A Golgi study on the main olfactory bulb in the snake Elaphe quadrivirgata

The intrinsic organization of the main olfactory bulb in the snake was studied using the rapid Golgi method. A distinct laminar structure was recognized. From the periphery inward, the following layers were distinguished: the layer of the olfactory fibers, the olfactory glomeruli, the mitral cells, the deep fiber plexus, the granule cells and the ependymal cells. Olfactory fibers derived from the nasal cavity reached the entire surface of the bulb, forming a dense fiber plexus, then swung deeply and terminated in the olfactory glomeruli which were arranged in 2-4 rows. The mitral cell layer occupied a wide zone and was composed of scattered mitral cells. The mitral cells had 2-9 primary dendrites proceeding externally to terminate in the olfactory glomeruli and 2-4 secondary dendrites extending tangentially in the mitral cell layer to be distributed therein. The axons of the mitral cells travelled deeply and entered the layer of the deep fiber plexus. The deep fiber plexus was the path for the bulbar efferent and afferent fibers and could be traced caudally as the main olfactory tract, up to the anterior olfactory nucleus and vicinity. The granule cell layer was composed of small cells, the granule cells, packed closely with no special arrangement. The granule cells had long processes which extended superficially to be distributed mainly in the mitral cell layer. The ependymal cells were located at the deepest layer forming the wall of the olfactory ventricle and generated a long process which extended towards the surface to terminate in the peripheral portion of the bulb. In the snake bulb, the well-documented external and internal plexiform layers were considered to be included in the wide mitral cell layer. Thus, while several specific structures were observed, the fundamental organization of the main olfactory bulb in the snake seemed to be identical to that of the main olfactory bulb in various other vertebrate species.     

Centrifugal innervation of the mammalian olfactory bulb

Although it has been known for decades that the mammalian olfactory bulb receives a substantial number of centrifugal inputs from other regions of the brain, relatively few data have been available on the function of the centrifugal olfactory system. Knowing the role of the centrifugal projection and how it works is of critical importance to fully understanding olfaction. The centrifugal fibers can be classified into two groups, a group that release neuromodulators, such as noradrenaiine, serotonin, or acetylcholine, and a group originating in the olfactory cortex. Accumulating evidence suggests that centrifugal neuromodulatory inputs are associated with acquisition of odor memory. Because the distribution of the terminals on these fibers is diffuse and widespread, the neuromodulatory inputs must affect diverse subsets of bulbar neurons at the same time. In contrast, knowledge of the role of centrifugal fibers from the olfactory cortical areas is limited. Judging from recent morphological evidence, these fibers may modify the activity of neurons located in sparse and discrete loci in the olfactory bulb. Given the modular organization of the olfactory bulb, centrifugal fibers from the olfactory cortex may help coordinate the activities of restricted subsets of neurons belonging to distinct functional modules in an odor-specific manner. Because the olfactory cortex receives inputs from limbic and neocortical areas in addition to inputs from the bulb, the centrifugal inputs from the cortex can modulate odor processing in the bulb in response to non-olfactory as well as olfactory cues.     

GABAergic basal forebrain afferents innervate selectively GABAergic targets in the main olfactory bulb

In this work we have analyzed the targets of the GABAergic afferents to the main olfactory bulb originating in the basal forebrain of the rat. We combined anterograde tracing of 10 kD biotinylated dextran amine (BDA) injected in the region of the horizontal limb of the diagonal band of Broca that projects to the main olfactory bulb, with immunocytochemical detection of GABA under electron microscopy or vesicular GABA transporter (vGABAt) under confocal fluorescent microscopy. GABAergic afferents were identified as double labeled BDA-GABA boutons. Their targets were identified by their ultrastructure and GABA content. We found that GABAergic afferents from the basal forebrain were distributed all over the bulbar lamination, but were more abundant in the glomerular and inframitral layers (i.e. internal plexiform layer and granule cell layer). The fibers had thick varicosities with abundant mitochondria and large perforated synaptic specializations. They contacted exclusively GABAergic cells, corresponding to type 1 periglomerular cells in the glomerular layer, and to granule cells in inframitral layers. This innervation will synchronize the bulbar inhibition and consequently the response of the principal cells to the olfactory input. The effect of the activation of this pathway will produce a disinhibition of the bulbar principal cells. This facilitation might occur at two separate levels: first in the terminal tufts of mitral and tufted cells via inhibition of type 1 periglomerular cells; second at the level of the firing of the principal cells via inhibition of granule cells. The GABAergic projection from the basal forebrain ends selectively on interneurons, specifically on type 1 periglomerular cells and granule cells, and is likely to control the activity of the olfactory bulb via disinhibition of principal cells. Possible similarities of this pathway with the septo-hippocampal loop are discussed.     

