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Genetic manipulation of blood group carbohydrates alters development and pathfinding of primary sensory axons of the olfactory systems.

Primary sensory neurons in the vertebrate olfactory systems are characterised by the differential expression of distinct cell surface carbohydrates. We show here that the histo-blood group H carbohydrate is expressed by primary sensory neurons in both the main and accessory olfactory systems while the blood group A carbohydrate is expressed by a subset of vomeronasal neurons in the developing accessory olfactory system. We have used both loss-of-function and gain-of-function approaches to manipulate expression of these carbohydrates in the olfactory system. In null mutant mice lacking the alpha(1,2)fucosyltransferase FUT1, the absence of blood group H carbohydrate resulted in the delayed maturation of the glomerular layer of the main olfactory bulb. In addition, ubiquitous expression of blood group A on olfactory axons in gain-of-function transgenic mice caused mis-routing of axons in the glomerular layer of the main olfactory bulb and led to exuberant growth of vomeronasal axons in the accessory olfactory bulb. These results provide in vivo evidence for a role of specific cell surface carbohydrates during development of the olfactory nerve pathways.

Animals↗

Neural-dural transition at the medial anterior cranial base: an anatomical and histological study with clinical applications.

OBJECT: Few anatomical studies have been focused on the morphological features and microscopic anatomy of the transition from the intracranial space to the medial anterior cranial base. The authors of the current study performed histological analyses to define the structure of the transition from neural foramina to the cranial base (neural-dural transition) at the cribriform plate, particularly as related to cerebrospinal fluid (CSF) fistula formation and surgical intervention in the region. METHODS: The medial anterior cranial base was resected in six cadaveric specimens. Histological methods were used to study the anatomy of the region on the microscopic level. Results of these examinations revealed a multilayered neural-dural transition at the cribriform plate, which consisted of an arachnoid membrane and a potential subarachnoid space as well as dura mater, periosteum, ethmoid bone, and associated layers of submucosa and mucosa of the paranasal air spaces. A subarachnoid space was identified around the olfactory nerves as they exited the neural foramina of the cribriform plates. The dura mater eventually thinned out and became continuous with the periosteum in the ethmoid bone. The dura, arachnoid membrane, and associated potential subarachnoid space were obliterated at a place 1 to 2 mm into the olfactory foramen. The authors present a case of recurrent CSF rhinorrhea successfully treated using a technique of multilayered reconstruction with pericranium, fat, and bone. CONCLUSIONS: The findings provide an anatomical basis for CSF fistula formation in the region of the cribriform plate and help to explain the unusual presentations in patients who have CSF rhinorrhea and meningitis. These results may facilitate the treatment of CSF fistulas, repair of defects in the medial anterior cranial base, and approaches to tumors and other pathological entities in the region.

Adipose Tissue↗

The nature of unilateral neglect in the olfactory sensory system.

This study investigated two major theories of unilateral neglect utilizing the ipsilaterally innervated olfactory sense. The sensory theory states that unilateral neglect is due to a diminished or attenuated sensory input. The representational theory states that unilateral neglect is due to a disordered internal representation, which is not dependent on sensory input. Results of the study revealed that right hemisphere lesion patients with left unilateral neglect failed to respond to their left contralateral nostril on olfactory double simultaneous stimulation, consistent with the representational theory because the left nostril has no direct sensory input to the right hemisphere.

Attention↗

Lineage analysis of the olfactory epithelium using a replication-incompetent retrovirus.

We have analysed the lineage of olfactory receptor neurons using a replication-incompetent retrovirus injected beneath the olfactory epithelium of young rats. There are two major types of clusters of infected cells seen at 5--40 days after infection: (i) horizontal basal cells (HBCs); (ii) variable numbers of globose basal cells (GBCs), and immature and mature sensory neurons. Olfactory nerve lesion increased the frequency of the globose/sensory neuron clusters, as well as the number of cells/cluster, but did not change the number of HBC clusters or cells/cluster. No clusters contained sustentacular cells. These data indicate that, at least in young rats: (i) HBCs are not precursors of olfactory neurons; (ii) there is a lineage path from GBCs to mature neurons; and (iii) sustentacular cells arise from a separate lineage.

Alkaline Phosphatase↗

Immunocytochemical localization of luteinizing hormone-releasing hormone within the olfactory bulb of pigs.

