Antennocerebral Tract Research Articles

Choline acetyltransferase‐like immunoreactivity in a physiologically distinct subtype of olfactory nonspiking local interneurons in the cockroach (periplaneta americana)

Behavioral and physiological studies have shown that local interneurons are pivotal for processing odor information in the insect antennal lobe. They mediate inhibitory and excitatory interactions between the glomerular pathways and ultimately shape the tuning profile of projection neurons. To identify putative cholinergic local interneurons in the antennal lobe of Periplaneta americana, an antibody raised against the biosynthetic enzyme choline acetyltransferase (ChAT) was applied to individual morphologically and electrophysiologically characterized local interneurons. In nonspiking type IIa1 local interneurons, which were classified in this study, we found ChAT-like immunoreactivity suggesting that they are most likely excitatory. This is a well-defined population of neurons that generates Ca(2+) -driven spikelets upon depolarization and stimulation with odorants, but not Na(+) -driven action potentials, because they lack voltage-activated transient Na(+) currents. The nonspiking type IIa2 and type IIb local interneurons, in which Ca(2+) -driven spikelets were absent, had no ChAT-like immunoreactivity. The GABA-like immunoreactive, spiking type I local interneurons had no ChAT-like immunoreactivity. In addition, we showed that uniglomerular projection neurons with cell bodies located in the ventral portion of the ventrolateral somata group and projections along the inner antennocerebral tract exhibited ChAT-like immunoreactivity. Assigning potential transmitters and neuromodulators to distinct morphological and electrophysiological types of antennal lobe neurons is an important prerequisite for a detailed understanding of odor information processing in insects.

Neuropeptide-Gated Perception of Appetitive Olfactory Inputs in Drosophila Larvae

Understanding how smell or taste translates into behavior remains challenging. We have developed a behavioral paradigm in Drosophila larvae to investigate reception and processing of appetitive olfactory inputs in higher-order olfactory centers. We found that the brief presentation of appetitive odors caused fed larvae to display impulsive feeding of sugar-rich food. Deficiencies in the signaling of neuropeptide F (NPF), the fly counterpart of neuropeptide Y (NPY), blocked appetitive odor-induced feeding by disrupting dopamine (DA)-mediated higher-order olfactory processing. We have identified a small number of appetitive odor-responsive dopaminergic neurons (DL2) whose activation mimics the behavioral effect of appetitive odor stimulation. Both NPF and DL2 neurons project to the secondary olfactory processing center; NPF and its receptor NPFR1 mediate a gating mechanism for reception of olfactory inputs in DL2 neurons. Our findings suggest that eating for reward value is an ancient behavior and that fly larvae are useful for studying neurobiology and the evolution of olfactory reward-driven behavior.

Hydroxyurea Ablation of Mushroom Bodies inDrosophila

Chemical ablation is an effective tool for studying nervous system development and function in Drosophila. Hydroxyurea (HU) inhibits ribonucleotide reductase, blocking DNA synthesis, and killing dividing cells. The specificity of HU ablation is thus dependent on developmental events. In this respect, HU is useful in determining temporal patterns of neuroblast proliferation and the origins of neuronal elements in flies and other insects. In Drosophila, an especially fortuitous time window occurs at the end of embryonic development. For the first 8-12 h after larval hatching, only five neuroblasts are proliferating in each brain hemisphere. Four of these are found in the dorsal protocerebrum and give rise to the intrinsic elements (Kenyon cells [KCs] and glia) of the mushroom bodies (MBs). The remaining single neuroblast has an anterolateral position in the brain and is the progenitor of local interneurons (LocI) in the antennal lobe (AL) and a subset of lateral relay interneurons (RIl) in the inner antennocerebral tract (iACT). Treating newly hatched larvae with HU results in adult flies with KCs and AL interneurons of embryonic origin only. This protocol describes methods for collecting newly hatched Drosophila larvae and treating them with HU.

Regeneration of olfactory afferent axons in the locust brain

AbstractConfocal image of a locust brain 14 days after one antenna had been crushed. Antennal afferents were anterogradely labeled with Neurobiotin (magenta). Aberrantly regenerated olfactory receptor axons have grown through the antennocerebral tract into the mushroom body. Counterstaining: phalloidin (green) and DAPI (blue). The Journal of Comparative Neurology, Volume 520, Number 4, pages 679‐693.

