Circuit properties generating gamma oscillations in a network model of the olfactory bulb

The study of the neural basis of olfaction is important both for understanding the sense of smell and for understanding the mechanisms of neural computation. In the olfactory bulb (OB), the spatial patterning of both sensory inputs and synaptic interactions is crucial for processing odor information, although this patterning alone is not sufficient. Recent studies have suggested that representations of odor may already be distributed and dynamic in the first olfactory relay. The growing evidence demonstrating a functional role for the temporal structure of bulbar neuronal activity supports this assumption. However, the detailed mechanisms underlying this temporal structure have never been thoroughly studied. Our study focused on gamma (40-100 Hz) network oscillations in the mammalian OB, which is a form of temporal patterning in bulbar activity elicited by olfactory stimuli. We used computational modeling combined with electrophysiological recordings to investigate the basic synaptic organization necessary and sufficient to generate sustained gamma rhythms. We found that features of gamma oscillations obtained in vitro were identical to those of a model based on lateral inhibition as the coupling modality (i.e., low irregular firing rate and high oscillation stability). In contrast, they differed substantially from those of a model based on lateral excitatory coupling (i.e., high regular firing rate and instable oscillations). Therefore we could precisely tune the oscillation frequency by changing the kinetics of inhibitory events supporting the lateral inhibition. Moreover, gradually decreasing GABAergic synaptic transmission decreased the degree of relay neuron synchronization in response to sensory inputs, both theoretically and experimentally. Thus we have shown that lateral inhibition provides a mechanism by which the dynamic processing of odor information might be finely tuned within the OB circuit.     

Relationships between odor-elicited oscillations in the salamander olfactory epithelium and olfactory bulb

Oscillations in neuronal population activity, or the synchronous neuronal spiking that underlies them, are thought to play a functional role in sensory processing in the CNS. In the olfactory system, stimulus-induced oscillations are observed both in central processing areas and in the peripheral receptor epithelium. To examine the relationship between these peripheral and central oscillations, we recorded local field potentials simultaneously from the olfactory epithelium and olfactory bulb in tiger salamanders (Ambystoma tigrinum). Stimulus-induced oscillations recorded at these two sites were matched in frequency and slowed concurrently over the time course of the response, suggesting that the oscillations share a common source or are modulated together. Both the power and duration of oscillations increased over a range of amyl acetate concentrations from 2.5 x 10(-2) to 1 x 10(-1) dilution of saturated vapor, but peak frequency was not affected. The frequency of the oscillation did vary with different odorant compounds in both olfactory epithelium and bulb (OE and OB): amyl acetate, ethyl fenchol and d-carvone elicited oscillations of significantly different frequencies, and there was no difference in OE and OB oscillation frequencies. No change in the power or frequency of OE oscillations was observed after sectioning the olfactory nerve, indicating that the OE oscillations have a peripheral source. Finally, application of 1.0 and 10 microM tetrodotoxin to the epithelium blocked OE oscillations in a dose-dependent and reversible manner, suggesting that peripheral olfactory oscillations are related to receptor neuron spiking.     

Properties of reciprocal synapses in the mouse accessory olfactory bulb.

Reciprocal dendrodendritic synapses between mitral and granule cells in the accessory olfactory bulb have been implicated in a specialized form of olfactory learning in mice, in which a female forms a memory to the pheromonal signal of the male that mates with her. Relatively little is known, however, about the mechanism of synaptic transmission at the reciprocal synapses. We analyzed synaptic currents generated in accessory olfactory bulb mitral cells in slice preparations with the patch-clamp technique in nystatin-perforated whole-cell configuration. A brief (5-20-ms) depolarizing voltage step from -70 to 0 mV applied to a single mitral cell evoked GABA(A) receptor-mediated inhibitory postsynaptic currents. The inhibitory postsynaptic currents persisted in the presence of tetrodotoxin, indicating that the inhibitory postsynaptic current in mitral cells can be elicited through purely dendritic interactions. The inhibitory postsynaptic currents were greatly enhanced by washout of extracellular Mg(2+). In Mg(2+)-free solution, the N-methyl-D-aspartate (NMDA) receptor antagonist D,L-2-amino-5-phosphonovaleric acid greatly reduced the inhibitory postsynaptic currents, whereas the non-NMDA receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-(1H,4H)-dione (CNQX) slightly reduced them.These data demonstrate that NMDA receptors play an important role in the generation of dendrodendritic inhibition in mitral cells of the mouse accessory olfactory bulb.     