LHRH was immunocytochemically localized within the olfactory bulb of prepubertal (n = 3), ovariectomized (n = 3), and hypophyseal-stalk-transected (HST) female pigs (n = 3). Perikarya of LHRH-immunoreactive neurons of all pigs were sparsely distributed mostly in the rostral half of the olfactory bulb, along the ventromedial and ventrolateral edge of the olfactory nerve layer, or at its interace with the glomerular layer. Processes from these cells and other LHRH containing axons either entered individual glomeruli forming a network within its interior or coursed around glomeruli penetrating into the external granular layers. Additional fibers penetrated into similar regions of the accessory olfactory bulb. Irregularly shaped perikarya were also detected within the internal granular layer of the ventral olfactory bulb, but only in tissue from HST pigs. From analysis of serial sections, there was no evidence of LHRH projections across the olfactory peduncle that connects the olfactory bulb with adjacent brain regions. If olfactory LHRH neurons are involved in reproductive behavior and physiology in the pig, this pathway involves additional unidentified intervening neurons. Endocrine factors probably influence the expression of immunoreactive LHRH in the internal granule layer, since their presence was revealed only in HST pigs.

Animals↗

Development of olfactory glomeruli: temporal and spatial interactions between olfactory receptor axons and mitral cells in opossums and rats.

Mitral cells are the primary output neurons of the vertebrate olfactory bulb and are major recipients of sensory input from the periphery. The morphogenesis of mitral cell dendrites was followed to elucidate their early spatial and temporal interactions with olfactory receptor neurons and glia during the construction of olfactory glomeruli. Monodelphis domestica, a marsupial born at an extremely immature stage, and rats were examined. Mitral cells were retrogradely labeled by application of the lipophilic dye 1,1' dihexadecyl-3,3,3'3'-tetramethylin-docarbocyanine perchlorate (DiI) to the lateral olfactory tract. In double-labeling experiments, olfactory receptor neurons were stained with 3,3' dihexadecyloxacarbocyanine perchlorate (DiO), or olfactory nerve Schwann cells were visualized using S-100 protein immunohistochemistry. Tissue was examined with a confocal laser scanning microscope. Some preparations were subsequently investigated with an electron microscope. In Monodelphis, differentiation of mitral cells starts with an outgrowth of numerous, uniform, and widespread dendrites. As soon as terminals of olfactory receptor axons coalesce into glomerular knots within the presumptive glomerular layer, dendrites of individual mitral cells innervate several adjacent glomeruli where they receive sensory synaptic input. With maturation, supernumerary mitral cell dendrites retract, leaving one primary dendrite bearing a terminal glomerular tuft. Simultaneously, secondary dendrites begin to arise. The formation of glomeruli begins earlier and progresses faster in the rat compared to Monodelphis. Nevertheless, mitral cell differentiation in both species follows a common sequence: overproduction of dendrites, selection of usually one primary apical dendrite, and elimination of supernumerary processes. Since olfactory receptor neurons form synaptic contacts with the widespread mitral cell dendrites, considerable synaptic rearrangement must occur within the olfactory glomeruli during maturation.

Animals↗

Differential expression patterns of GABA transporters (GAT1-3) in the rat olfactory bulb.

The localization of GABA transporters 1-3 (GAT1-3) was investigated in the rat olfactory bulb by using in situ hybridization and immunohistochemistry. In the glomerular and the internal granular layers, GAT1 mRNA was expressed in most of periglomerular and granule cells, which are known to be GABAergic. In addition, we compared GAT1 mRNA expression with that of glutamic acid decarboxylase67 (GAD67) mRNA. The expressions were very similar in these two layers, indicating that GAT1 mRNA is mainly expressed in GABAergic neurons. However, in the external plexiform and the olfactory nerve layers, we observed more GAT1 mRNA-positive cells than GAD67 ones, suggesting that GAT1 mRNA is also expressed in non-GABAergic neurons and in glial cells. GAT3 mRNA expression was observed in small glial-like cells which might be involved in GABAergic neurotransmission throughout the olfactory bulb. This was confirmed by double-immunostaining studies which showed the expression of both GAT3 and glial fibrillary acidic protein (GFAP) mRNAs in astrocytes. Moreover, GAT2 mRNA was expressed only in the ependyma and arachnoid. These findings suggest that the expression patterns of GABA transporters differ with the type of cells in the rat olfactory bulb where GAT1 and GAT3 may play an imporatant role in GABA-mediated transmission, such as lateral inhibition.

