ABSTRACT
Natural behaviors are a coordinated symphony of motor acts that drive reafferent (self-induced) sensory activation. Individual sensors cannot disambiguate exafferent (externally induced) from reafferent sources. Nevertheless, animals readily differentiate between these sources of sensory signals to carry out adaptive behaviors through corollary discharge circuits (CDCs), which provide predictive motor signals from motor pathways to sensory processing and other motor pathways. Yet, how CDCs comprehensively integrate into the nervous system remains unexplored. Here, we use connectomics, neuroanatomical, physiological, and behavioral approaches to resolve the network architecture of two pairs of ascending histaminergic neurons (AHNs) in Drosophila, which function as a predictive CDC in other insects. Both AHN pairs receive input primarily from a partially overlapping population of descending neurons, especially from DNg02, which controls wing motor output. Using Ca2+ imaging and behavioral recordings, we show that AHN activation is correlated to flight behavior and precedes wing motion. Optogenetic activation of DNg02 is sufficient to activate AHNs, indicating that AHNs are activated by descending commands in advance of behavior and not as a consequence of sensory input. Downstream, each AHN pair targets predominantly non-overlapping networks, including those that process visual, auditory, and mechanosensory information, as well as networks controlling wing, haltere, and leg sensorimotor control. These results support the conclusion that the AHNs provide a predictive motor signal about wing motor state to mostly non-overlapping sensory and motor networks. Future work will determine how AHN signaling is driven by other descending neurons and interpreted by AHN downstream targets to maintain adaptive sensorimotor performance.
Subject(s)
Drosophila , Neurons , Animals , Drosophila/physiology , Efferent Pathways , InsectaABSTRACT
Natural behaviors are a coordinated symphony of motor acts which drive self-induced or reafferent sensory activation. Single sensors only signal presence and magnitude of a sensory cue; they cannot disambiguate exafferent (externally-induced) from reafferent sources. Nevertheless, animals readily differentiate between these sources of sensory signals to make appropriate decisions and initiate adaptive behavioral outcomes. This is mediated by predictive motor signaling mechanisms, which emanate from motor control pathways to sensory processing pathways, but how predictive motor signaling circuits function at the cellular and synaptic level is poorly understood. We use a variety of techniques, including connectomics from both male and female electron microscopy volumes, transcriptomics, neuroanatomical, physiological and behavioral approaches to resolve the network architecture of two pairs of ascending histaminergic neurons (AHNs), which putatively provide predictive motor signals to several sensory and motor neuropil. Both AHN pairs receive input primarily from an overlapping population of descending neurons, many of which drive wing motor output. The two AHN pairs target almost exclusively non-overlapping downstream neural networks including those that process visual, auditory and mechanosensory information as well as networks coordinating wing, haltere, and leg motor output. These results support the conclusion that the AHN pairs multi-task, integrating a large amount of common input, then tile their output in the brain, providing predictive motor signals to non-overlapping sensory networks affecting motor control both directly and indirectly.
ABSTRACT
Insects exhibit remarkable sensory and motor capabilities to successfully navigate their environment. As insects move, they activate sensory afferents. Hence, insects are inextricably part of their sensory ecology. Insects must correctly attribute self- versus external sources of sensory activation to make adaptive behavioral choices. This is achieved via corollary discharge circuits (CDCs), motor-to-sensory neuronal pathways providing predictive motor signals to sensory networks to coordinate sensory processing within the context of ongoing behavior. While CDCs provide predictive motor signals, their underlying mechanisms of action and functional consequences are diverse. Here, we describe inferred CDCs and identified corollary discharge interneurons (CDIs) in insects, highlighting their anatomical commonalities and our limited understanding of their synaptic integration into the nervous system. By using connectomics information, we demonstrate that the complexity with which identified CDIs integrate into the central nervous system (CNS) can be revealed.