Alterations in axons and synapses of olfactory cortex following olfactory bulb lesions in newborn rats

Summary The olfactory cortex of rats is being studied at various survival times following deafferentating olfactory bulb ablation on the day of birth. The neonatal axons and synaptic terminals undergo rapid, flocculent degeneration and fragmentation. Most are not electron-dense and therefore probably not argyrophilic at this particular age of the lesion. The degeneration and removal of debris is far more rapid than in adults, yielding a markedly enlarged extracellular space with a relative absence of glia at the vacated postsynaptic thickenings. Denervated postsynaptic thickenings become occupied by neuronal and nonneuronal profiles and profiles of uncertain origin, singly or in various combinations, or the sites may remain partially vacant. One or more axons with synaptic vesicles often aggregated at the site are commonly involved. Certain terminals form contacts on progressively greater lengths of the thickening until typical synaptic contacts predominate by 14 days survival. The results suggest a competitive reinnervation process and provide a fine structural explanation for the events leading to alterations in this pathway following neonatal deafferentation.This project was supported in part by NIH Research Grants DE 04942, awarded by the National Institute of Dental Research, and Grants NS 09678 and NS 04053 from the National Institute of Neurological and Communicative Disorders and Stroke, PHS/DHEWDr. Westrum is also an affiliate of the Child Development and Mental Retardation Center, University of Washington     

Long-term enhancement of synchronized oscillations by adrenergic receptor activation in the olfactory bulb

The noradrenergic system is widely thought to be important for associative learning in the olfactory system through actions in the first processing structure, the main olfactory bulb (MOB). Here, we used extracellular local field potential (LFP) and patch-clamp recordings in rat MOB slices to examine norepinephrine (NE)-induced long-term changes in circuit properties that might underlie learning. During responses to patterned olfactory nerve stimulation mimicking the breathing cycle, NE induced a long-term increase in gamma frequency (30–70 Hz) synchronized oscillations. The enhancement persisted long after washout of NE (70 min), depended on the combined actions of NE and neuronal stimulation, and seemed to be caused by enhanced excitatory drive on the mitral/granule cell network that underlies rapid gamma oscillations. The last effect, increased excitation, was manifested as an increase in evoked long-lasting depolarizations (LLDs) in mitral cells. From a functional perspective, the observed long-term cellular and network changes could promote associative learning by amplifying odor-specific signals.     

Intraglomerular dendritic link connected by gap junctions and chemical synapses in the mouse main olfactory bulb: electron microscopic serial section analyses

Glomeruli of the main olfactory bulb are considered to serve as functional units in processing the olfactory information. Thus the fine tuning of the output level from each glomerulus is important to the information processing in the olfactory system. The interactions among neuronal elements in glomeruli might be one of main mechanisms regulating this output level. In the mouse main olfactory bulb neuronal connections via chemical synapses and gap junction in glomeruli were analyzed by the serial electron microscopical reconstruction. Gap junctions were encountered between diverse types of dendritic processes, between mitral/tufted cell dendrites, between mitral/tufted cell dendrites and periglomerular cell dendrites and between mitral/tufted cell dendrites and dendrites of some interneurons different from periglomerular cells. Then these morphological observations indicate that we must consider both direct coupling between mitral/tufted cells via gap junctions and indirect coupling between mitral/tufted cells via intervening interneuronal processes. One of gap junction-forming processes presynaptic in asymmetrical synapses was traced back to the soma of its origin located in the glomerular layer, which was thus identified as an external tufted cell. However, interestingly, it showed apparently different ultrastructural features from other external tufted cells located at the border between the glomerular and external plexiform layers; the latter resemble so-called mitral/tufted cells located in the external plexiform and mitral cell layers. Then external tufted cells were assumed to be heterogeneous in their ultrastructural features. We occasionally encountered several dendrites connected by gap junctions, which furthermore made chemical synapses with each other and with other surrounding processes. Thus both chemical synapses and gap junctions interconnect complexly various processes in the glomerulus, where the local circuit among intermingled olfactory nerves, mitral/tufted cell dendrites and interneuron dendrites is far more complex than previously schematized.     

