Dynamics of odor-source localization: Insights from real-time odor plume recordings and head-motion tracking in freely moving mice.

Animals navigating turbulent odor plumes exhibit a rich variety of behaviors, and employ efficient strategies to locate odor sources. A growing body of literature has started to probe this complex task of localizing airborne odor sources in walking mammals to further our understanding of neural encoding and decoding of naturalistic sensory stimuli. However, correlating the intermittent olfactory information with behavior has remained a long-standing challenge due to the stochastic nature of the odor stimulus. We recently reported a method to record real-time olfactory information available to freely moving mice during odor-guided navigation, hence overcoming that challenge. Here we combine our odor-recording method with head-motion tracking to establish correlations between plume encounters and head movements. We show that mice exhibit robust head-pitch motions in the 5-14Hz range during an odor-guided navigation task, and that these head motions are modulated by plume encounters. Furthermore, mice reduce their angles with respect to the source upon plume contact. Head motions may thus be an important part of the sensorimotor behavioral repertoire during naturalistic odor-source localization.

The spiking output of the mouse olfactory bulb encodes large-scale temporal features of natural odor environments.

In natural odor environments, odor travels in plumes. Odor concentration dynamics change in characteristic ways across the width and length of a plume. Thus, spatiotemporal dynamics of plumes have informative features for animals navigating to an odor source. Population activity in the olfactory bulb (OB) has been shown to follow odor concentration across plumes to a moderate degree (Lewis et al., 2021). However, it is unknown whether the ability to follow plume dynamics is driven by individual cells or whether it emerges at the population level. Previous research has explored the responses of individual OB cells to isolated features of plumes, but it is difficult to adequately sample the full feature space of plumes as it is still undetermined which features navigating mice employ during olfactory guided search. Here we released odor from an upwind odor source and simultaneously recorded both odor concentration dynamics and cellular response dynamics in awake, head-fixed mice. We found that longer timescale features of odor concentration dynamics were encoded at both the cellular and population level. At the cellular level, responses were elicited at the beginning of the plume for each trial, signaling plume onset. Plumes with high odor concentration elicited responses at the end of the plume, signaling plume offset. Although cellular level tracking of plume dynamics was observed to be weak, we found that at the population level, OB activity distinguished whiffs and blanks (accurately detected odor presence versus absence) throughout the duration of a plume. Even ~20 OB cells were enough to accurately discern odor presence throughout a plume. Our findings indicate that the full range of odor concentration dynamics and high frequency fluctuations are not encoded by OB spiking activity. Instead, relatively lower-frequency temporal features of plumes, such as plume onset, plume offset, whiffs, and blanks, are represented in the OB.

Dynamics of odor-source localization: Insights from real-time odor plume recordings and head-motion tracking in freely moving mice.

Animals navigating turbulent odor plumes exhibit a rich variety of behaviors, and employ efficient strategies to locate odor sources. A growing body of literature has started to probe this complex task of localizing airborne odor sources in walking mammals to further our understanding of neural encoding and decoding of naturalistic sensory stimuli. However, correlating the intermittent olfactory information with behavior has remained a long-standing challenge due to the stochastic nature of the odor stimulus. We recently reported a method to record real-time olfactory information available to freely moving mice during odor-guided navigation, hence overcoming that challenge. Here we combine our odor-recording method with head-motion tracking to establish correlations between plume encounters and head movements. We show that mice exhibit robust head-pitch motions in the 5-14Hz range during an odor-guided navigation task, and that these head motions are modulated by plume encounters. Furthermore, mice orient towards the odor source upon plume contact. Head motions may thus be an important part of the sensorimotor behavioral repertoire during naturalistic odor-source localization.

Mechanisms of octopus arm search behavior without visual feedback.

The octopus coordinates multiple, highly flexible arms with the support of a complex distributed nervous system. The octopus's suckers, staggered along each arm, are employed in a wide range of behaviors. Many of these behaviors, such as foraging in visually occluded spaces, are executed under conditions of limited or absent visual feedback. In coordinating unseen limbs with seemingly infinite degrees of freedom across a variety of adaptive behaviors, the octopus appears to have solved a significant control problem facing the field of soft-bodied robotics. To study the strategies that the octopus uses to find and capture prey within unseen spaces, we designed and 3D printed visually occluded foraging tasks and tracked arm motion as the octopus attempted to find and retrieve a food reward. By varying the location of the food reward within these tasks, we can characterize how the arms and suckers adapt to their environment to find and capture prey. We compared these results to simulated experimental conditions performed by a model octopus arm to isolate the primary mechanisms driving our experimental observations. We found that the octopus relies on a contact-based search strategy that emerges from local sucker coordination to simplify the control of its soft, highly flexible limbs.

