Odor representations in the rat olfactory bulb change smoothly with morphing stimuli

Many species of mammals are very good at categorizing odors. One model for how this is achieved involves the formation of "attractor" states in the olfactory processing pathway, which converge to stable representations for the odor. We analyzed the responses of rat olfactory bulb mitral/tufted (M/T) cells using stimuli "morphing" from one odor to another through intermediate mixtures. We then developed a phenomenological model for the representation of odors and mixtures by M/T cells and show that >80% of odorant responses to different concentrations and mixtures can be expressed in terms of smoothly summing responses to air and the two pure odorants. Furthermore, the model successfully predicts M/T cell responses to odor mixtures when respiration dependence is eliminated. Thus, odor mixtures are represented in the bulb through summation of components, rather than distinct attractor states. We suggest that our olfactory coding model captures many aspects of single and mixed odor representation in M/T cells.     

Representation of natural stimuli in the rodent main olfactory bulb

Natural odorants are complex mixtures of diverse chemical compounds. Monomolecular odorants are represented in the main olfactory bulb by distinct spatial patterns of activated glomeruli. However, it remains unclear how individual compounds contribute to population representations of natural stimuli, which appear to be unexpectedly sparse. We combined gas chromatography and intrinsic signal imaging to visualize glomerular responses to natural stimuli and their fractionated components. While whole stimuli activated up to 20 visible glomeruli, each fractionated component activated only one or few glomeruli, and most glomeruli were activated by only one component. Thus, responses to complex mixtures reflected activation by multiple components, with each contributing only a small part of the overall representation. We conclude that the population response to a complex stimulus is largely the sum of the responses to its individual components, and activation of an individual glomerulus independently signals the presence of a specific component.     

The source of spontaneous activity in the main olfactory bulb of the rat

Introduction

In vivo, most neurons in the main olfactory bulb exhibit robust spontaneous activity. This paper tests the hypothesis that spontaneous activity in olfactory receptor neurons drives much of the spontaneous activity in mitral and tufted cells via excitatory synapses.

Methods

Single units were recorded in vivo from the main olfactory bulb of a rat before, during, and after application of lidocaine to the olfactory nerve. The effect of lidocaine on the conduction of action potentials from the olfactory epithelium to the olfactory bulb was assessed by electrically stimulating the olfactory nerve rostral to the application site and monitoring the field potential evoked in the bulb.

Results

Lidocaine caused a significant decrease in the amplitude of the olfactory nerve evoked field potential that was recorded in the olfactory bulb. By contrast, the lidocaine block did not significantly alter the spontaneous activity of single units in the bulb, nor did it alter the field potential evoked by electrical stimulation of the lateral olfactory tract. Lidocaine block also did not change the temporal patters of action potential or their synchronization with respiration.

Conclusions

Spontaneous activity in neurons of the main olfactory bulb is not driven mainly by activity in olfactory receptor neurons despite the extensive convergence onto mitral and tufted cells. These results suggest that spontaneous activity of mitral and tufted is either an inherent property of these cells or is driven by centrifugal inputs to the bulb.     

Functional activity of the rat olfactory bulb related to different durations of odor exposure

We have studied the effects of different odor exposure durationson the functional activity of the rat olfactory bulb by usingthe 2-deoxyglucose (2-DG) method. This technique brings outodor-specific patterns of labeling in the glomerular layer ofthe bulb. In a first set of experiments, rats designated ascontrols were submitted to two stimulation conditions with cyclohexanonefor 40 min following 2-DG injection: group C-5—alternatestimulation (5 min odor–5 mm pure air); group C-20—continuousstimulation (20 mm odor-20 min pure air). In a second set ofexperiments, rats designated as ‘adapted’ were exposedfor 3 h to cyclohexanone before the 2-DG injection; then, threeconditions of stimulation were tested with the same odor duringthe post-injection period; group Adap-5—alternate stimulation;group Adap-20—continuous stimulation; group Adap-0—stimulationwith pure air only. In the two control groups, the total numbersof labeled glomerular foci were similar; however, group C-20showed a significant increase in the number of ‘heavily’labeled foci. This result suggests that under conditions of20 min continuous odor exposure, receptor cells do not adaptrapidly and totally. Results from the adapted groups showedthat only the group Adap-20 presented significant changes infunctional bulbar activity. An important decrease in the totalnumber of labeled glomerular foci and an absence of ‘heavily’labeled foci were noted in all rats of this group; their patternsof 2-DG uptake were greatly reduced in both complexity and contrast.These results highlight the importance of olfactory receptoradaptation under conditions of long-duration odor exposure.Fast recovery of receptor sensitivity when the stimulation isinterrupted is also suggested.     