A primary culture system of rat olfactory bulb forming many synapses similar to intact ones and spontaneously generating synchronous intracellular calcium oscillations

Previously, several studies attempting to analyze olfactory functions using dissociated culture systems of the olfactory bulb (OB) have been reported. Reciprocal dendrodendritic synapses between secondary neurons (mitral/tufted cells) and interneurons (periglomerular/granule cells) are considered to play the most important role in signal processing in the OB. However, it is unclear whether these reciprocal synapses are formed in vitro in the same way as they are in the intact OB. Thus, we synaptologically investigated the nature of cultured OB neurons. These neurons from embryonic rats were classified into four groups based on the size of their somata and their glutamic acid decarboxylase (GAD) immunoreactivity. At 14 days in vitro, most of the neurons synchronously showed spontaneous intracellular Ca2+ oscillations that were reversibly inhibited by application of D-APV and CNQX. Moreover, the frequency of the oscillations decreased and their amplitude became larger following application of bicuculline. These results suggest functional glutamatergic synaptic coupling and inhibitory GABAergic synaptic modulation. Immunocytochemical staining revealed many dot-like products (puncta) that were immunoreactive to GAD as well as to synaptophysin surrounding the cultured neurons. These results strongly indicate the presence of GABAergic synapses. The existence of synaptic contacts in OB neuron cultures was also confirmed by electron microscopy. Two types of synapses, symmetrical and asymmetrical, were morphologically recognizable. Moreover, we could also identify peculiar synapses resembling the in vivo reciprocal dendrodendritic synapses. The use of these primary culture systems will facilitate the elucidation of mechanisms underlying olfactory functions.     

Excitatory and inhibitory interactions in the olfactory bulb involving dendrodendritic synapses between mitral cells and granular cells

Zusammenfassung Die Größe der maximalen Anstiegsgeschwindigkeit des Ventrikeldruckesdp/dt _max kann durch 2 Grundmechanismen verändert werden. Erstens durch die Faservordehnung und zweitens durch die Änderung der Kontraktionsform der Herzmuskelfaser selbst. Letztere bezeichnet man als Kontraktilität. Durch Kontraktilitätsänderungen kann man vergleichsweise größere Änderungen vondp/dt _max erreichen als durch Variation hämodynamischer Bedingungen. Die Herzfrequenz hat einen relativ geringen Einfluß auf die Kontraktilität.Wird das Maximum der Druckanstiegsgeschwindigkeit in der isovolumetrischen Phase der Ventrikelkontraktion erreicht, so beeinflussen der enddiastolische Ventrikeldruck und der coronare Perfusionsdruck die Höhe vondp/dt _max. Ein erhöhter coronarer Perfusionsdruck steigert zusätzlich die Empfindlichkeit gegenüber Veränderungen des enddiastolischen Druckes. Dies kommt in einer Versteilerung der ventrikulären Funktionskurven zum Ausdruck.Unterhalb eines diastolischen Aortendruckes von 60 mm Hg liegt das Maximum der Druckanstiegsgeschwindigkeit im statistischen Mittel in der Austreibungsphase. Unter Infusion von Isoprenalin wirddp/dt _max ebenfalls nach Öffnung der Aortenklappe erreicht. Der Zeitpunkt vondp/dt _max ist mit der Öffnung der Aortenklappe nicht identisch. Hoher diastolischer Aortendruck, geringe Herzfrequenz und Propranolol verschieben dieses Maximum zeitlich vor die Öffnung der Aortenklappe. Aortendrucksenkung und Isoprenalinzufuhr verschieben dieses Maximum in die Austreibungsphase. Der coronare Perfusionsdruck und der enddiastolische Ventrikeldruck sind ohne Einfluß auf die Lage vondp/dt _max im Herzcyclus.Anspannungszeit und die Zeit vom Beginn der Ventrikelkontraktion bis zum Maximum der Druckanstiegsgeschwindigkeit (t–dp/dt) verhalten sich unterschiedlich. Veränderungen der enddiastolischen Faserspannung, des coronaren Perfusionsdruckes und des diastolischen Aortendruckes sind ohne Einfluß auf die Zeitt–dp/dt. Der diastolische Aortendruck bestimmt allein den Zeitpunkt der Öffnung der Aortenklappe und damit der Anspannungszeit. Bei einer Kontraktilitätsänderung durch Propranolol, Isoprenalin oder Variation der Herzfrequenz verändert sich die Zeitt–dp/dt, in der das Maximum der Druckanstiegsgeschwindigkeit erreicht wird. Erhöht man die Kontraktilität, so steigtdp/dt _max an, und gleichzeitig verkürzt sich die Zeit bis zum Maximum. Senkt man die Kontraktilität, so verlängert sich die Zeitt–dp/dt, und die Größe vondp/dt _max nimmt ab. Auf Grund dieser Befunde erscheint es möglich, Veränderungen desdp/dt _max aufzuteilen in solche, die durch eine Veränderung der Kontraktilität, und in solche, die durch eine Veränderung hämodynamischer Bedingungen bewirkt werden.     

