensitization of many G protein-coupled receptors after ligand binding generally involves phosphorylation of the receptors and internalization of the ligand-bound, phosphorylated receptors by a clathrin-mediated endocytic pathway. Olfactory receptor neurons from the channel catfish (Ictalurus punctatus) express the G protein-coupled odorant receptors and metabotropic glutamate receptors. To determine whether a clathrin-dependent receptor internalization pathway exists in olfactory receptor neurons, western blotting and immunocytochemistry were used to identify and localize clathrin and dynamin in isolated olfactory neurons. Clathrin and dynamin immunoreactivity was found in the cell bodies, dendrites, and dendritic knobs of the neurons. Using the activity-dependent fluorescent dye FM1-43 to monitor receptor internalization, we show that single olfactory neurons stimulated with the odorant amino acid L-glutamate internalized the dye. Odorant-stimulated neurons showed a consistent pattern of internalized FM1-43 fluorescence localized in the cell bodies and dendritic knobs. Odorant-stimulated internalization was unaffected by the caveolae activator okadaic acid and was significantly decreased by a metabotropic glutamate receptor antagonist, suggesting that a functional, clathrin-dependent, receptor-mediated internalization pathway exists in olfactory receptor neurons.

Animals↗
PubMed2005

5-Hydroxytryptamine action in the rat olfactory bulb: in vitro electrophysiological patch-clamp recordings of juxtaglomerular and mitral cells.

The olfactory bulb, first relay of olfactory pathways, is densely innervated by serotoninergic centrifugal fibers originating from the raphe nuclei. Although serotonin innervation was reported to be involved in olfactory learning in mammals, the action of this neurotransmitter on its putative cellular targets has been never described through unitary recordings. This lack of data initiated the present study where the effects of 5HT on juxtaglomerular and mitral cells are analyzed using whole-cell recordings on olfactory bulb slices. Serotonin depolarizes 34% of 525 JG cells. A multivariate statistical analysis of juxtaglomerular cells characteristics shows that the serotonin responsive cell group can be individualized regarding their tonic discharge-mode in response to a direct current injection, their lower expression of hyperpolarization-activated cation current and their low membrane capacities. The use of ion channel blockers and ramp voltage protocol indicate that serotoninergic depolarization of juxtaglomerular cells may be due to a nonselective cation current with a reversal potential of -44 mV. Pharmacological tests with serotonin receptor antagonists and agonists reveal that 5HT action on juxtaglomerular cells would be mainly mediated by 5HT2C receptors. In mitral cells, serotonin acts on 49.1% of the 242 tested cells, inducing two types of responses. A first subset of mitral cells (26.8%, n=65) were hyperpolarized by serotonin. This response would be indirect and mediated by action of GABA on GABAA receptors since it was antagonized by bicuculline. The involved GABAergic neurons are hypothesized to be juxtaglomerular and granular cells, on which serotonin would act mainly via 5HT2C and via 5HT2A receptors respectively. The second subset of mitral cells (22.3%, n=54) were directly depolarized by serotonin acting through 5HT2A receptors. Our data on serotonin action on juxtaglomerular cells and mitral cells reveal a part of functional mechanisms whereby serotonin can act on olfactory bulb network. This is expected to enrich the understanding of its determining role in olfactory learning.

4-Aminopyridine↗
PubMed1979

Failure of deodorized males to induce oestrus in the wild mouse.

The ability of female mice to return to oestrus following exposure to males perfumed either with oil of wintergreen or with the commercial perfume, "Kanta" was evaluated. Unisexual grouping of female mice induced anoestrus in all individuals. Oestrus was, however, promptly induced in the majority of unisexually grouped females by exposure to normal males. By contrast, exposure to perfumed males failed to induce oestrus in unisexually grouped females. The results suggest that male urine which is the source of the primer pheromone involved in the induction of oestrus was ineffective because of the masking effect of artificial scents. Hence the unisexually grouped females were unable to perceive the pheromone from males and continued to remain in anoestrus following exposure to perfumed males. The results provide additional evidence in support of the view that the urinary pheromone produced by males induces oestrus in females by acting through olfactory pathways.

Animals↗
PubMed1999

Regional distribution and cell type-specific expression of the mouse F3 axonal glycoprotein: a developmental study.

The expression of the mouse axonal adhesive glycoprotein F3 and of its mRNA was studied on sections of mouse cerebellar cortex, cerebral cortex, hippocampus, and olfactory bulb from postnatal days 0 (P0) to 30 (P30). In cerebellar cortex, a differential expression of F3 in granule versus Purkinje neurons was observed. F3 was highly expressed during migration of and initial axonal growth from cerebellar granule cells. The molecule was then downregulated on cell bodies and remained expressed, although at low levels, on their axonal extensions. On Purkinje cells, F3 was strongly expressed on cell bodies and processes at the beginning of the second postnatal week; by P16 it was restricted to neurites of Purkinje cells subpopulations. In the cerebral cortex, the molecule was highly expressed on migrating neurons at P0; by P16, it was found essentially within the neuropil with a diffuse pattern. In the hippocampal formation, where F3 was expressed on both pyramidal and granule neurons, a clear shift from the cell bodies to neurite extensions was observed on P3. In the olfactory pathway, F3 was expressed mainly on olfactory nerve fibers, mitral cells, and the synaptic glomeruli from P0 to P3, with a sharp decline from P11 to P16. As a whole, the data show that F3 protein expression is regulated at the regional, cellular, and subcellular levels and suggest that, in different regions, it can be proposed as a reliable neuronal differentiation marker.

Aging↗
PubMed1997

Rapid classical conditioning of odor response in a physiological model for olfactory research, the tiger salamander.