Subject(s)
Sensation , Sensory Receptor Cells , Animals , Sensation/physiology , Central Nervous System/physiology , InsectaABSTRACT
Nervous systems must distinguish sensory signals derived from an animal's own movements (reafference) from environmentally derived sources (exafference). To accomplish this, motor networks producing reafference transmit motor information, via a corollary discharge circuit (CDC), to affected sensory networks, modulating sensory function during behavior. While CDCs have been described in most sensory modalities, none have been observed projecting to an olfactory pathway. In moths, two mesothoracic to deutocerebral histaminergic neurons (MDHns) project from flight sensorimotor centers in the mesothoracic neuromere to the antennal lobe (AL), where they provide the sole source of histamine (HA), but whether they represent a CDC is unknown. We demonstrate that MDHn spiking activity is positively correlated with wing-motor output and increased before bouts of motor activity, suggesting that MDHns communicate global locomotor state, rather than providing a precisely timed motor copy. Within the AL, HA application sharpened entrainment of projection neuron responses to odor stimuli embedded within simulated wing-beat-induced flows, whereas MDHn axotomy or AL HA receptor (HA-r) blockade reduced entrainment. This finding is consistent with higher-order CDCs, as the MDHns enhanced rather than filtered entrainment of AL projection neurons. Finally, HA-r blockade increased odor detection and discrimination thresholds in behavior assays. These results establish MDHns as a CDC that modulates AL temporal resolution, enhancing odor-guided behavior. MDHns thus appear to represent a higher-order CDC to an insect olfactory pathway; this CDC's unique nature highlights the importance of motor-to-sensory signaling as a context-specific mechanism that fine-tunes sensory function.
Subject(s)
Flight, Animal , Histamine/pharmacology , Olfactory Pathways/physiology , Olfactory Receptor Neurons/physiology , Wings, Animal/physiology , Animals , Manduca , Olfactory Bulb/cytology , Olfactory Bulb/drug effects , Olfactory Bulb/physiology , Olfactory Pathways/drug effects , Olfactory Receptor Neurons/cytology , Olfactory Receptor Neurons/drug effects , Wings, Animal/drug effectsABSTRACT
Nervous systems must adapt to shifts in behavioural ecology. One form of adaptation is neural exaptation, in which neural circuits are co-opted to perform additional novel functions. Here, we describe the co-option of a motor-to-somatosensory circuit into an olfactory network. Many moths beat their wings during odour-tracking, whether walking or flying, causing strong oscillations of airflow around the antennae, altering odour plume structure. This self-induced sensory stimulation could impose selective pressures that influence neural circuit evolution, specifically fostering the emergence of corollary discharge circuits. In Manduca sexta, a pair of mesothoracic to deutocerebral histaminergic neurons (MDHns), project from the mesothoracic neuromere to both antennal lobes (ALs), the first olfactory neuropil. Consistent with a hypothetical role in providing the olfactory system with a corollary discharge, we demonstrate that the MDHns innervate the ALs of advanced and basal moths, but not butterflies, which differ in wing beat and flight pattern. The MDHns probably arose in crustaceans and in many arthropods innervate mechanosensory areas, but not the olfactory system. The MDHns, therefore, represent an example of architectural exaptation, in which neurons that provide motor output information to mechanosensory regions have been co-opted to provide information to the olfactory system in moths.
Subject(s)
Flight, Animal , Manduca/physiology , Neurons/physiology , Olfactory Bulb/physiology , Animals , Histamine , Smell/physiology , Wings, AnimalABSTRACT
Neural circuits projecting information from motor to sensory pathways are common across sensory domains. These circuits typically modify sensory function as a result of motor pattern activation; this is particularly so in cases where the resultant behavior affects the sensory experience or its processing. However, such circuits have not been observed projecting to an olfactory pathway in any species despite well characterized active sampling behaviors that produce reafferent mechanical stimuli, such as sniffing in mammals and wing beating in the moth Manduca sexta. In this study we characterize a circuit that connects a flight sensory-motor center to an olfactory center in Manduca. This circuit consists of a single pair of histamine immunoreactive (HA-ir) neurons that project from the mesothoracic ganglion to innervate a subset of ventral antennal lobe (AL) glomeruli. Furthermore, within the AL we show that the M. sexta histamine B receptor (MsHisClB) is exclusively expressed by a subset of GABAergic and peptidergic LNs, which broadly project to all olfactory glomeruli. Finally, the HA-ir cell pair is present in fifth stage instar larvae; however, the absence of MsHisClB-ir in the larval antennal center indicates that the circuit is incomplete prior to metamorphosis and importantly prior to the expression of flight behavior. Although the functional consequences of this circuit remain unknown, these results provide the first detailed description of a circuit that interconnects an olfactory system with motor centers driving flight behaviors including odor-guided flight.