Studies on feminine sexual behavior in the male rat: Influence of olfactory stimuli

The aim of this study was to determine if the display of lordosis behavior in the male rat could be influenced by the olfactory environment. Unexperienced adult male rats were orchidectomized (ORCH). They were primed with 75 μg estradiol benzoate and 1 mg progesterone was injected at an interval of 39 hr following long-term (LT = 3 weeks) or short-term (SHT = 8 hr 30 min) exposure to the odor of male or female urine. For 10 min they were placed in the presence of a “stimulus” male of proven sexual vigor 9 hr 30 min ± 1 hr after progesterone injection. Both LT and SHT exposure to the odor of male urine caused a significant increase in the number of ORCH rats which showed lordosis response to male mounts compared to either the ORCH rats exposed to the odor of female urine or to the controls. Following complete olfactory bulb removal (COBR), no difference was observed in the occurrence of lordosis behavior between the ORCH rats whether or not exposed to the odor of urine. For the ORCH-COBR rats exposed to male urine the proportion of animals responding to mounts did not differ from that of their nonbulbectomized counterparts. In comparing the effects of COBR vs anterior olfactory bulb removal (AOBR) lordosis behavior occurred more frequently in COBR than in AOBR-ORCH rats. The lordosis quotient (LQ) was not affected by exposure to the odor of male urine in the nonbulbectomized ORCH rats. In contrast, it appeared to be higher in both COBR and AOBR animals than in their nonbulbectomized counterparts. The olfactory bulbs were then concluded to inhibit the display of lordosis behavior in the male rat. It was also thought that the olfactory stimuli originating from male urine were capable of releasing the hypothalamic structures involved in the control of lordosis behavior of the male rat from an olfactory inhibitory influence.     

Pregnenolone binding sites in the rat olfactory bulb

High concentrations of pregnenolone and its sulfate have been found in several areas of rat and human brain and seem to be controlled by local mechanisms. In the present experiments we have demonstrated pregnenolone binding sites in the cytosolic fraction of the rat olfactory bulb. The pregnenolone binding component showed a Kd = 2.34 +/- 0.66 x 10(-7) M and Nmax = 7.25 +/- 1.20 pmol/mg protein. Pregnenolone, pregnenolone sulfate and 17OH-pregnenolone competed equally for the binding sites while other steroids were less competitive. Protease and trypsin inhibited binding by 48 and 60% respectively. Sucrose density gradient analysis showed a minor peak at 4.6 s and a major one at 3.6 s. After gel filtration chromatography the pregnenolone binding component appeared as 2 peaks corresponding to molecular weights of approximately 150 and 220 kDa. Heating at 60 degrees C increased binding by 150%. These results indicate that the olfactory bulb pregnenolone binding component is complex in nature. Rat plasma also bound pregnenolone. Plasma binding sites could be partially differentiated from those in the olfactory bulb on the basis of susceptibility to lipoprotein lipase, effect of heating and mobility during polyacrylamide gel electrophoresis.     

Axon mis-targeting in the olfactory bulb during regeneration of olfactory neuroepithelium

During development, primary olfactory axons typically grow to their topographically correct target zone without extensive remodelling. Similarly, in adults, new axons arising from the normal turnover of sensory neurons essentially project to their target without error. In the present study we have examined axon targeting in the olfactory pathway following extensive chemical ablation of the olfactory neuroepithelium in the P2-tau:LacZ line of mice. These mice express LacZ in the P2 subpopulation of primary olfactory neurons whose axons target topographically fixed glomeruli on the medial and lateral surfaces of the olfactory bulb. Intraperitoneal injections of dichlobenil selectively destroyed the sensory neuroepithelium of the nasal cavity without direct physical insult to the olfactory neuron pathway. Primary olfactory neurons regenerated and LacZ staining revealed the trajectory of the P2 axons. Rather than project solely to their topographically appropriate glomeruli, the regenerating P2 axons now terminated in numerous inappropriate glomeruli which were widely dispersed over the olfactory bulb. While these errors in targeting were refined over time, there was still considerable mis-targeting after four months of regeneration.     