Early X-irradiation of rats--II. Effect on granule cells and their dendrodendritic synapses in the olfactory bulb

Low, repeated doses of X-rays from a Co60 source were used to impair the development of the granule cells and their dendritic terminals in the olfactory bulb, and the resulting effect was studied under light and electron microscopes at 9 days of age. Irradiation of rats from embryonic day 18 (in utero) to postnatal day 5 resulted, among others, in maldevelopment of the (internal) granule cell and external plexiform layers. This was accompanied by a decrease in the number and the density of the granule cells, and the remaining granule cells contained less ribosomes, regardless of their position within the layer. This implies that both supposed subtypes of granule cells were effected. In the external plexiform layer, a reduced number of mature dendrodendritic synapses and signs of harmed granule gemmules were observed. The results suggest that intrauterinal plus postnatal irradiation with low, repeated doses of X-rays may be an effective tool impairing the development of prenatally forming neurons.     

Synaptic organization of the olfactory bulb based on chemical coding of neurons

Olfaction is one of the chemical senses in both vertebrate and invertebrate animals essential for a variety of social behaviors. Recent molecular biological and physiological studies using optical recording have indicated elaborate mechanisms in the main olfactory bulb for processing input from olfactory receptor neurons and control of output to higher centers in the brain. The current challenge is to identify a structural basis for understanding such elaborate molecular and functional organization. Immunocytochemistry and other advanced technologies have enabled us to label bulbar neurons selectively, and they have shown that the olfactory bulb has much greater heterogeneity in chemical and structural neuronal organization and in synaptic connectivity than previously believed. This review describes the structural aspects of the main olfactory bulb of rats and summarizes the findings for its synaptic organization based on chemical coding of neurons. Current uncertainties and issues that need to be clarified in the future are also discussed.     

Strong coupling between pyramidal cell activity and network oscillations in the olfactory cortex

Oscillatory activity is a prominent characteristic of electrophysiological recordings in the olfactory system and has been proposed to play a key role in encoding olfactory representations. Studies in several systems have shown that some aspects of information coding involve characteristics that intertwine spikes and fast oscillations (in the beta and gamma range) of local field potentials (LFP). In the insect olfactory system, it has been proposed that oscillatory activity could provide a temporal link between cells. Following previous data, we have proposed that gamma band oscillations in mammals could subserve a gating function for the transfer of information between the olfactory bulb (OB) and the anterior piriform cortex (aPC), which are functionally coupled. In this study, we used an electrophysiological approach to investigate the temporal relationship between LFP gamma oscillations and single-unit activity by simultaneously recording LFP and single unit discharges in the rat aPC during odor evoked activity. Our data showed that mean spike discharges and gamma oscillatory bursts were synchronized with the same respiratory cycle epoch (around the inspiration/expiration transition). Temporal correlations between spikes and LFP revealed that cortical cell spikes were tightly phase-coupled with the peak of gamma oscillations and that this phase-coupling was not odor-dependent. Our results suggest that gamma oscillation may act as a temporal filter. Oscillatory phase-coupled spikes in the OB could act in increasing the probability of spike emission in the aPC cell during a narrow time-window, explaining the tight phase-coupling observed in the aPC. The role of spike-LFP phase-coupling as a binding function between odor features is discussed.     