Lessons for Robotics From the Control Architecture of the Octopus.

Biological and artificial agents are faced with many of the same computational and mechanical problems, thus strategies evolved in the biological realm can serve as inspiration for robotic development. The octopus in particular represents an attractive model for biologically-inspired robotic design, as has been recognized for the emerging field of soft robotics. Conventional global planning-based approaches to controlling the large number of degrees of freedom in an octopus arm would be computationally intractable. Instead, the octopus appears to exploit a distributed control architecture that enables effective and computationally efficient arm control. Here we will describe the neuroanatomical organization of the octopus peripheral nervous system and discuss how this distributed neural network is specialized for effectively mediating decisions made by the central brain and the continuous actuation of limbs possessing an extremely large number of degrees of freedom. We propose top-down and bottom-up control strategies that we hypothesize the octopus employs in the control of its soft body. We suggest that these strategies can serve as useful elements in the design and development of soft-bodied robotics.

A Machine Learning Approach for Detecting Vicarious Trial and Error Behaviors.

Vicarious trial and error behaviors (VTEs) indicate periods of indecision during decision-making, and have been proposed as a behavioral marker of deliberation. In order to understand the neural underpinnings of these putative bridges between behavior and neural dynamics, researchers need the ability to readily distinguish VTEs from non-VTEs. Here we utilize a small set of trajectory-based features and standard machine learning classifiers to identify VTEs from non-VTEs for rats performing a spatial delayed alternation task (SDA) on an elevated plus maze. We also show that previously reported features of the hippocampal field potential oscillation can be used in the same types of classifiers to separate VTEs from non-VTEs with above chance performance. However, we caution that the modest classifier success using hippocampal population dynamics does not identify many trials where VTEs occur, and show that combining oscillation-based features with trajectory-based features does not improve classifier performance compared to trajectory-based features alone. Overall, we propose a standard set of features useful for trajectory-based VTE classification in binary decision tasks, and support previous suggestions that VTEs are supported by a network including, but likely extending beyond, the hippocampus.

Plume Dynamics Structure the Spatiotemporal Activity of Mitral/Tufted Cell Networks in the Mouse Olfactory Bulb.

Although mice locate resources using turbulent airborne odor plumes, the stochasticity and intermittency of fluctuating plumes create challenges for interpreting odor cues in natural environments. Population activity within the olfactory bulb (OB) is thought to process this complex spatial and temporal information, but how plume dynamics impact odor representation in this early stage of the mouse olfactory system is unknown. Limitations in odor detection technology have made it difficult to measure plume fluctuations while simultaneously recording from the mouse's brain. Thus, previous studies have measured OB activity following controlled odor pulses of varying profiles or frequencies, but this approach only captures a subset of features found within olfactory plumes. Adequately sampling this feature space is difficult given a lack of knowledge regarding which features the brain extracts during exposure to natural olfactory scenes. Here we measured OB responses to naturally fluctuating odor plumes using a miniature, adapted odor sensor combined with wide-field GCaMP6f signaling from the dendrites of mitral and tufted (MT) cells imaged in olfactory glomeruli of head-fixed mice. We precisely tracked plume dynamics and imaged glomerular responses to this fluctuating input, while varying flow conditions across a range of ethologically-relevant values. We found that a consistent portion of MT activity in glomeruli follows odor concentration dynamics, and the strongest responding glomeruli are the best at following fluctuations within odor plumes. Further, the reliability and average response magnitude of glomerular populations of MT cells are affected by the flow condition in which the animal samples the plume, with the fidelity of plume following by MT cells increasing in conditions of higher flow velocity where odor dynamics result in intermittent whiffs of stronger concentration. Thus, the flow environment in which an animal encounters an odor has a large-scale impact on the temporal representation of an odor plume in the OB. Additionally, across flow conditions odor dynamics are a major driver of activity in many glomerular networks. Taken together, these data demonstrate that plume dynamics structure olfactory representations in the first stage of odor processing in the mouse olfactory system.

Using Head-Mounted Ethanol Sensors to Monitor Olfactory Information and Determine Behavioral Changes Associated with Ethanol-Plume Contact during Mouse Odor-Guided Navigation.