Heterogeneity in dendritic morphology of moth antennal lobe projection neurons

AbstractA population of projection neurons (PNs) in the antennal lobe (AL) integrates sensory information from the antenna and is essential for processing odor information in the insect brain. We examined the anatomy of this neuronal population in the brain of the silkmoth Bombyx mori. Using intracellular dye injection, we labeled a total of 246 PNs and systematically analyzed their morphological features, including the soma position, antennocerebral tract, and number of innervating glomeruli. For example, we analyzed PNs that had somata in the different cell clusters, innervated overlapping but different groups of glomeruli, and ran through different pathways. We also identified glomeruli innervated by PNs using a previously established procedure that first classifies glomeruli into regional groups and then identifies individual glomeruli. We analyzed uniglomerular PNs (75.6% of the total) and found heterogeneity in the dendritic morphology of the PNs that was dependent on the regions and/or the innervating glomeruli. For example, most PNs innervating the macroglomerular complex did not have extraglomerular processes, whereas most PNs innervating ordinary glomeruli did. Moreover, PNs innervating the toroid glomerulus showed heterogeneity in their dendritic morphology. These PNs had dendritic arborization in different areas within the glomerulus. We found that, in some cases, the innervation pattern of the PN dendrite correlated with individual variation in the glomerular organization. These results indicate that PNs are not homogeneous populations, and in some cases morphological heterogeneity in PNs correlated with change in glomerular organization in the silkmoth AL. J. Comp. Neurol. , 2011. © 2011 Wiley‐Liss, Inc.

Heterogeneity in dendritic morphology of moth antennal lobe projection neurons

A population of projection neurons (PNs) in the antennal lobe (AL) integrates sensory information from the antenna and is essential for processing odor information in the insect brain. We examined the anatomy of this neuronal population in the brain of the silkmoth Bombyx mori. Using intracellular dye injection, we labeled a total of 246 PNs and systematically analyzed their morphological features, including the soma position, antennocerebral tract, and number of innervating glomeruli. For example, we analyzed PNs that had somata in the different cell clusters, innervated overlapping but different groups of glomeruli, and ran through different pathways. We also identified glomeruli innervated by PNs using a previously established procedure that first classifies glomeruli into regional groups and then identifies individual glomeruli. We analyzed uniglomerular PNs (75.6% of the total) and found heterogeneity in the dendritic morphology of the PNs that was dependent on the regions and/or the innervating glomeruli. For example, most PNs innervating the macroglomerular complex did not have extraglomerular processes, whereas most PNs innervating ordinary glomeruli did. Moreover, PNs innervating the toroid glomerulus showed heterogeneity in their dendritic morphology. These PNs had dendritic arborization in different areas within the glomerulus. We found that, in some cases, the innervation pattern of the PN dendrite correlated with individual variation in the glomerular organization. These results indicate that PNs are not homogeneous populations, and in some cases morphological heterogeneity in PNs correlated with change in glomerular organization in the silkmoth AL.

Highly Tissue Specific Expression of Sphinx Supports Its Male Courtship Related Role in Drosophila melanogaster

Sphinx is a lineage-specific non-coding RNA gene involved in regulating courtship behavior in Drosophila melanogaster. The 5′ flanking region of the gene is conserved across Drosophila species, with the proximal 300 bp being conserved out to D. virilis and a further 600 bp region being conserved amongst the melanogaster subgroup (D. melanogaster, D. simulans, D. sechellia, D. yakuba, and D. erecta). Using a green fluorescence protein transformation system, we demonstrated that a 253 bp region of the highly conserved segment was sufficient to drive sphinx expression in male accessory gland. GFP signals were also observed in brain, wing hairs and leg bristles. An additional ∼800 bp upstream region was able to enhance expression specifically in proboscis, suggesting the existence of enhancer elements. Using anti-GFP staining, we identified putative sphinx expression signal in the brain antennal lobe and inner antennocerebral tract, suggesting that sphinx might be involved in olfactory neuron mediated regulation of male courtship behavior. Whole genome expression profiling of the sphinx knockout mutation identified significant up-regulated gene categories related to accessory gland protein function and odor perception, suggesting sphinx might be a negative regulator of its target genes.

