5-Hydroxytryptamine action in the rat olfactory bulb: in vitro electrophysiological patch-clamp recordings of juxtaglomerular and mitral cells.

The olfactory bulb, first relay of olfactory pathways, is densely innervated by serotoninergic centrifugal fibers originating from the raphe nuclei. Although serotonin innervation was reported to be involved in olfactory learning in mammals, the action of this neurotransmitter on its putative cellular targets has been never described through unitary recordings. This lack of data initiated the present study where the effects of 5HT on juxtaglomerular and mitral cells are analyzed using whole-cell recordings on olfactory bulb slices. Serotonin depolarizes 34% of 525 JG cells. A multivariate statistical analysis of juxtaglomerular cells characteristics shows that the serotonin responsive cell group can be individualized regarding their tonic discharge-mode in response to a direct current injection, their lower expression of hyperpolarization-activated cation current and their low membrane capacities. The use of ion channel blockers and ramp voltage protocol indicate that serotoninergic depolarization of juxtaglomerular cells may be due to a nonselective cation current with a reversal potential of -44 mV. Pharmacological tests with serotonin receptor antagonists and agonists reveal that 5HT action on juxtaglomerular cells would be mainly mediated by 5HT2C receptors. In mitral cells, serotonin acts on 49.1% of the 242 tested cells, inducing two types of responses. A first subset of mitral cells (26.8%, n=65) were hyperpolarized by serotonin. This response would be indirect and mediated by action of GABA on GABAA receptors since it was antagonized by bicuculline. The involved GABAergic neurons are hypothesized to be juxtaglomerular and granular cells, on which serotonin would act mainly via 5HT2C and via 5HT2A receptors respectively. The second subset of mitral cells (22.3%, n=54) were directly depolarized by serotonin acting through 5HT2A receptors. Our data on serotonin action on juxtaglomerular cells and mitral cells reveal a part of functional mechanisms whereby serotonin can act on olfactory bulb network. This is expected to enrich the understanding of its determining role in olfactory learning.

Taurine action on mitral cell activity in the frog olfactory bulb in vivo.

Taurine (TAU) is a free amino acid that is particularly abundant in the olfactory bulb. In the frog, TAU is located in the terminations of the primary olfactory axons and in the granular cell layer. TAU action seems to be associated with gamma amino butyric acid (GABA), the main inhibitory neurotransmitter involved in the processing of the sensory signal. The present study was designed to assess the action of TAU in vivo during the olfactory network's stimulation by odors. It was performed by recording the single-unit activity of mitral cells, the main bulbar output neurons. TAU effects were tested on both their spontaneous and odor-induced firing activity. Interactions between TAU and GABA were examined by analyzing TAU effects under the selective blocking action of GABAA or GABAB antagonists. TAU was found to suppress the spontaneous firing of mitral cells, mainly without altering their odor response properties. By testing GABA antagonists, we further show that TAU action is associated with GABAergic inhibitory mechanisms mainly via GABAB receptors. Thus, TAU action clearly reduces background activity in favor of the emergence of the odor-induced activity in the same manner as GABA action does via GABAB receptors. As a conclusion, we propose that, in the frog olfactory bulb, the joint actions of TAU and GABA may favor the processing of the primary sensory information by increasing the signal to noise ratio.

GABA(B) receptor-mediated inhibition of mitral/tufted cell activity in the rat olfactory bulb: a whole-cell patch-clamp study in vitro.