Complementary postsynaptic activity patterns elicited in olfactory bulb by stimulation of mitral/tufted and centrifugal fiber inputs to granule cells

Main olfactory bulb (MOB) granule cells receive spatially segregated glutamatergic synaptic inputs from the dendrites of mitral/tufted cells as well as from the axons of centrifugal fibers (CFFs) originating in olfactory cortical areas. Dendrodendritic synapses from mitral/tufted cells occur on granule cell distal dendrites in the external plexiform layer (EPL), whereas CFFs preferentially target the somata/proximal dendrites of granule cells in the granule cell layer (GCL). In the present study, tract tracing, and recordings of field potentials and voltage-sensitive dye optical signals were used to map activity patterns elicited by activation of these two inputs to granule cells in mouse olfactory bulb slices. Stimulation of the lateral olfactory tract (LOT) produced a negative field potential in the EPL and a positivity in the GCL. CFF stimulation produced field potentials of opposite polarity in the EPL and GCL to those elicited by LOT. LOT-evoked optical signals appeared in the EPL and spread subsequently to deeper layers, whereas CFF-evoked responses appeared in the GCL and then spread superficially. Evoked responses were reduced by N-methyl-d-aspartate (NMDA) receptor antagonists and completely suppressed by AMPA receptor antagonists. Reduction of extracellular Mg(2+) enhanced the strength and spatiotemporal extent of the evoked responses. These and additional findings indicate that LOT- and CFF-evoked field potentials and optical signals reflect postsynaptic activity in granule cells, with moderate NMDA and dominant AMPA receptor components. Taken together, these results demonstrate that LOT and CFF stimulation in MOB slices selectively activate glutamatergic inputs to the distal dendrites versus somata/proximal dendrites of granule cells.     

A Golgi study on the olfactory bulb in the lamprey, Lampetra japonica

The intrinsic organization of the olfactory bulb in the lamprey was studied using the rapid Golgi method. Although not as discrete as in many vertebrates, a laminar organization was recognized. From the periphery inward, the following layers were discernible: the layer of the olfactory fibers, the olfactory glomeruli with the mitral cells, the granule cells, and the ependymal cells. Just beneath the surface of the olfactory bulb, the olfactory fibers extended over the entire bulb forming a dense fiber plexus terminating in the olfactory glomeruli which were arranged in one to two layers internally to the layer of the olfactory fibers. The mitral cells formed no discrete layer and were located mainly around the olfactory glomeruli. The mitral cells in the lamprey were lacking in secondary dendrites, but had two or more primary dendrites which terminated in the olfactory glomeruli. The axons of the mitral cells proceeded inwardly and accumulated diffusely in the granule cell layer which occupied a wide area internally to the layer of the olfactory glomeruli with the mitral cells. The granule cell layer was composed of densely packed small spindle or fusiform axonless cells, the processes of which extended superficially to be distributed in the olfactory glomeruli. At the deepest region of the bulb was a layer of the ependymal cells lining the surface of the olfactory ventricle. The external and internal plexiform layers were not evident. Thus, while the major constituents of the olfactory bulb of the vertebrate could be identified in that of the lamprey, the general laminar organization seemed indiscrete.     

