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The effects of local inhibition of N-methyl-D-aspartate and AMPA/kainate receptors in the accessory olfactory bulb on the formation of an olfactory memory in mice.

Female mice form a memory to the pheromones of the mating male, during a critical period after mating. Previous experiments have shown that this memory is located in the accessory olfactory bulb and most likely involves changes at the mitral/granule cell reciprocal synapse. Memory formation can be prevented by local infusions of a non-selective ionotropic glutamate receptor antagonist. However, selective antagonism of N-methyl-D-aspartate receptors during the critical period did not disrupt the memory. The present study shows that memory formation is not prevented by local infusions of the AMPA/kainate receptor antagonist 6,7-dinitroquinoxaline-2,3-dione. Thus, it appears that the formation of a memory to the mating male can occur via both N-methyl-D-aspartate receptor-dependent and independent mechanisms. Far from preventing memory formation, local infusions of 6,7-dinitroquinoxaline-2,3-dione resulted in the formation of a non-selective "memory" to unfamiliar male pheromones. In addition, local infusions of 6,7-dinitroquinoxaline-2,3-dione during pheromonal exposure alone resulted in memory formation without mating having occurred. These results suggest that activity via N-methyl-D-aspartate receptors might be more effective in promoting the synaptic changes underlying memory formation, than activity via AMPA/kainate receptors.

2-Amino-5-phosphonovalerate↗

Electrophysiological evidence for a topographical projection of the nasal mucosa onto the olfactory bulb of the frog.

Three olfactory nerve branches respectively subserving either a medial, an intermediate, or a lateral region of the dorsal olfactory receptor sheet of the bullfrog Rana catesbeiana were electrically stimulated with bipolar platinum hook electrodes. Extracellular single unit responses from 93 second-order cells in different regions of the olfactory bulb were recorded with metal-filled glass micropipets. The excitatory responsiveness of each unit to the stimulation of each of the three nerve branches (response profile) was determined. Some units were sensitive to stimulation of each of the three nerve branches, thus suggesting a wide projection from the entire receptor sheet. On the other hand, other units were more selective. Of this latter group, units in the lateral bulb were excited by nerve branches subserving the more lateral regions of the receptor sheet; units in the medial bulb were excited by the nerve branches subserving the more medial regions of the receptor sheet. These data provide electrophysiological evidence for a topographical projection of the olfactory receptor sheet onto the olfactory bulb, and further suggest that the projections onto different bulbar cells vary in degree of localization.

Animals↗

Entorhinal cortex stimulation modulates amygdala and piriform cortex responses to olfactory bulb inputs in the rat.

The rodent olfactory bulb sends direct projections to the piriform cortex and to two structures intimately implicated in memory processes, the entorhinal cortex and the amygdala. The piriform cortex has monosynaptic projections with the amygdala and the piriform cortex and is therefore in a position to modulate olfactory input either directly in the piriform cortex, or via the amygdala. In order to investigate this hypothesis, field potential signals induced in anesthetized rats by electrical stimulation of the olfactory bulb or the entorhinal cortex were recorded simultaneously in the piriform cortex (anterior part and posterior part) and the amygdala (basolateral nucleus and cortical nucleus). Single-site paired-pulse stimulation was used to assess the time courses of short-term inhibition and facilitation in each recording site in response to electrical stimulation of the olfactory bulb and entorhinal cortex. Paired-pulse stimulation of the olfactory bulb induced homosynaptic inhibition for short interpulse interpulse intervals (20-30 ms) in all the recording sites, with a significantly lower degree of inhibition in the anterior piriform cortex than in the other structures. At longer intervals (40-80 ms), paired-pulse facilitation was observed in all the structures. Paired-pulse stimulation of the entorhinal cortex mainly resulted in inhibition for the shortest interval duration (20 ms) in anterior piriform cortex, posterior piriform cortex and amygdala basolateral but not cortical nucleus. Double-site paired-pulse stimulation was then applied to determine if stimulation of the entorhinal cortex can modulate responses to olfactory bulb stimulation. For short interpulse intervals (20 ms) heterosynaptic inhibition was observed in anterior piriform cortex, posterior piriform cortex and amygdala basolateral but not cortical nucleus. The level of inhibition was greater in the basolateral nucleus than in the other structures. Taken together these data suggest that the entorhinal cortex exerts a main inhibitory effect on the olfactory input via the amygdala basolateral nucleus and to a lesser extent the piriform cortex. The potential role of these effects on the processing of olfactory information is discussed.