GABA, the major inhibitory neurotransmitter involved in information processing in the olfactory bulb, is hypothesized to act through GABA(B) receptors by depressing primary neurotransmitter release at the level of olfactory nerve axon endings. The present study was designed to analyze GABA(B) receptor-mediated inhibition mechanisms by performing whole-cell patch-clamp recordings of mitral/tufted cell activity in the rat in vitro. To do so, GABA(B) receptor-mediated action was mimicked by baclofen and antagonized by saclofen. Our protocol led us to provide an original description of GABA(B) receptor-mediated inhibition exerted on mitral/tufted cells. First, their spontaneous activity was shown to be drastically abolished by baclofen. Second, their responses to olfactory nerve electrical stimulation were graded by GABA(B) receptor-mediated inhibition. Indeed, this inhibition may be described as inducing effects ranked from a slight increase in response latency to a complete response suppression.Altogether, our results corroborate the hypothesis of a presynaptic extrasynaptic GABA(B) receptor-mediated inhibition influencing mitral/tufted cell olfactory nerve responsivity. However, the involvement of postsynaptic receptors, with different properties or with different anatomical locations, cannot be ruled out, particularly in the control of spontaneous activity. In conclusion, we underline that, in the vertebrate olfactory bulb, GABA(B) receptor-mediated action appears to contribute to make mitral/tufted cell responses more salient by reducing their resting activity.

Sensory information processing in the frog olfactory pathways. Experimental basis for modeling studies.

In the frog, unitary electrophysiological recordings have been extensively used to investigate odor processing along the olfactory pathways. By comparing spontaneous and odor-evoked activities of neuroreceptor, mitral and cortical cells, we have collected fundamental data relating to coding abilities of the three olfactory levels, the olfactory mucosa, the bulb and the cortex. Based on a synthesis of our experimental data related to GABAergic and dopaminergic involvement in the olfactory bulb, this paper aims to match this information with computational data and to discuss some questions on bulbar processing. This paper is also devoted to further analyze original results on coding properties of two functionally evidenced neuron subpopulations in the olfactory cortex. Thus, the assumption according to which some cortical neurons may work as temporal integrators while others as coincidence detectors is presented. Moreover, the pertinence that the neural code may be carried by a single spike with varying latency was demonstrated.

Odor coding properties of frog olfactory cortical neurons.

Until now, in amphibians, response odor properties of primary cortical neurons had never been investigated. Furthermore, very few data on this subject are available in other species. This prompted us to explore the functional properties of olfactory cortical neurons at rest and in response to odors. To achieve this, our experience with odor coding in the first two stages of the frog olfactory system, the olfactory mucosa and the olfactory bulb, led us to use odor stimuli which were chemical compounds with known stimulating properties, delivered to the mucosa in controlled conditions over a wide concentration range. Most of the cortical neurons were found to be very silent at rest, their average spontaneous activity being significantly lower than that of bulb neurons recorded previously in the same conditions. Cortical cells displayed, with all odors combined, 35% excitatory responses and 8% inhibitory responses. The excitatory response rate was similar to that of the bulb, while the inhibitory response rate was about 4.5-fold lower. Interestingly, two functional groups of cortical cells emerged based both on differences in response temporal patterning to odors delivered at increasing concentrations and in qualitative discrimination power. Regarding intensity coding, group 1 cells (53%) displayed "classical" temporal pattern evolution, increase of discharge frequencies and decrease of latency and burst duration, over the concentration range. The responses of group 2 cells (47%) were clearly original, since they consisted of a single spike (or more rarely two spikes) occurring with a strictly reproducible latency at a given concentration and a decreased latency as a function of increasing concentration. The dynamics of cell recruitment in the cortex showed that group 1 cell recruitment mimicked that of mitral cells, group 2 cells being recruited at higher concentrations. The analysis of qualitative discrimination properties of cortical cells regarding the eight-odor set revealed that the discrimination power of group 2 cells was similar to that of mitral cells. By contrast, the qualitative discrimination power of group 1 cells was found to be similar to that of neuroreceptor cells. In conclusion, this pioneer approach leads us to report that olfactory cortical neurons of the frog are responsive to odors and can be clearly divided into two groups based on functional criteria. Group 1 cells, which were relatively selective, poorly discriminating but sensitive, may be mainly devoted to intensity coding. By contrast, group 2 cells, which were not very sensitive but were selective and discriminating, were hypothesized to provide minimal intensity coding and thus to be mainly devoted to qualitative discrimination tasks.

Action of vasopressin on hypoglossal motoneurones of the rat: presynaptic and postsynaptic effects.