Animals↗

Orthodromic field potentials and recurrent inhibition during anoxia in turtle brain.

Recent evidence suggests that suppression of electrical activity, with the likely reduction of energy use, provides a necessary strategy for prolonged anoxic tolerance of turtle brain. The present study was aimed toward defining mechanisms underlying anoxia-induced suppression of electrical activity in turtle brain. Goals were to determine whether effects of anoxia were selective to excitatory or inhibitory processes and to test the hypothesis that enhanced inhibition underlies suppression of orthodromic field potentials in olfactory bulb. Amplitudes of "conditioning" and "test" responses, provoked by pairs of stimulus pulses to the olfactory nerve, were evaluated before, during, and after anoxic insults in the brains of turtles anesthetized with pentobarbital sodium. Suppression of conditioning responses was proportionally greater than that of test responses, indicating that inhibition was decreased and that enhanced inhibitory activity is not responsible for suppression of field potentials in turtle olfactory bulb during anoxia. When normoxia was restored, conditioning responses recovered but test responses remained suppressed. These data confirm that the inhibitory effect of a conditioning pulse on responses to a second pulse of stimulation results from a recurrent circuit and suggest that anoxia-induced changes in field potentials are independent of this recurrent circuit. Enhanced inhibition during the early period of recovery after anoxic insults may provide advantages for restoring energy status and cell physiology by suppressing electrical activity and energy use.

Animals↗

Heterogeneity of antigen expression and lectin labeling on microglial cells in the olfactory bulb of adult rats.

Microglia in different layers of the rat olfactory bulb expressed a variety of membrane antigens except for CD4 (OX-35). Bulb microglial cells bearing complement receptor type 3 (OX-42) were ubiquitous and their immunoreactivity varied considerably in different bulb layers. Although very few in number, labeled microglia in all layers also expressed major histocompatibility complex class I antigen (OX-18), leukocyte common antigen (OX-1) and unknown macrophage antigen (ED-2). The latter was localized in cells distributed almost exclusively in the perivascular spaces. The immunoreactivity of ED-1, an unknown cytoplasmic or lysosomal membrane antigen in macrophages, was localized in labeled microglia which were concentrated mainly in the granule cell layer and periglomerular zone of the bulb. A variable number of microglial cells were stained with OX-6 (major histocompatibility complex class II antigen) and they were located predominantly in the periglomerular zone and at the junction between the granule cell layer and the subependymal layer. Ultrastructural study using GSA I-B4 lectin labeling showed that microglia in different layers of the bulb exhibited similar labeling patterns in their subcellular structures. A remarkable feature was the occurrence of some microglia in the olfactory nerve layer, subependymal layer and granule cell layer adjacent to the subependymal layer in which the cells showed intense lectin labeling at their Golgi apparatus, a feature which was absent in microglia of other bulb layers. Present results showed the regional differences in microglial antigen expressions and lectin labeling within the olfactory bulb. It is therefore suggested that the cells subserve very different specific functions depending on their ambient microenvironments. The heterogeneity of microglial functions in the olfactory bulb may be related to the progressive regeneration and degeneration of its containing neurons.

Animals↗

Regional distribution of protein kinases in normal and odor-deprived mouse olfactory bulbs.

Unilateral naris closure produced dramatic down-regulation of tyrosine hydroxylase (TH) gene expression in periglomerular dopaminergic neurons in the olfactory bulb. To explore molecular mechanisms of TH gene regulation, the present study investigated the regional distribution of protein kinase A (PKAalpha), protein kinase C (PKCalpha), and CaM kinases II (CaMKIIalpha, beta) and IV (CaMKIV) in the normal olfactory bulb and in response to odor deprivation. Strong PKAalpha immunostaining was found in the glomerular, granule cell, external plexiform and olfactory nerve layers. PKCalpha staining was strong in granule cell and external plexiform layers but weak in the glomerular layer. Whereas CaMKIV was primarily found in granule cells, CaMKII was present in the glomerular, external plexiform, mitral cell and granule cell layers. No change in immunoreactivities of these kinases occurred in the olfactory bulb ipsilateral to naris closure. The expression of PKAalpha, PKCalpha and CaMKII, but not CaMKIV, in periglomerular cells suggests that these three kinases may play a role in TH gene regulation in the olfactory bulb. The lack of change in kinase protein levels after naris closure also suggests that any involvement of these kinases in TH gene expression in the olfactory bulb must be through altered kinase activity and not protein levels.