Parallel Representation of Stimulus Identity and Intensity in a Dual Pathway Model Inspired by the Olfactory System of the Honeybee

The honeybee Apis mellifera has a remarkable ability to detect and locate food sources during foraging, and to associate odor cues with food rewards. In the honeybee’s olfactory system, sensory input is first processed in the antennal lobe (AL) network. Uniglomerular projection neurons (PNs) convey the sensory code from the AL to higher brain regions via two parallel but anatomically distinct pathways, the lateral and the medial antenno-cerebral tract (l- and m-ACT). Neurons innervating either tract show characteristic differences in odor selectivity, concentration dependence, and representation of mixtures. It is still unknown how this differential stimulus representation is achieved within the AL network. In this contribution, we use a computational network model to demonstrate that the experimentally observed features of odor coding in PNs can be reproduced by varying lateral inhibition and gain control in an otherwise unchanged AL network. We show that odor coding in the l-ACT supports detection and accurate identification of weak odor traces at the expense of concentration sensitivity, while odor coding in the m-ACT provides the basis for the computation and following of concentration gradients but provides weaker discrimination power. Both coding strategies are mutually exclusive, which creates a tradeoff between detection accuracy and sensitivity. The development of two parallel systems may thus reflect an evolutionary solution to this problem that enables honeybees to achieve both tasks during bee foraging in their natural environment, and which could inspire the development of artificial chemosensory devices for odor-guided navigation in robots.

Morphological and Physiological Characteristics of the Serotonin-Immunoreactive Neuron in the Antennal Lobe of the Male Oriental Tobacco Budworm, Helicoverpa assulta

We have characterized, by intracellular recording and staining combined with immunocytochemistry, a serotonin-immunoreactive neuron in the central olfactory pathway of the male moth Helicoverpa assulta. The neuron joins the unique category of so-called SI antennal-lobe neurons, previously described in several insect species. In similarity with that originally discovered in the sphinx moth Manduca sexta, the neuron identified here has a large soma located posteriorly in the lateral cell cluster of the antennal lobe and an unbranched neurite projecting into the ipsilateral protocerebrum via the inner antennocerebral tract. After bypassing the central body, the axon crosses the midline and extends through the corresponding antennocerebral tract to the contralateral antennal lobe where it innervates the entire assembly of glomeruli including the male-specific macroglomerular complex. The neuron arborizes into several fine branches in bilateral protocerebral regions anterior to the calyces of the mushroom bodies, particularly on the contralateral side. The physiology of the neuron revealed 2 distinctly different spiking amplitudes, 1 small showing a relatively high spontaneous activity and 1 large showing low activity. The small-amplitude spikes displayed increased frequency when pheromones and plant odors were blown over the antenna. The large-amplitude spikes, which had an unusually long duration, showed no observable responses.

Gamma‐aminobutyric acid (GABA)‐mediated neural connections in the Drosophila antennal lobe

Inhibitory synaptic connections mediated by gamma-aminobutyric acid (GABA) play important roles in the neural computation of the brain. To obtain a detailed overview of the neural connections mediated by GABA signals, we analyzed the distribution of the cells that produce and receive GABA in the Drosophila adult brain. Relatively small numbers of the cells, which form clusters in several areas of the brain, express the GABA synthesis enzyme Gad1. On the other hand, many cells scattered across the brain express ionotropic GABA(A) receptor subunits (Lcch3 and Rdl) and metabotropic GABA(B) receptor subtypes (GABA-B-R1, -2, and -3). To analyze the expression of these genes in distinct identified cell types, we focused on the antennal lobe, where GABAergic neurons play important roles in odor coding. By combining fluorescent in situ hybridization and immunolabeling against GFP expressed with cell-type-specific GAL4 driver strains, we quantified the percentage of the cells that produce or receive GABA for each cell type. GABA was synthesized in the middle antennocerebral tract (mACT) projection neurons and two types of local neurons. Among them, mACT neurons had few presynaptic sites in the antennal lobe, making the local neurons essentially the sole provider of GABA signals there. On the other hand, not only these local neurons but also all types of projection neurons expressed both ionotropic and metabotropic GABA receptors. Thus, even though inhibitory signals are released from only a few, specific types of local neurons, the signals are read by most of the neurons in the antennal lobe neural circuitry.