Amygdala↗

Microglial activation in the developing rat olfactory bulb.

The development of the olfactory bulb, the primary central relay of the olfactory system, is characterized by a striking susceptibility to alterations in the amount of afferent input. For example, blocking airflow through one half of the nasal cavity during early life results in a number of dramatic changes in the bulb, including increased cell death. Previous studies reveal high levels of microglia in the olfactory bulb. Microglia function as phagocytes, aid in synaptogenesis, and produce important trophic and cytotoxic factors. In response to a number of tissue perturbations, microglia undergo an activation process that includes, among other changes, the up-regulation of complement receptor 3. Interestingly, a previous study reported that naris closure had no effect on microglia in the bulb; however, the research did not distinguish the functional activation state of microglia. We further examined the role of microglia in the normally developing and olfactory-deprived rat bulb using immunohistochemical detection of complement receptor 3 as a measure of microglial activation. Expression of the receptor in the bulb is relatively high during postnatal development, in particular when compared to levels in cortical regions caudal to the olfactory bulb. In addition, naris closure performed on the day after birth (but not after the first postnatal month) increases levels of the receptor in an age and laminar-dependent fashion. The presence of an inducible pool of activated microglia in the olfactory bulb may be important for normal development and contribute to the plethora of changes seen after early olfactory deprivation.

Animals↗

Neonatal and adult neurogenesis provide two distinct populations of newborn neurons to the mouse olfactory bulb.

In mammals, the olfactory bulb (OB) constitutes one of two regions of the postnatal brain with continuous neurogenesis throughout life. Despite intense explorations of neuronal replacement in the adult OB, little is known about the mechanisms that operate at earlier postnatal stages. This question is particularly pertinent, because the majority of local interneurons are born in the neonate, when olfaction controls vital functions. Here, we analyzed the recruitment of newborn cells to the granule cell (GC) layer (GCL) and found that the postnatal mouse OB is supplied with two spatiotemporally distinct populations of newborn interneurons. Early born [postnatal day 3 (P3) to P7] GCs constitute a threefold larger population compared with those generated later (P14-P60), and some of them are produced locally within the OB itself. Newborn interneurons generated at P3-P7 were predominantly targeted to the external edge of the GCL, whereas newly generated cells were positioned deeper in older mice. Additionally, although approximately 50% of adult newborn cells were eliminated within a few weeks of reaching the OB, almost the entire population of early born GCs survived until adulthood. Importantly, early olfactory experience specifically modifies the number of newborn GCs in neonates but leaves unaltered the amount of neurons generated during adulthood. Together, these results demonstrate that early postnatal neurogenesis endows the neonate bulbar circuit with newborn GCs that differ morphologically and functionally from those produced in the adult.

Age Factors↗

The distribution of axon collaterals from the olfactory bulb and the nucleus of the horizontal limb of the diagonal band to the olfactory cortex, demonstrated by double retrograde labeling techniques.

Three different pairs of double retrograde axonal tracers have been used to study the distribution of axon collaterals from individual cells in the olfactory bulb and the nucleus of the horizontal limb of the diagonal band: (1) horseradish peroxidase (HRP) and tritiated apo-HRP (3H-HRP), (2) HRP and 125I-wheat germ agglutinin (I-WGA), and (3) the fluorochromes true blue (TB) and bisbenzimide (BB) or nuclear yellow (NY). With each combination of tracers, paired injections were made into different parts of the olfactory system, and the olfactory bulb and the nucleus of the diagonal band were examined for the presence and arrangement of cells labeled with one or both retrograde tracers. In the olfactory bulb both single and double retrogradely labeled mitral cells were found following injections in disparate parts of the olfactory cortex. Furthermore, no consistent pattern was found in the distribution of single- or double-labeled cells in the olfactory bulb; that is, the distribution of cells labeled from one area of the cortex was not consistently different from the distribution of cells labeled from other parts of the cortex. Therefore, it was concluded that individual mitral cells project to widely spaced parts of the olfactory cortex, and that there is no apparent correspondence between the location of a given cell in the olfactory bulb and the distribution of its axon in the cortex. In contrast to this, cells in the nucleus of the horizontal limb of the diagonal band were only rarely double-labeled from nonoverlapping injections into the olfactory cortex or olfactory bulb, although overlapping injections produced a high proportion of double-labeled cells. Cells which were single-labeled from different injection sites were extensively intermixed within the nucleus. Therefore, in this case it was concluded that individual cells projects to relatively restricted areas, although there was again no apparent correspondence between the position of a cell in the nucleus and the terminal field of its axon.