Central olfactory connections in the microsmatic marmoset monkey (Callithrix jacchus)

The mammalian primary olfactory system consists of a set of different telencephalic structures, including paleo-, archi-, periarchi- and mesocortical components. We present the first characterisation of the normal and connectional anatomy of the primary olfactory cortex of the common marmoset, a microsmatic simian species increasingly used in primate research. The centrifugal and centripetal bulbar projections were determined by injections of the anterograde and retrograde tracer wheat germ agglutinin-conjugated horseradish peroxidase and fluorescent dyes into the ipsilateral main olfactory bulb. The efferent projections of the marmoset bulb are organised entirely ipsilaterally and are established via a rudimentary medial olfactory tract and the dominant lateral olfactory tract. Target areas are the anterior olfactory nucleus, the entire prepiriform cortex, ventral tenia tecta, periamygdaloid cortex and the rostral part of the entorhinal cortex. The bulbar axons predominantly terminate in the outer part of layer I. The anterior olfactory nucleus receives a weak additional input within layer II and III, which is not found in macrosmatic rodents. Further anterograde labelling was found in the endopiriform nucleus deep under the prepiriform cortex and within an anterolateral strip of the olfactory tubercle. However, control injections into the olfactory tubercle suggest that the marmoset olfactory tubercle receives a bisynaptic olfactory input only. Retrograde labelling after bulb injections revealed that, except for the olfactory tubercle, all primary olfactory cortices contributed to an ipsilateral bulbopetal feedback projection. Like in rodents, the only bulbopetal projection organised bilaterally in the marmoset is maintained by the anterior olfactory nucleus. With few exceptions, the projections of the marmoset olfactory brain are organised similarly to that of the macaque monkey or those of macrosmatic species.     

Centrifugal regulation of neuronal activity in the olfactory bulb of the waking rabbit as revealed by reversible cryogenic blockade

Summary The influences of centrifugal projections to the olfactory bulb were examined on the bulbar EEG and mitral-tufted cell activity in waking rabbits. Each of 6 rabbits was implanted, under surgical anesthesia, with fine wire electrodes for recording of the EEG and mitral-tufted cell unit activity and for stimulating the lateral olfactory tract. Two cooling probes, for reversible cryogenic blockade, were implanted on either side of the left olfactory peduncle. Records of EEG and unit activity were taken for 200 s before, during and after cooling of the probes to 3 degrees centigrade. Antidromic evoked potentials were used to assess the efficacy of the blockade. During the cryogenic blockade bursts of EEG activity, evoked in the bulb by inspiration through the nose, were augmented in amplitude and reduced in frequency. Mitral-tufted cell unit activity was reduced in rate but was more highly correlated with the phase and amplitude of the EEG bursts. Analysis of individual EEG bursts revealed that the variance in frequency of bulbar activity was significantly reduced in the isolated state. The data demonstrate that oscillatory bursting activity in the olfactory bulb is intrinsically maintained within a relatively fixed frequency range during receptor input and does not depend on centrifugal projections for its electrogenesis. Changes in EEG frequency, amplitude and correlation with unit activity support the hypothesis that centrifugal projections act in part to inhibit mitral-tufted cell output by direct excitation of granule cells. These findings are supported by a theoretical model in which distributed feedback to the granule cells from more central olfactory structures acts to regulate the coherency of bulbar activity.     

Topography of centrifugal acetylcholinesterase-positive fibres in the olfactory bulb of the rat: evidence for original projections in atypical glomeruli

An original pathway of centrifugal acetylcholinesterase-positive fibres is described in the olfactory bulb of the rat. A dense network of positive fibres spreads out superficially at the boundaries of the lateral olfactory tract and the glomerular layer. These labelled fibres converge towards atypical glomerular structures lying close to the classical olfactory glomeruli. The atypical glomeruli are located dorsally at the medial border of the accessory olfactory bulb, in the area previously described as the "modified glomerular complex", and in the ventrolateral bulbar area. They structurally differ from typical glomeruli, as suggested by observations on semithin sections. The ultrastructural distribution of acetylcholinesterases into axonal and dendritic profiles, around and inside atypical glomeruli, is consistent with the hypothesis of centrifugal modulatory influences at this level. This study illustrates several new aspects of morphofunctional heterogeneity in the olfactory system. The glomerular layer of the main olfactory bulb can no longer be considered as morphologically and functionally uniform. Atypical glomeruli located in the mediodorsal and the ventrolateral boundaries of the glomerular layer are characterized by both structural features and an uncommonly high convergence of acetylcholinesterase-positive centrifugal fibres. Such areas might be involved in the processing of specific olfactory signals as demonstrated elsewhere for the "modified glomerular complex".     