γ-Aminobutyric acid immunostaining in the antennal lobe of the moth Heliothis virescens and its colocalization with neuropeptides

The antennal lobe is the primary processing center for olfactory information in insects. To understand further the neural circuitry of this brain area, we have investigated the distribution of gamma-aminobutyric acid (GABA) and its colocalization with neuropeptides in the antennal lobe of the noctuid moth Heliothis virescens. Immunocytochemical experiments with an antiserum against GABA showed a large number of labeled somata in the antennal lobe; these somata were located exclusively in the lateral cell cluster. Stained neurites innervating all antennal-lobe glomeruli, including the male-specific macroglomerular complex, suggested a prominent role of GABA in processing olfactory information, including signals from pheromones, interspecifically acting odors, and plant odors. Fibers in two antennocerebral tracts (the middle and dorsal antennocerebral tract) exhibited prominent GABA immunoreactivity. Double-labeling experiments revealed that immunostaining for three neuropeptides, viz., A-type allatostatin, Manduca sexta allatotropin, and FMRFamide-related peptides, was largely colocalized with GABA in cell bodies of the lateral cell cluster. The general absence of peptide immunostaining in the antennocerebral tracts strongly indicated that these peptides were colocalized with GABA in local interneurons of the antennal lobe. In contrast, tachykinin-related peptides occurred in a distinct population of local antennal-lobe neurons that did not exhibit GABA immunostaining. Thus, local interneurons that were not GABAergic were present in the moth antennal lobe.

Clonal analysis ofDrosophilaantennal lobe neurons: diverse neuronal architectures in the lateral neuroblast lineage

The antennal lobe (AL) is the primary structure in the Drosophila brain that relays odor information from the antennae to higher brain centers. The characterization of uniglomerular projection neurons (PNs) and some local interneurons has facilitated our understanding of olfaction; however, many other AL neurons remain unidentified. Because neuron types are mostly specified by lineage and temporal origins, we use the MARCM techniques with a set of enhancer-trap GAL4 lines to perform systematical lineage analysis to characterize neuron morphologies, lineage origin and birth timing in the three AL neuron lineages that contain GAL4-GH146-positive PNs: anterodorsal, lateral and ventral lineages. The results show that the anterodorsal lineage is composed of pure uniglomerular PNs that project through the inner antennocerebral tract. The ventral lineage produces uniglomerular and multiglomerular PNs that project through the middle antennocerebral tract. The lateral lineage generates multiple types of neurons, including uniglomeurlar PNs, diverse atypical PNs, various types of AL local interneurons and the neurons that make no connection within the ALs. Specific neuron types in all three lineages are produced in specific time windows, although multiple neuron types in the lateral lineage are made simultaneously. These systematic cell lineage analyses have not only filled gaps in the olfactory map, but have also exemplified additional strategies used in the brain to increase neuronal diversity.

Caste- and sex-specific adaptations within the olfactory pathway in the brain of the ant Camponotus floridanus

Olfaction plays a key role in mediating ant behavior, and ant societies are characterized by caste- and sex-specific division of labor. We propose that caste- and sex-specific adaptations in the olfactory pathway promote differences in olfactory behavior. This study compares olfactory centers in the brain of large (major) workers, small (minor) workers, virgin queens, and males of the carpenter ant Camponotus floridanus. The number of glomeruli in the antennal lobe was similar in the female castes, although the glomerular volumes differed. Males had ∼45% fewer glomeruli compared to females (∼258 and ∼434) and one antennal sensory tract was absent. A dual output pathway to the mushroom bodies was present in males. In contrast to females, however, the number of glomeruli connected to the medial antennocerebral tract was substantially smaller than those associated with the lateral tract. All glomeruli in the male antennal lobe contained serotonergic processes, whereas in the female castes glomeruli in the large tract six cluster lacked serotonergic innervations. We conclude that differences in general glomerular organization are subtle among the female castes, but sex-specific differences in the number, connectivity and neuromodulatory innervation of glomeruli are substantial and likely to underlie differences in olfactory processing and learning.

The compartment structures of the antennal lobe in the ant,Aphaenogaster smythiesi japonica

Pheromones are important cues for social insects such as ants. As a first step in elucidation of pheromonal information processing mechanisms in the myrmicine ant, we investigated the morphological structure of the antennal lobe. Using autofluorescence imaging, labeling of neuronal filamentous actin, and reduced silver impregnation staining, the antennal lobe was found to consist of five compartments that, each received input from a different antennal sensory tract. Two major tracts of projection neurons, the medial and lateral antenno-cerebral tract (m- and 1-ACT), originated from a different region of the antennal lobe. The m-ACT originated from the posterior part of the antennal lobe whereas the 1-ACT originated from the anterior part. These results demonstrate a spatial segregation of function within the antennal lobe.