Subject(s)
Manduca/anatomy & histology , Olfactory Pathways/physiology , Sensory Receptor Cells/physiology , Smell/physiology , Animals , Drosophila Proteins/metabolism , FMRFamide/metabolism , Histamine/metabolism , Insect Hormones/metabolism , Metamorphosis, Biological , Microscopy, Confocal , Neuropeptides/metabolism , Olfactory Bulb/cytology , Olfactory Pathways/injuries , Receptors, Histamine/metabolism , gamma-Aminobutyric Acid/metabolismABSTRACT
As odor concentration increases, primary olfactory network representations expand in spatial distribution, temporal complexity and duration. However, the direct relationship between concentration dependent odor representations and the psychophysical thresholds of detection and discrimination is poorly understood. This relationship is absolutely critical as thresholds signify transition points whereby representations become meaningful to the organism. Here, we matched stimulus protocols for psychophysical assays and intracellular recordings of antennal lobe (AL) projection neurons (PNs) in the moth Manduca sexta to directly compare psychophysical thresholds and the output representations they elicit. We first behaviorally identified odor detection and discrimination thresholds across an odor dilution series for a panel of structurally similar odors. We then characterized spatiotemporal spiking patterns across a population of individually filled and identified AL PNs in response to those odors at concentrations below, at, and above identified thresholds. Using spatial and spatiotemporal based analyses we observed that each stimulus produced unique representations, even at sub-threshold concentrations. Mean response latency did not decrease and the percent glomerular activation did not increase with concentration until undiluted odor. Furthermore, correlations between spatial patterns for odor decreased, but only significantly with undiluted odor. Using time-integrated Euclidean distance (ED) measures, we determined that added spatiotemporal information was present at the discrimination but not detection threshold. This added information was evidenced by an increase in integrated distance between the sub-detection and discrimination threshold concentrations (of the same odor) that was not present in comparison of the sub-detection and detection threshold. After consideration of delays for information to reach the AL we find that it takes ~120-140 ms for the AL to output identity information. Overall, these results demonstrate that as odor concentration increases, added information about odor identity is embedded in the spatiotemporal representation at the discrimination threshold.
ABSTRACT
Animals use behaviors to actively sample the environment across a broad spectrum of sensory domains. These behaviors discretize the sensory experience into unique spatiotemporal moments, minimize sensory adaptation, and enhance perception. In olfaction, behaviors such as sniffing, antennal flicking, and wing beating all act to periodically expose olfactory epithelium. In mammals, it is thought that sniffing enhances neural representations; however, the effects of insect wing beating on representations remain unknown. To determine how well the antennal lobe (AL) produces odor dependent representations when wing beating effects are simulated, we used extracellular methods to record neural units and local field potentials (LFPs) from moth AL. We recorded responses to odors presented as prolonged continuous stimuli or periodically as 20 and 25 Hz pulse trains designed to simulate the oscillating effects of wing beating around the antennae during odor guided flight. Using spectral analyses, we show that ~25% of all recorded units were able to entrain to "pulsed stimuli"; this includes pulsed blanks, which elicited the strongest overall entrainment. The strength of entrainment to pulse train stimuli was dependent on molecular features of the odorants, odor concentration, and pulse train duration. Moreover, units showing pulse tracking responses were highly phase locked to LFPs during odor stimulation, indicating that unit-LFP phase relationships are stimulus-driven. Finally, a Euclidean distance-based population vector analysis established that AL odor representations are more robust, peak more quickly, and do not show adaptation when odors were presented at the natural wing beat frequency as opposed to prolonged continuous stimulation. These results suggest a general strategy for optimizing olfactory representations, which exploits the natural rhythmicity of wing beating by integrating mechanosensory and olfactory cues at the level of the AL.