Animals↗

Histochemical and immunocytochemical study of the migration of neurons from the rat olfactory placode.

Immunocytochemical and histochemical methods have been used to describe the neuronal population migrating from the rat olfactory placode and to analyze the spatio-temporal evolution of this neuronal migration during development. Several neuronal markers, such as binding to the lectin Ulex europaeus (UEA I) and the presence of neuron-specific enolase (NSE), olfactory marker protein (OMP), and luteinizing hormone-releasing hormone (LHRH), have been tested in order to determine whether migrating neurons originate from both the medial and the lateral parts of the placode and whether they all express LHRH. Our data show that a large population of differentiated migrating neurons can be identified with an antibody against NSE from the 14th day of gestation and with UEA I one day later. Migrating neurons are closely associated with both the vomeronasal axon fascicles emerging from the medial pit and the olfactory axons originating from the lateral pit. However, the neuron migration from the lateral pit appears to be more discrete than that from the medial pit. No LHRH immunoreactivity has been detected among neurons migrating from the lateral pit. Some neurons accompanying the olfactory axon fascicles exhibit a high level of maturation as shown by their OMP-positivity. Numerous neurons positive for both NSE and UEA I have also been observed within the presumptive olfactory nerve layer in early embryonic stages.

Animals↗

[Recognition of chemical stimuli by guppy, Poecilia reticulata Peters].

An alimentary conditioned reflex to beta-phenylethanol and a subsequent differentiation of cumarine were elaborated in conditions of simultaneous paired choice in guppies. It was shown that beta-phenylethanol in a concentration not over 7 X 10(-6) M is perceived through olfaction. The conditioned reflex persists for a long time: twenty five to fifty days after section of the olfactory nerves (the conditioned reflex is restored after their regeneration) and at least seventy days in intact fishes.

Animals↗

Distribution of vasoactive intestinal polypeptide-like immunoreactivity in the olfactory bulb of the rainbow trout (Salmo gairdneri).

The distribution of vasoactive intestinal polypeptide-like structures in the olfactory bulb of the rainbow trout was studied using an indirect-immunoperoxidase technique. Olfactory fibres were very strongly labelled, whereas the fibres or cell bodies in the remaining strata of the olfactory bulb showed no immunoreactivity. In addition, the olfactory nerve fibres were not immunoreactive for methionine- and leucine-enkephalins, motilin, neuropeptide Y, substance P, cholecystokinin-8 and tyrosine-hydroxylase.

Animals↗

Differential labelling of primary olfactory system subcomponents by SBA (lectin) and NADPH-d histochemistry in the frog Pipa.

SBA and NADPH-d histochemistries allow identification of functionally distinct components of the amphibian primary olfactory system. In Pipa, a secondarily aquatic frog, combination of both methodologies, using alternate sets of histological sections, reveals that, apart from Jacobson's organ, this species has a "water-nose" and an "air-nose". The epithelia occupy separate chambers of the olfactory organ and give rise to olfactory nerve fiber bundles that are identified by the dual staining procedure.

Adaptation, Physiological↗

[The developmental expression of embryonic-neural cell adhesive molecule in human olfactory mucosa].

OBJECTIVE: To discuss the expressional features and physical function of E-NCAM (embryonic- neural cell adhesive molecule) in human fetal olfactory mucosa. METHOD: The immune expressions of E-NCAM in human olfactory mucosa at different gestational stages was detected with the technique of immunohistochemistry. RESULT: The positive expression of E-NCAM began at gestational week 14 (14W), and was localized in the cell bodied, dendrites, axis of olfactory neurons and olfactory nerve fiber bundles in lamina propria. The positive cells increased and the densities of staining of fiber bundles rose with the development of the fetus and reached the peak at 28W and decreased gradually then on. And at each stage the E-NCAM positive cells were localized at middle-lower parts of olfactory mucosa while the sustentacular cells and neurons at the top part were always E-NCAM negative. CONCLUSION: The expression features of E-NCAM are coincident with the plastict transformation of olfactory neurons. The high expression of E-NCAM at early half parts of gestation is helpful to the growth of neurons in differentiation and the down-regulation at late-half parts of gestation is helpful to the stabilization of structure of mature neurons.

Embryo, Mammalian↗