Organization of the olfactory pathway and odor processing in the antennal lobe of the ant Camponotus floridanus

Ants rely heavily on olfaction for communication and orientation. Here we provide the first detailed structure-function analyses within an ant's central olfactory system asking whether in the carpenter ant, Camponotus floridanus, the olfactory pathway exhibits adaptations to processing many pheromonal and general odors. Using fluorescent tracing, confocal microscopy, and 3D-analyses we demonstrate that the antennal lobe (AL) contains up to approximately 460 olfactory glomeruli organized in seven distinct clusters innervated via seven antennal sensory tracts. The AL is divided into two hemispheres regarding innervation of glomeruli by either projection neurons (PNs) with axons leaving via the medial (m) or lateral (l) antennocerebral tract (ACT). M- and l-ACT PNs differ in their target areas in the mushroom-body calyx and lateral horn. Three additional ACTs project to the lateral protocerebrum only. We analyzed odor processing in AL glomeruli by retrograde loading of PNs with Fura-2 dextran and fluorimetric calcium imaging. Odor responses were reproducible and comparable across individuals. Calcium responses to pheromonal and nonpheromonal odors were very sensitive (10(-11) dilution) and patterns were partly overlapping, indicating that processing of both odor classes is not spatially segregated within the AL. Response patterns to the main trail-pheromone component nerolic acid remained stable over a wide range of intensities (7-8 log units), while response durations increased indicating that odor quality is maintained by a stable pattern and intensity is mainly encoded in response durations. The structure-function analyses contribute new insights into important aspects of odor processing in a highly advanced insect olfactory system.

Anatomical organization of antennal lobe projection neurons in the moth Heliothis virescens

A prerequisite for understanding how odor information is coded in the central nervous system is to know the morphology and spatial relationship of the principal neurons forming the olfactory pathways. The present account provides an anatomical description of the morphology of the neuronal connections between the antennal lobe and the protocerebrum in the moth Heliothis virescens, a species used for studies of olfactory processing and learning. Intracellular labeling and antennal lobe focal injections with dextran fluorescent markers were combined with neuropil immunostaining and three-dimensional reconstructions. The experiments revealed four antennocerebral tracts, the inner, middle, outer, and dorsomedial, and eight morphological types of projection neurons in addition to a neuron with an unpaired median soma in the subesophageal ganglion. Multiglomerular projection neurons, present in all but the dorsomedial antennocerebral tract, project in several olfactory foci of the protocerebral neuropil. With few exceptions, these neurons do not innervate the calyces of the mushroom body. Uniglomerular projection neurons appear most numerous in the inner antennocerebral tract but are also present in the outer and dorsomedial tracts. These neurons always ramify in the calyces of the mushroom body and in the lateral horn. The projection areas of the neurons following different tracts are largely separated in the secondary olfactory centers. This is most evident in the lateral horn, whereas, in the calyces, the axonal ramifications are more intermingled. The mushroom body architecture, revealed by neuropil immunolabeling, showed striking similarities to that of other lepidopteran species as well as insects of other taxa.

Dual olfactory pathway in the honeybee, Apis mellifera

The antennal lobes (ALs) are the primary olfactory centers in the insect brain. In the AL of the honeybee, olfactory glomeruli receive input via four antennal sensory tracts (T1-4). Axons of projection neurons (PNs) leave the AL via several antenno-cerebral tracts (ACTs). To assign the input-output connectivity of all glomeruli, we investigated the spatial relationship of the antennal tracts and two prominent AL output tracts (medial and lateral ACT) mainly formed by uniglomerular (u) PNs using fluorescent tracing, confocal microscopy, and 3D analyses. Furthermore, we investigated the projections of all ACTs in higher olfactory centers, the mushroom-bodies (MB) and lateral horn (LH). The results revealed a clear segregation of glomeruli into two AL hemispheres specifically supplied by PNs of the medial and lateral ACT. PNs of the lateral ACT innervate glomeruli in the ventral-rostral AL and primarily receive input from T1 (plus a few glomeruli from T2 and T3). PNs of the medial ACT innervate glomeruli in the dorsal-caudal hemisphere, and mainly receive input from T3 (plus a few glomeruli from T2 and T4). The PNs of the m- and l-ACT terminate in different areas of the MB calyx and LH and remain largely segregated. Tracing of three mediolateral (ml) ACTs mainly formed by multiglomerular PNs revealed terminals in distinct compartments of the LH and in three olfactory foci within the lateral protocerebrum. The results indicate that olfactory input in the honeybee is processed via two separate, mainly uPN pathways to the MB calyx and LH and several pathways to the lateral protocerebrum.