ABSTRACT
Sensory systems sample the external world actively, within the context of self-motion induced disturbances. Mammals sample olfactory cues within the context of respiratory cycles and have adapted to process olfactory information within the time frame of a single sniff cycle. In plume tracking insects, it remains unknown whether olfactory processing is adapted to wing beating, which causes similar physical effects as sniffing. To explore this we first characterized the physical properties of our odor delivery system using hotwire anemometry and photo ionization detection, which confirmed that odor stimuli were temporally structured. Electroantennograms confirmed that pulse trains were tracked physiologically. Next, we quantified odor detection in moths in a series of psychophysical experiments to determine whether pulsing odor affected acuity. Moths were first conditioned to respond to a target odorant using Pavlovian olfactory conditioning. At 24 and 48 h after conditioning, moths were tested with a dilution series of the conditioned odor. On separate days odor was presented either continuously or as 20 Hz pulse trains to simulate wing beating effects. We varied pulse train duty cycle, olfactometer outflow velocity, pulsing method, and odor. Results of these studies, established that detection was enhanced when odors were pulsed. Higher velocity and briefer pulses also enhanced detection. Post hoc analysis indicated enhanced detection was the result of a significantly lower behavioral response to blank stimuli when presented as pulse trains. Since blank responses are a measure of false positive responses, this suggests that the olfactory system makes fewer errors (i.e. is more reliable) when odors are experienced as pulse trains. We therefore postulate that the olfactory system of Manduca sexta may have evolved mechanisms to enhance odor detection during flight, where the effects of wing beating represent the norm. This system may even exploit temporal structure in a manner similar to sniffing.
Subject(s)
Manduca/physiology , Odorants , Smell/physiology , Wings, Animal/physiology , Animals , Conditioning, Classical , PsychophysicsABSTRACT
Nitric oxide (NO) is thought to play an important neuromodulatory role in olfaction. We are using the hawkmoth Manduca sexta to investigate the function of NO signaling in the antennal lobe (AL; the primary olfactory network in invertebrates). We have found previously that NO is present at baseline levels, dramatically increases in response to odor stimulation, and alters the electrophysiology of AL neurons. It is unclear, however, how these effects contribute to common features of olfactory systems such as olfactory learning and memory, odor detection and odor discrimination. In this study, we used chemical detection and a behavioral approach to further examine the function of NO in the AL. We found that basal levels of NO fluctuate with the daily light cycle, being higher during the nocturnal active period. NO also appears to be necessary for short-term olfactory memory. NO does not appear to affect odor detection, odor discrimination between dissimilar odorants, or learning acquisition. These findings suggest a modulatory role for NO in the timing of olfactory-guided behaviors.
Subject(s)
Manduca/physiology , Nitric Oxide Synthase/metabolism , Nitric Oxide/metabolism , Animals , Association Learning , Brain/metabolism , Circadian Rhythm , Female , Memory, Short-Term , Olfactory PerceptionABSTRACT
The transient oscillatory model of odor identity encoding seeks to explain how odorants with spatially overlapped patterns of input into primary olfactory networks can be discriminated. This model provides several testable predictions about the distributed nature of network oscillations and how they control spike timing. To test these predictions, 16 channel electrode arrays were placed within the antennal lobe (AL) of the moth Manduca sexta. Unitary spiking and multi site local field potential (LFP) recordings were made during spontaneous activity and in response to repeated presentations of an odor panel. We quantified oscillatory frequency, cross correlations between LFP recording sites, and spike-LFP phase relationships. We show that odor-driven AL oscillations in Manduca are frequency modulating (FM) from â¼100 to 30 Hz; this was odorant and stimulus duration dependent. FM oscillatory responses were localized to one or two recording sites suggesting a localized (perhaps glomerular) not distributed source. LFP cross correlations further demonstrated that only a small (r < 0.05) distributed and oscillatory component was present. Cross spectral density analysis demonstrated the frequency of these weakly distributed oscillations was state dependent (spontaneous activity = 25-55 Hz; odor-driven = 55-85 Hz). Surprisingly, vector strength analysis indicated that unitary phase locking of spikes to the LFP was strongest during spontaneous activity and dropped significantly during responses. Application of bicuculline, a GABA(A) receptor antagonist, significantly lowered the frequency content of odor-driven distributed oscillatory activity. Bicuculline significantly reduced spike phase locking generally, but the ubiquitous pattern of increased phase locking during spontaneous activity persisted. Collectively, these results indicate that oscillations perform poorly as a stimulus-mediated spike synchronizing mechanism for Manduca and hence are incongruent with the transient oscillatory model.