Animals↗

Functional synapse formation between rat olfactory receptor neurons and olfactory bulb neurons in vitro.

To explore the mechanism of the synapse formation between olfactory receptor neurons and neurons in the olfactory bulb, blocks of olfactory epithelium and slices of olfactory bulb were cocultured by a modified slice culture method. After 4 days in culture, neuron specific fibers from the olfactory epithelium block extending to the olfactory bulb slice were observed. After approximately 20 days in culture, application of forskolin, which induces excitatory responses in olfactory receptor neurons, to the epithelium induced inward current responses in olfactory bulb neurons under the voltage clamp conditions, indicating that functional synapses between olfactory receptor neurons and olfactory bulb neurons had been established in vitro conditions.

Animals↗

Effect of intervening lesions on amino acid distributions in rat olfactory cortex and olfactory bulb.

Levels of the proposed neurotransmitter amino acids glutamate, aspartate, gamma-aminobutyric acid (GABA), and glycine were measured within the layered structures of the olfactory bulb and olfactory cortex following unilateral transections of the lateral olfactory tract or of virtually all fiber tracts of the olfactory peduncle. Distributions of the amino acids on both lesion and control sides were examined and compared by means of a mapping procedure. The results suggest: 1) Glutamate and aspartate are specifically associated with mitral (and presumably also tufted) cell axons and terminals in the piriform cortex. The distribution of aspartate in the olfactory bulb is further suggestive of a specific association of aspartate with mitral cell dendrites and somata. 2) Glutamate might be specifically associated with some centrifugal fibers traveling to the olfactory bulb in or near the anterior commissure. 3) GABA might be specifically related to some certrifugal fibers to the olfactory bulb in addition to its prominent association with granule cells of the bulb. 4) Glycine is unlikely to play a prominent neurotransmitter role in either the olfactory bulb or olfactory cortex.

Amino Acids↗

Modulation of inhibition in a model of olfactory bulb reduces overlap in the neural representation of olfactory stimuli.

In a neural model of olfactory bulb processing, we demonstrate the putative role of the modulation of two types of inhibition, inspired by electrophysiological data on the effect of acetylcholine and noradrenaline on olfactory bulb synaptic transmission. Feedback regulation of modulation based on bulbar activity serves to 'normalize' the activity of output neurons in response to different levels of input activities. This mechanism also decreases the overlap between pairs of output patterns (Mitral cell activities), enhancing the discrimination between overlapping olfactory input patterns. The effect of the modulation at the two levels of interneurons is complementary: while an increase in periglomerular inhibition decreases the number of responding output neurons, a decrease in granule cell inhibition increases the firing frequencies of these neurons.

Computer Simulation↗

Developmental expression of the 3-fucosyl-N-acetyl-lactosamine/CD15 epitope by an olfactory receptor cell subpopulation and in the olfactory bulb of the rat.

In the olfactory epithelium a subpopulation of receptor cells were CD15 (3-fucosyl-N-acetyl-lactosamine)-positive at their somata and their processes. Cytomorphology and location implied their mature character. The labeled neurons were associated with olfactory cilia and showed a unique distribution within the olfactory epithelium and a remarkable numerical change during postnatal ontogeny. The axons of the marked olfactory receptor cells possibly play a role in guiding other axons to their appropriate targets in the olfactory bulb. In the olfactory bulb immunoreactivity was predominantly found in the glomerular and in the external plexiform layer and showed an age-related shift from the superficial to the deep layers.

Animals↗

Functional topography of connections linking mirror-symmetric maps in the mouse olfactory bulb.

In rodents, each main olfactory bulb contains two mirror-symmetric glomerular maps, a feature not found in the initial topographic maps of other sensory systems. Targeting tracer injections to identified glomeruli revealed that isofunctional odor columns-translaminar assemblies connected to a given glomerulus-were specifically and reciprocally interconnected through a mutually inhibitory circuit with exquisite topographic specificity. Thus, instead of containing two mirror-symmetric maps, we propose that the olfactory bulb contains a single integrated map in which isofunctional odor columns are connected through an intrabulbar link, analogous to the specific horizontal connections linking iso-orientation columns in primary visual cortex.