Distribution of neuropeptides in the primary olfactory center of the heliothine moth Heliothis virescens

Neuropeptides are a diverse widespread class of signaling substances in the nervous system. As a basis for the analysis of peptidergic neurotransmission in the insect olfactory system, we have studied the distribution of neuropeptides in the antennal lobe of the moth Heliothis virescens. Immunocytochemical experiments with antisera recognizing A-type allatostatins (AST-As), Manduca sexta allatotropin (Mas-AT), FMRFamide-related peptides (FaRPs), and tachykinin-related peptides (TKRPs) have shown that members of all four peptide families are present in local interneurons of the antennal lobe. Whereas antisera against AST-As, Mas-AT, and FaRPs give similar staining patterns characterized by dense meshworks of processes confined to the core of all antennal-lobe glomeruli, TKRPs are present only in neurons with blebby processes distributed throughout each glomerulus. In addition to local neurons, a pair of centrifugal neurons with cell bodies in the lateral subesophageal ganglion, arborizations in the antennal lobe, and projections in the inner antenno-cerebral tracts exhibits tachykinin immunostaining. Double-label immunofluorescence has detected the co-localization of AST-As, Mas-AT, and FaRPs in certain local interneurons, whereas TKRPs occurs in a distinct population. MALDI-TOF mass spectrometry has revealed nearly 50 mass peaks in the antennal lobe. Seven of these masses (four AST-As, two N-terminally extended FLRFamides, and Mas-AT) match known moth neuropeptides. The data thus show that local interneurons of the moth antennal lobe are highly differentiated with respect to their neuropeptide content. The antennal lobe therefore represents an ideal preparation for the future analysis of peptide signaling in insect brain.

Tracheal development in the Drosophila brain is constrained by glial cells

The Drosophila brain is tracheated by the cerebral trachea, a branch of the first segmental trachea of the embryo. During larval stages the cerebral trachea splits into several main (primary) branches that grow around the neuropile, forming a perineuropilar tracheal plexus (PNP) at the neuropile surface. Five primary tracheal branches whose spatial relationship to brain compartments is relatively invariant can be distinguished, although the exact trajectories and branching pattern of the brain tracheae are surprisingly variable. Immunohistochemical and electron microscopic studies demonstrate that all brain tracheae grow in direct contact with the glial cell processes that surround the neuropile. To investigate the effect of glia on tracheal development, embryos and larvae lacking glial cells as a result of a genetic mutation or a directed ablation were analyzed. In these animals, the tracheal branching pattern was highly abnormal. In particular, the number of secondary branches entering the central neuropile was increased. Wild-type larvae possess only two central tracheae, typically associated with the mushroom body and the antennocerebral tract. In larvae lacking glial cells, six to ten tracheal branches penetrate the neuropile in a variable pattern. This finding indicates that glia-derived signals constrained tracheal growth in the Drosophila brain and restrict the number of branches entering the neuropile.

Spatio‐temporal Ca2+ dynamics of moth olfactory projection neurones

We studied the Ca2+ dynamics of odour-evoked glomerular patterns in the antennal lobe of the moth Spodoptera littoralis using optical imaging. Here we selectively stained a large population of antennal lobe output neurones, projection neurones, by retrograde filling with FURA-dextran from the inner antennocerebral tract in the protocerebrum. Different plant-associated odorants evoked distributed patterns of activated glomeruli that were odour dependent and repeatable. These patterns were, however, dynamic during the period of odour exposure. Temporal responses differed across glomeruli and were stimulus dependent. Next we examined how the correlations between patterns evoked by different odorants changed with time. Initially, responses to structurally similar compounds were highly correlated, whereas responses to structurally different compounds differed. Within the period of odour exposure (1 s) we found a significant reduction in similarity of responses evoked by different odours, irrespective of initial similarity, whereas trial-to-trial correlations remained high. Our results suggested an ability for coarse classification at the initial encounter with an odour source. With time, however, the discrimination ability increases and structurally similar odours can be distinguished.