Glutamate spillover mediates excitatory transmission in the rat olfactory bulb.

In the CNS, glutamate typically mediates excitatory transmission via local actions at synaptic contacts. In the olfactory bulb, mitral cell dendrites release glutamate at synapses formed only onto the dendrites of inhibitory granule cells. Here, I show excitatory transmission mediated solely by transmitter spillover between mitral cells in olfactory bulb slices. Dendritic glutamate release from individual mitral cells causes self-excitation via local activation of N-methyl-D-aspartate (NMDA) receptors. Paired recordings reveal that glutamate release from one cell generates NMDA receptor-mediated responses in neighboring mitral cells that are enhanced by blockade of glutamate uptake. Furthermore, spillover generates spontaneous NMDA receptor-mediated population responses. This simultaneous activation of neighboring mitral cells by a diffuse action of glutamate provides a mechanism for synchronizing olfactory principal cells.     

The analysis of response similarity in single neurons of the goldfish olfactory bulb using amino-acids as odor stimuli

The response of goldfish olfactory bulb neurons to a range ofamino acid olfactory stimuli was recorded. The results showthat comparisons of response patterns at a single concentrationare inadequate to describe the similarity of response of thesystem to pairs of substances because response patterns changedwith concentration. For some pairs of stimuli, the similaritybetween response patterns was consistent at different concentrationsdespite the changes seen in the responses themselves. In thesecases the similarity of response appeared to reflect the degreeof similarity in chemical structure of the stimuli. Preliminaryevidence is presented that differences in response to differentamino acids are not simply due to differences in stimulatoryeffectiveness for a single receptor process.     

Electrophysiological responses of olfactory bulb neurons to odour stimuli in the frog. A comparison with receptor cells

Extracellular recordings were performed from olfactory bulbneurons in the frog. The odour stimuli were the same as thosepreviously used for studying the receptor cells in the sameanimal species and were delivered at similar concentrations(Revial et al., 1982). The general properties of the neuronresponses are presented and discussed with reference to homologousproperties of olfactory receptor cells. The response rates elicitedby different stimuli from the bulbar neurons were found to behighly correlated with those elicited from receptor cells. Theindividual cell selectivity was better in the bulb than in theolfactory epithelium. The olfactory bulb neurons seemed to improvethe discrimination between stimuli (enantiomers) poorly distinguishedby the receptor cells. Reducing odor concentration caused therate of suppressive response to decrease faster than that ofexcitatory ones, suggesting that the manifestations of inhibitoryprocesses in some neurons requires a high level of excitationin others.     

Proteome profile of the mature rat olfactory bulb

Olfactory bulbs (OBs) are one of the few brain areas, which show active neurogenesis and neuronal migration processes in adult rats. We constructed a proteome map of the 21 days old rat OBs and identified total 196 proteins, out of which 76 proteins were not reported earlier from rat brain. This includes 24 neuronal activity‐specific proteins present at high levels, 7 of which are reported for the first time from OBs.     

Histochemical differentiation between nitric oxide synthase-related and -unrelated diaphorase activity in the rat olfactory bulb

The widely used NADPH-diaphorase reaction for demonstrating neuronal nitric oxide synthase is not as specific as previously thought, as it visualizes both a nitric oxide synthase-related activity and a nitric oxide synthase-unrelated diaphorase. In the present study, we used the rat olfactory bulb as a model to characterize the NADPH-diaphorase activity of neuronal nitric oxide synthase histochemically in comparison with neuronal nitric oxide-unrelated diaphorase activity. The NADPH-diaphorase activity of nitric oxide synthase peaked at pH 8 and at Triton X-100 concentrations of 1--2.5%. It was stable in an acidic environment but was reduced in the presence of Triton X-100 and was inactivated by the flavoprotein inhibitor, diphenyleneiodonium. It preferred beta-NADPH as the co-substrate to alpha-NADPH and alpha-NADH. In contrast, nitric oxide synthase-unrelated diaphorase peaked at pH 10, displayed a Triton X-100 optimum at a concentration of 1%, was unstable in an acidic environment and used beta-NADPH, alpha-NADPH and alpha-NADH to similar extents. Differences in the characteristics between neuronal nitric oxide synthase-related and nitric oxide synthase-unrelated NADPH-diaphorase can be used to increase the specificity of the histochemical nitric oxide synthase marker reaction. © Chapman & Hall