Origin and migration of interneurons of the olfactory bulb in the musk shrew,Suncus murinus

The olfactory bulb of the musk shrew, Suncus murinus, is characterized by the presence of various interneurons. Our previous report (Kakuta et al., 2001) demonstrated that positive immunoreactions for calretinin were observed in periglomerular and perinidal cells in the glomerular layer, small ovoid neurons in the external plexiform layer, and granule cells in the granule cell layer of the olfactory bulb in the musk shrew aged 1 to 5 weeks, in addition to calretinin-immunoreactive bipolar cells distributed in the anterior subependymal layer and in each layer of the olfactory bulb. To examine the origin and migration of interneurons of the olfactory bulb, we labeled generated cells by injecting 28-day-old musk shrews with 5-bromo-2'-deoxyuridine (BrdU), and detected the labeled progeny cells that survived after several intervals. BrdU-labeled cells originated in the subependymal layer around the anterior horn of the lateral ventricle, and rostrally migrated in the subependymal layer from the anterior wall of the lateral ventricle into the center of the olfactory bulb, where they radially migrated into the granule cell layer, external plexiform layer, and glomerular layer. It took 2 days to migrate rostrally in the subependymal layer from the anterior lateral ventricle to the center of the olfactory bulb, and 2 to 6 days to migrate radially from the bulbar subependymal layer into the three layers mentioned. The rate of rostralward migration of the labeled cells was estimated to be 38 microm/h, while that of radial migration, 7 to 25 microm/h. The present BrdU-labeling study, together with our previous immunohistochemical study (Kakuta et al., 2001), indicates that anterior subependymal cells differentiate into granule cells in the granule cell layer, into Van Gehuchten cells in the external plexiform layer, and into periglomerular and perinidal cells in the glomerular layer of the olfactory bulb in the musk shrew.     

The olfactory marker protein in the olfactory system of the mouse during development

The olfactory marker protein, a protein specific to the olfactory sensory neurons, has been studied in mouse during embryogenesis and in the postnatal period up to 30 days, with the unlabeled antibody enzyme method of immunohistochemistry. Olfactory neurons, which are morphologically detectable in 10-day-old embryos, do not contain olfactory marker protein. The protein is present in the olfactory neuroepithelium at embryonic day 14 and its appearance coincides with the establishment of sensory synapses in the olfactory bulb. Neurons containing the protein increase in number up to 30 days after birth. At 15 days of embryonic life, immunostaining was observed in sensory axons at the rostral tip of the olfactory bulb, and by embryonic day 17 a plexus of stained fibres has covered the bulbar surface. Between embryonic day 15 and postnatal day 1, olfactory axons have been observed to reach the mitral cell layer. In the vomeronasal system the olfactory marker protein is present at later stages and both the receptors' perikarya and their axons and axon terminals in the accessory olfactory bulb show a lower level of staining than the olfactory system proper.This study of the olfactory marker protein has allowed us to correlate its appearance with significant developmental phenomena.     

Dopaminergic neuromodulation of synaptic transmission between mitral and granule cells in the teleost olfactory bulb

A growing body of evidence suggests that teleosts are important models for the study of neural processing of olfactory information, and the functional role of dopamine (DA), which is a potent neuromodulator endogenous to the mammalian olfactory bulb, has been one of the strongest focuses in this field. However, the cellular mechanisms of dopaminergic neuromodulation in olfactory bulbar neural circuits have not been fully understood. We investigated such mechanisms by using the goldfish, which offers several advantages for analyzing olfactory information processing by electrophysiological methods. First, we found in the olfactory bulb that numerous cell bodies of the dopaminergic neurons are mainly distributed in the mitral cell layer and extend fine processes to the glomerular layer. Next, we made in vitro field potential recordings and showed that synaptic transmissions from mitral to granule cells were suppressed by DA application. DA also increased the paired-pulse ratio, suggesting that the suppression of synaptic transmission is caused by a decrease in presynaptic glutamate release from the mitral cells. Furthermore, DA significantly suppressed the oscillatory activity of the olfactory bulb in response to olfactory stimuli. Although DA suppresses the synaptic inputs from the olfactory nerve to the olfactory bulbar neurons in mammals, this phenomenon was not observed in the goldfish. These findings indicate that suppression of the mitral to granule cell synaptic transmission in the reciprocal synapses plays an important role in the negative regulation of olfactory responsiveness in the goldfish olfactory bulb.     