ABSTRACT
Subpopulations of Kenyon cells, the intrinsic neurons of the insect mushroom bodies, are typically sequentially generated by dedicated neuroblasts that begin proliferating during embryogenesis. When present, Class III Kenyon cells are thought to be the first born population of neurons by virtue of the location of their cell somata, farthest from the position of the mushroom body neuroblasts. In the adult tobacco hornworm moth Manduca sexta, the axons of Class III Kenyon cells form a separate Y tract and dorsal and ventral lobelet; surprisingly, these distinctive structures are absent from the larval Manduca mushroom bodies. BrdU labeling and immunohistochemical staining reveal that Class III Kenyon cells are in fact born in the mid-larval through adult stages. The peripheral position of their cell bodies is due to their genesis from two previously undescribed protocerebral neuroblasts distinct from the mushroom body neuroblasts that generate the other Kenyon cell types. These findings challenge the notion that all Kenyon cells are produced solely by the mushroom body neuroblasts, and may explain why Class III Kenyon cells are found sporadically across the insects, suggesting that when present, they may arise through de novo recruitment of neuroblasts outside of the mushroom bodies. In addition, lifelong neurogenesis by both the Class III neuroblasts and the mushroom body neuroblasts was observed, raising the possibility that adult neurogenesis may play a role in mushroom body function in Manduca.
Subject(s)
Cell Differentiation , Manduca/cytology , Mushroom Bodies/cytology , Neurons/cytology , Animals , Larva/cytology , Manduca/growth & development , Mushroom Bodies/physiology , Neurogenesis , Neurons/physiology , Neurons/ultrastructureABSTRACT
Each down stroke of an insect's wings accelerates axial airflow over the antennae. Modeling studies suggest that this can greatly enhance penetration of air and air-born odorants through the antennal sensilla thereby periodically increasing odorant-receptor interactions. Do these periodic changes result in entrainment of neural responses in the antenna and antennal lobe (AL)? Does this entrainment affect olfactory acuity? To address these questions, we monitored antennal and AL responses in the moth Manduca sexta while odorants were pulsed at frequencies from 10-72 Hz, encompassing the natural wingbeat frequency. Power spectral density (PSD) analysis was used to identify entrainment of neural activity. Statistical analysis of PSDs indicates that the antennal nerve tracked pulsed odor up to 30 Hz. Furthermore, at least 50% of AL local field potentials (LFPs) and between 7-25% of unitary spiking responses also tracked pulsed odor up to 30 Hz in a frequency-locked manner. Application of bicuculline (200 muM) abolished pulse tracking in both LFP and unitary responses suggesting that GABA(A) receptor activation is necessary for pulse tracking within the AL. Finally, psychophysical measures of odor detection establish that detection thresholds are lowered when odor is pulsed at 20 Hz. These results suggest that AL networks can respond to the oscillatory dynamics of stimuli such as those imposed by the wing beat in a manner analogous to mammalian sniffing.
ABSTRACT
A central problem facing studies of neural encoding in sensory systems is how to accurately quantify the extent of spatial and temporal responses. In this study, we take advantage of the relatively simple and stereotypic neural architecture found in invertebrates. We combine standard electrophysiological techniques, recently developed population analysis techniques, and novel anatomical methods to form an innovative 4-dimensional view of odor output representations in the antennal lobe of the moth Manduca sexta. This novel approach allows quantification of olfactory responses of characterized neurons with spike time resolution. Additionally, arbitrary integration windows can be used for comparisons with other methods such as imaging. By assigning statistical significance to changes in neuronal firing, this method can visualize activity across the entire antennal lobe. The resulting 4-dimensional representation of antennal lobe output complements imaging and multi-unit experiments yet provides a more comprehensive and accurate view of glomerular activation patterns in spike time resolution.