Animals↗

Dynamical mechanisms of odor processing in olfactory bulb mitral cells.

In the olfactory system, the contribution of dynamical properties such as neuronal oscillations and spike synchronization to the representation of odor stimuli is a matter of substantial debate. While relatively simple computational models have sufficed to guide current research in large-scale network dynamics, less attention has been paid to modeling the membrane dynamics in bulbar neurons that may be equally essential to sensory processing. We here present a reduced, conductance-based compartmental model of olfactory bulb mitral cells that exhibits the complex dynamical properties observed in these neurons. Specifically, model neurons exhibit intrinsic subthreshold oscillations with voltage-dependent frequencies that shape the timing of stimulus-evoked action potentials. These oscillations rely on a persistent sodium conductance, an inactivating potassium conductance, and a calcium-dependent potassium conductance and are reset via inhibitory input such as that delivered by periglomerular cell shunt inhibition. Mitral cells fire bursts, or clusters, of spikes when continuously stimulated. Burst properties depend critically on multiple currents, but a progressive deinactivation of I(A) over the course of a burst is an important regulator of burst termination. Each of these complex properties exhibits appropriate dynamics and pharmacology as determined by electrophysiological studies. Additionally, we propose that a second, inconsistently observed form of infrathreshold bistability in mitral cells may derive from the activation of ATP-activated potassium currents responding to hypoxic conditions. We discuss the integration of these cellular properties in the larger context of olfactory bulb network operations.

Algorithms↗

Detecting activity in olfactory bulb glomeruli with astrocyte recording.

In the olfactory bulb, axons of olfactory sensory neurons (OSNs) expressing the same olfactory receptor converge on specific glomeruli. These afferents form axodendritic synapses with mitral/tufted and periglomerular cell dendrites, whereas the dendrites of mitral/tufted cells and periglomerular interneurons form dendrodendritic synapses. The two types of intraglomerular synapses appear to be spatially isolated in subcompartments delineated by astrocyte processes. Because each astrocyte sends processes into a single glomerulus, we used astrocyte recording as an intraglomerular detector of neuronal activity. In glomerular astrocytes, a single shock in the olfactory nerve layer evoked a prolonged inward current, the major part of which was attributable to a barium-sensitive potassium current. The K+ current closely reflected the time course of depolarization of mitral/tufted cells, indicating that K+ accumulation mainly reflects the activity of mitral/tufted cells. The astrocyte K+ current was dependent on AMPA and NMDA receptors in mitral/tufted cells as well as on a previously undescribed metabotropic glutamate receptor 1 component. Block of the K+ current with barium unmasked a synaptic glutamate transporter current. Perhaps surprisingly, the transporter current had components caused by glutamate released at both olfactory nerve terminals and mitral/tufted cell dendrites. The time course of the transporter currents suggested that rapid synchronous glutamate release at OSN terminals triggers asynchronous glutamate release from mitral/tufted cells. Glomerular astrocyte recording provides a sensitive means to examine functional compartmentalization within and between olfactory bulb glomeruli.

Animals↗

Apoptosis following peripheral sensory deafferentation in the olfactory bulb of adult zebrafish.

Removal of the olfactory organ in adult zebrafish results in a significant decrease in volume of the ipsilateral olfactory bulb. The current study investigated the potential role of apoptosis in this phenomenon. It was hypothesized that cells in the adult olfactory bulb normally undergo minimal apoptosis and that apoptosis increases upon removal of sensory stimulation. By using both the terminal transferase-mediated deoxyuridine nick-end labeling method and bis-benzimide labeling, the current study showed that, in the normal adult olfactory bulb, cells exhibiting apoptotic profiles were scarce and were localized to the outer layers of the bulb. However, in deafferented animals, there was a significant increase in the number of apoptotic cells. The apoptotic response occurred in two phases and was confined to the rostral half of the bulb. The first phase of cell death peaked at 1 hour postsurgery. These apoptotic profiles appeared to be primarily nonneuronal in nature, in that they exhibited no immunohistochemical labeling to the neuron-specific protein Hu. The second phase of cell death peaked at 24 hours and declined to normal levels by 1 week. At the 24 hour time point, only a fraction of the apoptotic cells was neuronal in nature. Thus, apoptosis of nonneuronal and neuronal elements accounts for at least part of the deafferentation-induced volume decrease in the zebrafish olfactory bulb. This model of anterograde transneuronal degeneration will be useful in elucidating the afferent signals involved in survival and maintenance of mature brain neurons.