An experimental study of the projection of the amygdala to the accessory olfactory bulb and its relationship to the concept of a dual olfactory system

Summary The present anatomical findings point to the existence of a separate subdivision of the olfactory system whose connections are quite different from the principal part. The main olfactory bulb has olfactory afferents from the receptors of the general olfactory mucosa, while the accessory bulb has afferents from receptors in the vomeronasal organ. The main bulb projects to the olfactory tubercle and pyriform cortex, while the accessory bulb projects to the amygdala. In turn these areas are further related with the medial forebrain bundle in the case of the pyriform cortex and olfactory tubercle, and with the medial preoptic area and medial hypothalamus in the case of the amygdala. The main and accessory olfactory bulbs are further distinguished by their centrifugal connections, the main bulb receiving fibres from the olfactory tubercle passing through the lateral olfactory tract, and the accessory olfactory bulb receiving fibres from the amygdala through the stria terminalis. The centrifugals to the accessory olfactory bulb resemble those to the main bulb in that both appear to terminate upon granule cells, although further projections to the external plexiform layer or to the periglomerular region have not been demonstrated for the accessory bulb. By virtue of its neural connections the accessory olfactory system is ideally placed to mediate the effects of olfactory stimuli on reproduction.     

Neuropeptide Y in the olfactory system, forebrain and pituitary of the teleost, Clarias batrachus

Distribution of neuropeptide Y (NPY)-like immunoreactivity in the forebrain of catfish Clarias batrachus was examined with immunocytochemistry. Conspicuous immunoreactivity was seen in the olfactory receptor neurons (ORNs), their projections in the olfactory nerve, fascicles of the olfactory nerve layer in the periphery of bulb and in the medial olfactory tracts as they extend to the telencephalic lobes. Ablation of the olfactory organ resulted in loss of immunoreactivity in the olfactory nerve layer of the bulb and also in the fascicles of the medial olfactory tracts. This evidence suggests that NPY may serve as a neurotransmitter in the ORNs and convey chemosensory information to the olfactory bulb, and also to the telencephalon over the extrabulbar projections. In addition, network of beaded immunoreactive fibers was noticed throughout the olfactory bulb, which did not respond to ablation experiment. These fibers may represent centrifugal innervation of the bulb. Strong immunoreactivity was encountered in some ganglion cells of nervus terminalis. Immunoreactive fibers and terminal fields were widely distributed in the telencephalon. Several neurons of nucleus entopeduncularis were moderately immunoreactive; and a small population of neurons in nucleus preopticus periventricularis was also labeled. Immunoreactive terminal fields were particularly conspicuous in the preoptic, the tuberal areas, and the periventricular zone around the third ventricle and inferior lobes. NPY immunoreactive cells and fibers were detected in all the lobes of the pituitary gland. Present results describing the localization of NPY in the forebrain of C. batrachus are in concurrence with the pattern of the immunoreactivity encountered in other teleosts. However, NPY in olfactory system of C. batrachus is a novel feature that suggests a role for the peptide in processing of chemosensory information.     

Neurosteroid modulation of dendrodendritic inhibition in the mouse olfactory bulb

The present report describes neurosteroid modulation of olfactory bulb function by examining the effects of intrabulbar infusion of dehydroepiandrosterone sulfate (DHEAS), a neurohormone discovered in rat brain, on field potentials in the granule cell layer evoked by paired-pulse stimulation of the mouse lateral olfactory tract. Infusion of DHEAS (5 nmol) significantly decreased the test response without affecting the conditioning response. As a consequence, DHEAS selectively potentiated paired-pulse depression, which is believed to be due to granule cell-mediated inhibition of the mitral/tufted cells. The granule-to-mitral/tufted dendrodendritic synapse is GABAergic. Taken together, these results suggest that DHEAS potentiates the GABAergic dendrodendritic inhibition exerted by the granule cells on the mitral/tufted cells.     