Subject(s)
Brain Mapping/methods , Brain/physiology , Electrophysiology/methods , Image Processing, Computer-Assisted/methods , Neurons/physiology , Staining and Labeling/methods , Animals , Brain/cytology , Brain Mapping/instrumentation , Data Interpretation, Statistical , Electrophysiology/instrumentation , Fluorescent Dyes , Ganglia, Invertebrate/cytology , Ganglia, Invertebrate/physiology , Male , Manduca , Neurons/cytology , Neurophysiology/instrumentation , Neurophysiology/methods , Olfactory Pathways/cytology , Olfactory Pathways/physiology , Signal Processing, Computer-Assisted , Smell/physiology , Species SpecificityABSTRACT
Studies of olfactory function show that disruption of GABA A receptors within the insect antennal lobe (AL) disrupts discrimination of closely related odors, suggesting that local processing within the AL specifically enhances fine odor discrimination. It remains unclear, however, how extensively AL function has been disrupted in these circumstances. Here we psychophysically characterize the effect of GABA A blockade in the AL of the moth Manduca sexta. We used 2 GABA A antagonists and 3 Pavlovian-based behavioral assays of olfactory function. In all cases, we used matched saline-injected controls in a blind study. Using a stimulus generalization assay, we found that GABA A disruption abolished the differential response to related odors, suggesting that local processing mediates fine odor discrimination. We then assessed the effect of GABA A antagonist on discrimination thresholds. Moths were differentially conditioned to respond to one odor (reinforced conditioned stimulus [CS+]) but not a second (unreinforced conditioning stimulus [CS-]) then tested for a significant differential conditioned response between them across a series of increasing concentrations. Here, GABA A blockade disrupted discrimination of both similar and dissimilar odor pairs as indicated by generally increased discrimination thresholds. Finally, using a detection threshold assay, we established that GABA A blockade also increases detection thresholds. Because detection is a prerequisite of discrimination, this later finding suggests that disrupted discrimination may be due to impairment of the ability to detect. We conclude that the loss of ability to detect and subsequently discriminate is attributable to a loss of ability of the AL to provide a clear neural signal from background.
Subject(s)
Olfactory Bulb/physiology , Receptors, GABA-A/physiology , Sensory Thresholds/physiology , Smell/physiology , Animals , Bicuculline/pharmacology , Conditioning, Classical , Electromyography , Feeding Behavior/drug effects , Female , GABA-A Receptor Antagonists , Male , Manduca , Odorants , Olfactory Bulb/drug effects , Picrotoxin/pharmacology , Sensory Thresholds/drug effects , Smell/drug effectsABSTRACT
What is the spatial and temporal nature of odor representations within primary olfactory networks at the threshold of an animal's ability to discriminate? Although this question is of central importance to olfactory neuroscience, it can only be answered in model systems where neural representations can be measured and discrimination thresholds between odors can be characterized. Here, we establish these thresholds for a panel of odors using a Pavlovian paradigm in the moth Manduca sexta. Moths were differentially conditioned to respond to one odor (CS+) but not another (CS-) using undiluted odorants to minimize salience-dependent learning effects. At 24 and 48 h postconditioning, moths were tested for the presence of a conditioned response (CR) with a blank, then the CS+ and CS- (pseudorandomly) across a 5-log step series of increasing concentration. Results identified discrimination thresholds and established that differential CRs to the CS+ and CS- increased with stimulus concentration. Next, 3 separate groups of moths were differentially conditioned at either one-log step below, at, or one log step above the identified discrimination threshold. At 24 and 48 h postconditioning, moths were tested sequentially with a blank, the concentration used for conditioning, and then undiluted odor. Conditioning at one log step below the discrimination threshold established a CR, indicating both stimulus detection and learning, but was insufficient to establish evidence of discrimination. Moths conditioned at the discrimination threshold were able to discriminate but only when stimulated with undiluted odors, indicating learning, but discrimination measures were hampered. When conditioned above the discrimination threshold, moths had no difficulty in discriminating. These results establish methods for psychophysical characterization of discrimination and indicate that differential conditioning at lowered concentrations biases threshold measures.