The distribution of vasopressin binding sites in the hypoglossal nucleus of newborn rats was determined using autoradiography on film and a radioiodinated vasopressor antagonist. These sites predominated in the ventromedial and dorsal divisions of the nucleus. The effect of vasopressin on hypoglossal neurones was studied in brainstem slices of newborn animals, using the single-electrode voltage-clamp technique. Vasopressin, at 0.1-0.5 microM, generated a sustained inward current in a majority of neurones, an action which was mediated by V1-type receptors. Antidromic activation or morphological characterization of biocytin-labelled neurones indicate that part of the vasopressin-sensitive cells were motoneurones. When synaptic transmission was blocked by perfusing the preparation with a low-calcium/high-magnesium solution, the average vasopressin current decreased by 65%; and following TTX treatment, the peptide current decreased by 55%. In contrast, in a low-calcium solution, i.e., under conditions of reduced synaptic transmission but of increased neuronal excitability, the vasopressin current was not significantly altered. These results may be interpreted by assuming that the action of vasopressin is in part postsynaptic and in part presynaptic, the latter effect probably depending upon action potential propagation. Current-voltage relations suggest that the postsynaptic effect of vasopressin was due to the induction of a non-inactivating inward current, reversing in polarity at around -15 mV. The data raise the possibility that, in young animals, endogenous vasopressin may modulate the activity of hypoglossal motoneurones.

Somatic Acetylcholine Release in Rabbit Nodose Ganglion.

In the rabbit, as in various other species, the presence of a cholinergic vagal afferent contingent has been demonstrated previously using biochemical and immunohistological approaches at the nodose ganglion level, where vagal afferent cell bodies are located. This structure is completely devoid of synaptic contacts. In the present study, somatic acetylcholine release is demonstrated on different types of in vitro rabbit nodose ganglion preparations (fragments of nodose tissue or isolated cell bodies) using chemiluminescent detection. Acetylcholine endogenous content was measured and was shown to be greater in the right nodose ganglion compared to the left. This difference was also observed when spontaneous and potassium chloride-evoked acetylcholine release was measured in extracellular fluid after a 15-min incubation of nodose ganglion fragments. Calcium removal totally blocked this somatic release. A kinetic study of acetylcholine release was also performed by placing the samples (nodose ganglion fragments or isolated cell bodies) directly in front of the photomultiplier, allowing the direct monitoring of (acetylcholine + choline) and choline effluxes. The net acetylcholine release was then deduced by subtraction. Identical kinetics was obtained with the two different nodose ganglion preparations used. This somatic release is calcium-dependent. The occurrence of acetylcholine release at the nodose ganglion level is discussed in comparison with the events occurring in the cholinergic nerve endings. These mechanisms could be implicated in the premodulation of the vagal afferent messages conveyed from the periphery to the central nervous system.

Choline uptake in cholinergic nodose cell bodies.

Isolated living cell bodies were obtained by mechanical and enzymatic dissociation from adult rabbit nodose ganglion followed by separation of fibres and cells using a Percoll gradient. A purification yield of 45% was measured. Based on previous results obtained in whole ganglion and showing the presence of cholinergic cell bodies among the afferent fibres of the vagus nerve, this preparation was used to study choline uptake by neuron cell somata. Cholinergic cells counted after choline acetyltransferase immunohistological staining showed a stained population of 2.9% among the isolated population. Two [3H]choline uptake mechanisms were detected at the cell body level. The first, with Km1 = 7 microM and Vm1 = 200 pmol/h per ganglion is sodium dependent, related to acetylcholine synthesis (43%) and has an IC50 with hemicholinium-3 equal to 50 microM. The second, with Km2 = 54 microM and Vm2 = 2235 pmol/h per ganglion is sodium independent, poorly associated to acetylcholine synthesis (12%) and exhibits an IC50 of 2 microM with hemicholinium-3. Except for their sensitivity to hemicholinium-3, the high and low affinity choline uptake mechanisms observed at the somatic level have, respectively, the same characteristics as the high and low affinity mechanisms described at the synaptic level. Their physiological role, their opposed sensitivity to hemicholinium-3 compared to the synaptic uptake systems and the relation between the somatic high affinity choline transport and an acetylcholine somatic release are discussed.