Heterogeneous targeting of centrifugal inputs to the glomerular layer of the main olfactory bulb

The centrifugal systems innervating the olfactory bulb are important elements in the functional regulation of the olfactory pathway. In this study, the selective innervation of specific glomeruli by serotonergic, noradrenergic and cholinergic centrifugal axons was analyzed. Thus, the morphology, distribution and density of positive axons were studied in the glomerular layer of the main olfactory bulb of the rat, using serotonin-, serotonin transporter- and dopamine-β-hydroxylase-immunohistochemistry and acetylcholinesterase histochemistry in serial sections. Serotonin-, serotonin transporter-immunostaining and acetylcholinesterase-staining revealed a higher heterogeneity in the glomerular layer of the main olfactory bulb than previously reported. In this sense, four types of glomeruli could be identified according to their serotonergic innervation. The main distinctive feature of these four types of glomeruli was their serotonergic fibre density, although they also differed in their size, morphology and relative position throughout the rostro-caudal main olfactory bulb. In this sense, some specific regions of the glomerular layer were occupied by glomeruli with a particular morphology and a characteristic serotonergic innervation pattern that was consistent from animal to animal. Regarding the cholinergic system, we offer a new subclassification of glomeruli based on the distribution of cholinergic fibres in the glomerular structure. Finally, the serotonergic and cholinergic innervation patterns were compared in the glomerular layer. Sexual differences concerning the density of serotonergic fibres were observed in the atypical glomeruli (characterized by their strong cholinergic innervation). The present report provides new data on the heterogeneity of the centrifugal innervation of the glomerular layer that constitutes the morphological substrate supporting the existence of differential modulatory levels among the entire glomerular population.     

Expression of the dopaminergic phenotype in the olfactory bulb: neither calcitonin gene-related peptide nor olfactory input is necessary.

In the olfactory bulb, expression of tyrosine hydroxylase (TH) in juxtaglomerular neurons is dependent on innervation by the olfactory nerve. The presence of the neuropeptide calcitonin gene-related peptide (CGRP) within the olfactory nerve has led to the hypothesis that CGRP is responsible for regulation of TH expression in the bulbar neurons. On the other hand, other investigators claim that olfactory receptors never produce CGRP and that functional contact with olfactory axons regulates production of TH by bulbar neurons. Two different experimental procedures were used to test whether either CGRP or contact with the olfactory nerve is essential for production of TH by bulbar neurons in vivo. The peptidergic innervation of the olfactory bulb was eliminated either by neonatal capsaicin treatment, or by stereotaxic, electrolytic lesions of the ophthalmic division of the trigeminal nerve. Both of the treatments leave the olfactory innervation of the bulb intact while eliminating the CGRP-immunoreactive fibers in the olfactory nerve and glomeruli. Subsequent immunocytochemistry reveals a normal complement of bulbar TH-immunoreactive juxtaglomerular neurons in the absence of peptidergic innervation. In order to test whether olfactory nerve input is necessary for expression of TH in vivo, the anlage of the olfactory bulb was removed from embryonic (E16) rat pups and transplanted into the anterior chamber. These ectopic olfactory bulbs, although devoid of olfactory nerve input, contain numerous TH-immunoreactive neurons. Thus olfactory nerve input is not necessary for expression of TH in bulbar neurons.     

Distribution of enkephalin-like immunoreactivity in the rat main olfactory bulb

Enkephalin-like immunoreactivity was localized within the main olfactory bulb of the rat using immunohistochemical techniques. These studies utilized well characterized antisera directed to either leu⁵- or met⁵-enkephalin. Specificity was established by absorption of the antisera with either 10 μM synthetic leu⁵- or met⁵-enkephalin.Specific enkephalin-like immunoreactivity was observed within several different cell populations including (1) periglomerular cells, (2) granule cells and their processes within the external plexiform layer and (3) occasional short-axon (horizontal) cells within the granule and external plaxiform layers. The granule cell layer contained the greatest number of immunoreactive cells. Only a limited number of immunoreactive cells were found in both the periglomerular and granule cell layers, suggesting the enkephalin-containing neurons represent a sub-population within each layer.The absence of immunoreactive processes in the periventribular white matter, as well as the morphologies of immunoreactive bulbar neurons, indicates that enkephalin is found exclusively within intrinsic olfactory bulb neurons.     

Gabaergic control of olfactory learning in young rats.