Subject(s)
Conditioning, Classical/physiology , Discrimination Learning/physiology , Manduca/physiology , Smell/physiology , Animals , Dose-Response Relationship, Drug , Electromyography , Feeding Behavior/physiology , Female , Male , Odorants , Perception/physiology , Psychophysics , Sex FactorsABSTRACT
The precise relationship between olfactory transduction sensitivity and sensitivity at the level of sensory perception is poorly understood. The goal of this work was to correlate neurophysiological measures of sensory transduction and psychophysical measures of salient odor perception using the moth (Manduca sexta). Moths were conditioned to respond to a single monomolecular odor and then tested across a dilution series. Resulting concentration-response functions were dependent on the conditioning odor and its concentration but not on the moth's sex. Comparison of responsiveness to odor versus an odorless blank provided a statistical indicator of detection threshold. Separate control experiments revealed that conditioned responsiveness to odor was a function of test concentration and independent of the conditioning concentration. Next, electroantennogram (EAG) responses to these odorants and concentrations were recorded. EAG responses were highly correlated with the behavioral results but typically identified lower detection thresholds. Furthermore, significant effects of sex, Sex x Odor, and Sex x Odor x Concentration were observed only within the EAG data, suggesting a mismatch across methods. Possible reasons for and implications of this mismatch are discussed.
Subject(s)
Behavior, Animal/physiology , Manduca/physiology , Olfactory Pathways/physiology , Perception/physiology , Signal Detection, Psychological/physiology , Smell/physiology , Analysis of Variance , Animals , Conditioning, Classical/physiology , Dose-Response Relationship, Drug , Electrophysiology , Evoked Potentials/drug effects , Evoked Potentials/physiology , Female , Male , Odorants , Probability , Sensory Thresholds/physiology , Sex FactorsABSTRACT
Recent evidence suggests that odor-driven responses in the insect antennal lobe (AL) can be modified by associative and nonassociative processes, as has been shown in the vertebrate olfactory bulb. However, the specific network changes that occur in response to olfactory learning remain unknown. To characterize changes in AL network activity during learning, we developed an in vivo protocol in Manduca sexta that allows continuous monitoring of neural ensembles and feeding behavior over the course of olfactory conditioning. Here, we show that Pavlovian conditioning produced a net recruitment of responsive neural units across the AL that persisted after conditioning. Recruitment only occurred when odor reliably predicted food. Conversely, when odor did not predict food, a net loss of responsive units occurred. Simultaneous measures of feeding responses indicated that the treatment-specific patterns of neural recruitment were positively correlated with changes in the insect's behavioral response to odor. In addition to recruitment, conditioning also produced consistent and profound shifts in the temporal responses of 16% of recorded units. These results show that odor representations in the AL are dynamic and related to olfactory memory consolidation. We furthermore provide evidence that the basis of the learning-dependent changes in the AL is not simply an increase in activity in the neural network representing an odorant. Rather, learning produces a restructuring of spatial and temporal components of network responses to odor in the AL.
Subject(s)
Discrimination Learning/physiology , Manduca/physiology , Memory/physiology , Smell/physiology , Animals , Conditioning, Psychological/physiology , Electromyography , Odorants , Olfactory Pathways/physiologyABSTRACT
Behavioral studies of olfactory discrimination and stimulus generalization in many species indicate that the molecular features of monomolecular odorants are important for odor discrimination. Here we evaluate how features, such as carbon chain length and functional group, are represented in the first level of synaptic processing. We recorded antennal lobe ensemble responses in the moth Manduca sexta to repeated 100-ms pulses of monomolecular alcohols and ketones. Most units exhibited a significant change in spike rate in response to most odorants that outlasted the duration of the stimulus. Peristimulus data were then sampled over 780 ms for each pulse of all odorants. Factor analysis was used to assess whether there were groups of units with common response patterns. We found that factors identified and represented activity for clusters of units with common temporal response characteristics. These temporally patterned responses typically spanned 780 ms and were often dependent on carbon chain length and functional group. Furthermore, cross-correlation analysis frequently indicated significant coincident spiking even during spontaneous activity. However, this synchrony occurred mainly between units recorded on the same tetrode. In a final analysis, the Euclidean distance between odor responses was calculated for each pair of odorants using factors as dimensions. The distance between responses for any two odorants was maximized by approximately 240 ms. This time course corresponded to the brief sequence of coordinated bursts across the recorded population. The distance during this period was also a function of systematic differences in molecular features. Results of this Euclidian analysis thus directly correlate to previous behavioral studies of stimulus generalization in M. sexta.