In recent years there have been a number of important advances in the understanding of cellular mechanisms related to olfactory function. Hypotheses regarding the complex relationships among odorant structure, physiological activity and behavioral outcome generated by these findings, however, remain largely untested due to a paucity of psychophysical data on stimulus discrimination in the same experimental species. Comparisons between behavioral and physiological responses are essential for elucidating the critical aspects of stimulus coding in sensory systems. We have developed a method for generating psychophysical data in one of the primary model species used in olfactory research, the tiger salamander, Ambystoma tigrinum. These psychophysical experiments are carried out under the same conditions as physiological experiments in our laboratory. Using classical conditioning, individual salamanders are trained over a period of 2-3 h to show skin potential responses to odor and not air. Failure to train using backward pairing demonstrates that the response is not due to sensitization or pseudoconditioning. The conditioned response is mediated by the olfactory pathway, as it is blocked by olfactory nerve section. We show that salamanders detect three odorants that are commonly used stimuli in physiological experiments (butyl alcohol, butyl acetate and amyl acetate), but cannot detect a fourth common experimental odorant, camphor. This method should be a powerful tool for studying olfactory information processing by providing data on discriminability of stimuli used in salamander physiological studies.

Acetates↗
PubMed1987

Trigeminal and olfactory synergism in the perception of smell.

In normal subjects olfactory evoked cortical potentials are seen with latencies of 150 m/sec and 350 m/sec. To evaluate the site of potential generators in central nervous pathways olfactory evoked cortical potentials were registered in patients with well-defined lesions of the central nervous system and the maxillary nerve. Absence of the first potential was seen in lesions of the brain-stem and the maxillary nerve. Absence of the second potential was observed in lesions of the frontal cerebral medulla and the temporal cortex. No olfactory potentials were found in circumscribed lesions of the postcentral cortex. Sites of the lesions were controlled by computed tomography. Appearance of the two potentials is correlated to normal perception of smell which is established by combined olfactory and trigeminal nerve function.

Brain Diseases↗
PubMed1996

Role of galectin-1 in the developing mouse olfactory system.

Primary sensory olfactory neurons reside in a neuroepithelium lining the nasal cavity and project topographically onto the surface of the olfactory bulb, a rostral extension of the telencephalon. Galectin-1, a bivalent galactose-binding vertebrate lectin, is expressed in the developing rodent olfactory system. In the present study, the mouse olfactory neuron cell line 4.4.2 was used to examine the role of galectin-l in neurite outgrowth in vitro. Recombinant galectin-l has neurite outgrowth-promoting activity when used as a substrate for 4.4.2 cells. When either galectin-1 or lactose was added to the culture media, the neurite outgrowth-promoting activity was abolished. These results demonstrate that galectin-1 can modulate neurite growth in vitro. The in vivo role of galectin-1 was investigated by examining the topographical organization of the olfactory pathway in mice carrying a null mutation for galectin-1. Using Dolichos biflorus agglutinin as a convenient histochemical marker of a subpopulation of primary sensory olfactory neurons which project topographically to the dorsomedial olfactory bulb, we show an aberrant topography of olfactory axons in the null mutants. A subset of primary sensory olfactory axons failed to project to their correct target sites in the caudal olfactory bulb. These data indicate that galectin-1 is involved in the growth and/or guidance of primary sensory olfactory axons between the nasal cavity and the olfactory bulb. This is the first demonstration that a lectin has neurite outgrowth-promoting activity and plays a role in neuronal pathfinding in the mammalian nervous system.

Animals↗
PubMed2005

Information processing in the mammalian olfactory system.

Recently, modern neuroscience has made considerable progress in understanding how the brain perceives, discriminates, and recognizes odorant molecules. This growing knowledge took over when the sense of smell was no longer considered only as a matter for poetry or the perfume industry. Over the last decades, chemical senses captured the attention of scientists who started to investigate the different stages of olfactory pathways. Distinct fields such as genetic, biochemistry, cellular biology, neurophysiology, and behavior have contributed to provide a picture of how odor information is processed in the olfactory system as it moves from the periphery to higher areas of the brain. So far, the combination of these approaches has been most effective at the cellular level, but there are already signs, and even greater hope, that the same is gradually happening at the systems level. This review summarizes the current ideas concerning the cellular mechanisms and organizational strategies used by the olfactory system to process olfactory information. We present findings that exemplified the high degree of olfactory plasticity, with special emphasis on the first central relay of the olfactory system. Recent observations supporting the necessity of such plasticity for adult brain functions are also discussed. Due to space constraints, this review focuses mainly on the olfactory systems of vertebrates, and primarily those of mammals.

Animals↗
PubMed2004

Importance of olfactory and vomeronasal systems for male sexual function.

Chemosensory cues stimulate male sexual arousal and behavior. The main olfactory system has an important role in attracting males to estrous females, and the vomeronasal receptors are important for activating accessory olfactory pathways that engage mating behavior in a sexually dimorphic manner. The gonadotropin releasing hormone (GnRH) neurons like the vomeronasal organ (VNO) neurons take their origin in the olfactory placode and migrate to the basal forebrain along pathfinder axons that take their origin in the developing VNO. The maturation of both systems is synchronized in time such that the early postnatal testosterone surge masculinizes the VNO neural relay en route to the medio preoptic area (MPOA). Although VNO slices and VNO receptor neurons in culture respond to volatile odors, in vivo electrophysiological recordings at the first relay in the accessory olfactory bulb (AOB) are silent until the male makes active nuzzling investigations of the female. The VNO neurons may therefore respond to volatiles that are transported into the organ on carrier peptides that themselves may play a part in receptor activation. In the context of modern molecular phylogenetic studies, it is becoming less likely that pheromones acting via the VNO have any part to play in human sexual behavior, but the possibility exists for conserved VNO genes influencing human reproduction via fertilization.

Animals↗