Olfactory learning in young rats correlates with neural plasticity in the olfactory bulb, and involves noradrenergic modulation of reciprocal dendrodendritic synapses between mitral cells and GABAergic granule cells. The purpose of this study was to examine, in vivo, the consequences of manipulating bulbar GABA transmission during training. In the first experiment, postnatal day 11 rat pups were trained in an olfactory associative learning task with citral odor and foot shock as the conditioned and unconditioned stimuli, respectively. The pups received continuous infusion of saline or the GABA(A) receptor agonist muscimol into the olfactory bulbs throughout a 30-min training session. The pups were then tested on postnatal day 12 for a preference for or an aversion to citral odor. Saline-infused control pups developed an aversion to citral odor. The GABA(A) receptor agonist muscimol impaired this aversive learning in a dose-dependent manner. In the second experiment, pups were exposed to the odor for 30 min while receiving continuous intrabulbar infusion of a low or high dose of the GABA(A) receptor antagonist bicuculline, without any other reinforcer. Depending on whether a low (0.2 nmol/bulb) or high (1.0 nmol/bulb) dose of bicuculline was infused, the pups showed a preference or an aversion for citral odor after infusion of low and high doses, respectively. These results indicate that disinhibition of mitral cells in the olfactory bulb is critical for olfactory learning in young rats, and suggest that the degree of disinhibition is an important determinant in acquiring either preference or aversion for the conditioned odor.     

Synaptic connectivity of serotonergic axons in the olfactory glomeruli of the rat olfactory bulb

Although the major mode of transmission for serotonin in the brain is volume transmission, previous anatomical studies have demonstrated that serotonergic axons do form synaptic contacts. The olfactory glomeruli of the olfactory bulb of mammals receive a strong serotonergic innervation from the dorsal and medial raphe nuclei. In the present report, we investigate the synaptic connectivity of these serotonergic axons in the glomerular neuropil of the rat olfactory bulb. Our study shows that serotonergic axons form asymmetrical synaptic contacts on dendrites within the glomerular neuropil. Analyzing the neurochemical nature of the synaptic targets, we have found that 55% of the synapses were on GABA-immunopositive profiles and 45% on GABA-immunonegative profiles. These data indicate that barely half of the contacts were found in GABA-immunonegative profiles and half of the synapses in GABA-positive dendrites belonging to type 1 periglomerular cells. Synaptic contacts from serotonergic axons on dendrites of principal cells cannot be excluded, since some of the GABA-immunonegative postsynaptic profiles contacted by serotonergic axons had the typical ultrastructural features of bulbar principal cell dendrites. Altogether, our results suggest a complex action of the serotonergic system in the modulation of the bulbar circuitry.     

Developmental, tract-tracing and immunohistochemical study of the peripheral olfactory system in a basal vertebrate: insights on Pax6 neurons migrating along the olfactory nerve

The olfactory system represents an excellent model for studying different aspects of the development of the nervous system ranging from neurogenesis to mechanisms of axon growth and guidance. Important findings in this field come from comparative studies. We have analyzed key events in the development of the olfactory system of the shark Scyliorhinus canicula by combining immunohistochemical and tract-tracing methods. We describe for the first time in a cartilaginous fish an early population of pioneer HuC/D-immunoreactive (ir) neurons that seemed to delaminate from the olfactory pit epithelium and migrate toward the telencephalon before the olfactory nerve was identifiable. A distinct, transient cell population, namely the migratory mass, courses later on in apposition to the developing olfactory nerve. It contains olfactory ensheathing glial (GFAP-ir) cells and HuC/D-ir neurons, some of which course toward an extrabulbar region. We also demonstrate that Pax6-ir cells coursing along the developing olfactory pathways in S. canicula are young migrating (HuC/D and DCX-ir) neurons of the migratory mass that do not form part of the terminal nerve pathway. Evidences that these Pax6 neurons originate in the olfactory epithelium are also reported. As Pax6 neurons in the olfactory epithelium show characteristics of olfactory receptor neurons, and migrating Pax6-ir neurons formed transient corridors along the course of olfactory axons at the entrance of the olfactory bulb, we propose that these neurons could play a role as guideposts for axons of olfactory receptor neurons growing toward the olfactory bulb.