Animals↗

Glutamic acid decarboxylase activity of the olfactory bulb in male rats is influenced by olfactory stimuli and hormonal status.

Changes is glutamate decarboxylase activity (GAD) in the main (MOB) and accessory (AOB) olfactory bulbs were determined at 11:00 hr and 17:00 hr in intact odor deprived (IOD) male rats exposed to female olfactory stimuli, and in castrated odor deprived males (COD) either injected with testosterone (T), exposed to female pheromone or injected with T and exposed to pheromone. Grouping IOD males by female olfactory stimulus and time of the day. MOB and AOB-GAD activity changed in the morning and not in the afternoon. In COD males, T injection induced an increased response in MOB-GAD activity 24 hr later either in the morning or in the afternoon, while no changes were seen at 11:00 or 17:00 hr exposure to female odor. In the AOB of COD males, both T administration or exposure to female pheromone, only induced an increase of enzyme activity in the morning. The association of T and female pheromone, decreased morning GAD activity both in the MOB and in the AOB compared with the values of COD males treated with T. In the afternoon this association had no effect compared with MOB, AOB-GAD activity of COD males injected with testosterone. These results indicate that hormonal and olfactory inputs in IOD and COD males are effective in changing olfactory bulb-GAD activity mainly in the morning.

Animals↗

Calcium influx through NMDA receptors directly evokes GABA release in olfactory bulb granule cells.

Recurrent inhibition in olfactory bulb mitral cells is mediated via reciprocal dendrodendritic synapses with granule cells. Although GABAergic granule cells express both NMDA and non-NMDA glutamate receptors, dendrodendritic inhibition (DDI) relies on the activation of NMDA receptors. Using whole-cell recordings from rat olfactory bulb slices, we now show that olfactory NMDA receptors have a dual role; they depolarize granule cell spines, and they provide a source of calcium that can evoke GABA exocytosis. We demonstrate that exogenous NMDA can trigger GABA release after blockade of voltage-dependent calcium channels (VDCCs) with Cd. We also find that postsynaptic depolarization alone can evoke GABA release via a separate mechanism that relies on calcium influx through Cd-sensitive VDCCs. By selectively manipulating postsynaptic responses in granule cells with high-K or low-Na extracellular solutions, we show that endogenous glutamate can elicit GABA release via both NMDA receptor- and VDCC-dependent pathways. Finally, we find that blockade of Na channels in granule cells with tetrodotoxin enhances DDI, presumably by reducing the depolarization of granule cells during DDI and thereby increasing the driving force for Ca entry through NMDA receptors. These results provide evidence of a novel mechanism for evoked transmitter release that depends on Ca influx through ionotropic receptors and provides a new potential site for synaptic plasticity in the olfactory bulb.

2-Amino-5-phosphonovalerate↗

Comparison of olfactory bulb projections in pigeons and turtles.

The projection targets of the olfactory bulb in pigeons and turtles were investigated using autoradiographic techniques. Despite the relatively smaller size of the olfactory bulbs in pigeons, the projection targets of the olfactory bulb are very similar to those in turtles. In both pigeons and turtles, the olfactory bulb projects to the entire rostrocaudal extent of a portion of the dorsolateral telencephalon (which is here recognized as the pyriform cortex in both birds and reptiles) and to portions of the medial telencephalic wall including the medial septal region. In addition, a projection to the olfactory tubercle of the ventral telencephalon is clearly present in turtles and also appears to be present in pigeons. Pigeons and turtles do differ significantly, however, in the extent of the projection to the amygdaloid region. In turtles, olfactory bulb input encompasses the entire mediolateral and rostrocaudal extent of the amygdaloid region, while in pigeons the input is restricted to a small dorsomedial portion of the amygdala termed nucleus taeniae of the archistriatum. The present results suggest that the olfactory bulb projections in birds are generally similar to those in reptiles, with the exception that secondary olfactory bulb projections to the amygdala may be much reduced in birds compared to those in reptiles. The functional significance of the reduction in olfactory input to the amygdala is presently uncertain.

Animals↗