Taming time in the olfactory bulb

The biophysics of an unusual synaptic arrangement within the olfactory bulb suggests a way in which rapidly inactivating potassium channels could modulate the timing of oscillations that underlie odor recognition.     

Changes in electrical activity of rabbit olfactory bulb and cortex to conditioned odor stimulation

Rabbits with chronically implanted electrodes in olfactory bulb and cortex were classically conditioned to give an increase in relative frequency of sniffing to odor stimuli (CS+) reinforced with mild electric shock. Electroencephalographic high-frequency (35-85 Hz) bursts were recorded from an ensemble of nine bulbar depth electrodes and a second ensemble of 50 cortical surface electrodes. The olfactory cortex responded to the CS+ with sustained elevation of burst amplitude even though the olfactory bulb, from which it receives its primary centripetal input, underwent a marked decline in burst amplitude during the same time period. The amplitude reduction was not spatially uniform: The burst of the bulbar region that declined most in amplitude had the greatest phase lag with respect to the bulbar ensemble average burst. These effects were learning related because they did not occur for CS+ trials at the beginning of conditioning or for unreinforced control trials at any time.     

Expression of connexin 57 in the olfactory epithelium and olfactory bulb

In the visual system, deletion of connexin 57 (Cx57) reduces gap junction coupling among horizontal cells and results in smaller receptive fields. To explore potential functions of Cx57 in olfaction, in situ hybridization and immunohistochemistry methods were used to investigate expression of Cx57 in the olfactory epithelium and olfactory bulb. Hybridization signal was stronger in the olfactory epithelial layer compared to the connective tissue underneath. Within the sensory epithelial layer, hybridization signal was visible in sublayers containing cell bodies of basal cells and olfactory neurons but not evident at the apical sublayer comprising cell bodies of sustentacular cells. These Cx57 positive cells were clustered into small groups to form different patterns in the olfactory epithelium. However, individual patterns did not associate with specific regions of olfactory turbinates or specific olfactory receptor zones. Patched distribution of hybridization positive cells was also observed in the olfactory bulb and accessory olfactory bulb in layers where granule cells, mitral cells, and juxtaglomerular cells reside. Immunostaining was observed in the cell types described above but the intensity was weaker than that in the retina. This study has provided anatomical basis for future studies on the function of Cx57 in the olfactory system.     

Dendrodendritic inhibition and simulated odor responses in a detailed olfactory bulb network model

In the olfactory bulb, both the spatial distribution and the temporal structure of neuronal activity appear to be important for processing odor information, but it is currently impossible to measure both of these simultaneously with high resolution and in all layers of the bulb. We have developed a biologically realistic model of the mammalian olfactory bulb, incorporating the mitral and granule cells and the dendrodendritic synapses between them, which allows us to observe the network behavior in detail. The cell models were based on previously published work. The attributes of the synapses were obtained from the literature. The pattern of synaptic connections was based on the limited experimental data in the literature on the statistics of connections between neurons in the bulb. The results of simulation experiments with electrical stimulation agree closely in most details with published experimental data. This gives confidence that the model is capturing features of network interactions in the real olfactory bulb. The model predicts that the time course of dendrodendritic inhibition is dependent on the network connectivity as well as on the intrinsic parameters of the synapses. In response to simulated odor stimulation, strongly activated mitral cells tend to suppress neighboring cells, the mitral cells readily synchronize their firing, and increasing the stimulus intensity increases the degree of synchronization. Preliminary experiments suggest that slow temporal changes in the degree of synchronization are more useful in distinguishing between very similar odorants than is the spatial distribution of mean firing rate.