Olfaction guides navigation and decision-making in organisms from multiple animal phyla. Understanding how animals use olfactory cues to guide navigation is a complicated problem for two main reasons. First, the sensory cues used to guide animals to the source of an odor consist of volatile molecules, which form plumes. These plumes are governed by turbulent air currents, resulting in an intermittent and spatiotemporally varying olfactory signal. A second problem is that the technologies for chemical quantification are cumbersome and cannot be used to detect what the freely moving animal senses in real time. Understanding how the olfactory system guides this behavior requires knowing the sensory cues and the accompanying brain signals during navigation. Here, we present a method for real-time monitoring of olfactory information using low-cost, lightweight sensors that robustly detect common solvent molecules, like alcohols, and can be easily mounted on the heads of freely behaving mice engaged in odor-guided navigation. To establish the accuracy and temporal response properties of these sensors we compared their responses with those of a photoionization detector (PID) to precisely controlled ethanol stimuli. Ethanol-sensor recordings, deconvolved using a difference-of-exponentials kernel, showed robust correlations with the PID signal at behaviorally relevant time, frequency, and spatial scales. Additionally, calcium imaging of odor responses from the olfactory bulbs (OBs) of awake, head-fixed mice showed strong correlations with ethanol plume contacts detected by these sensors. Finally, freely behaving mice engaged in odor-guided navigation showed robust behavioral changes such as speed reduction that corresponded to ethanol plume contacts.

A selective role for the mPFC during choice and deliberation, but not spatial memory retention over short delays.

Important interactions between memory and decision-making processes are required to maintain high-levels of spatial working memory task performance. Past research reveals that the medial prefrontal cortex (mPFC) and hippocampus (HPC) are both vital structures involved in these processes. Recent evidence suggests that interactions between these two structures are dynamic and task dependent. However, there exists uncertainty surrounding the specific conditions that recruit mPFC contributions to these tasks, specifically regarding its role in retaining information online during delay periods. To address this issue, we tested rats on a spatial-delayed alternation task in which we utilized a closed-loop optogenetic system to transiently disrupt mPFC activity during different task epochs (delay, choice, return). By analyzing the effects of mPFC disruption on choice accuracy and a deliberative behavior known as vicarious-trial-and-error (VTE), our study revealed several interesting findings regarding the role of the mPFC in spatial-working memory tasks. The main findings include: (a) choice accuracy in the spatial-delayed alternation (SDA) task was impaired when the mPFC was disrupted during the choice epoch and not delay or return epochs, (b) mPFC disruption resulted in a non-epoch specific reduction in VTE occurrence which correlated with impairments in task performance. Taken together, findings from this study suggest that, during spatial decision-making, contributions made by the mPFC are specific to points of deliberation and choice (not delay), and that VTEs are a deliberative behavior which relies on intact mPFC function.

Many Paths to the Same Goal: Balancing Exploration and Exploitation during Probabilistic Route Planning.

During self-guided behaviors, animals identify constraints of the problems they face and adaptively employ appropriate strategies (Marsh, 2002). In the case of foraging, animals must balance sensory-guided exploration of an environment with memory-guided exploitation of known resource locations. Here, we show that animals adaptively shift cognitive resources between sensory and memory systems during foraging to optimize route planning under uncertainty. We demonstrate this using a new, laboratory-based discovery method to define the strategies used to solve a difficult route optimization scenario, the probabilistic "traveling salesman" problem (Raman and Gill, 2017; Fuentes et al., 2018; Mukherjee et al., 2019). Using this system, we precisely manipulated the strength of prior information as well as the complexity of the problem. We find that rats are capable of efficiently solving this route-planning problem, even under conditions with unreliable prior information and a large space of possible solutions. Through analysis of animals' trajectories, we show that they shift the balance between exploiting known locations and searching for new locations of rewards based on the predictability of reward locations. When compared with a Bayesian search, we found that animal performance is consistent with an approach that adaptively allocates cognitive resources between sensory processing and memory, enhancing sensory acuity and reducing memory load under conditions in which prior information is unreliable. Our findings establish new approaches to understand neural substrates of natural behavior as well as the rational development of biologically inspired approaches for complex real-world optimization.

Taken out of Context: A Novel Cognitive Role for a Premotor Circuit.

In this issue of Neuron, Wu et al. (2020) provide evidence of a novel role for the premotor cortex in maintaining the context-dependent information necessary for mice to solve a delayed match to sample task.

Balancing Extrasynaptic Excitation and Synaptic Inhibition within Olfactory Bulb Glomeruli.

Glutamatergic transmission in the brain typically occurs at well-defined synaptic connections, but increasing evidence indicates that neural excitation can also occur through activation of "extrasynaptic" glutamate receptors. Here, we investigated the underlying mechanisms and functional properties of extrasynaptic signals that are part of a feedforward path of information flow in the olfactory bulb. This pathway involves glutamatergic interneurons, external tufted cells (eTCs), that are excited by olfactory sensory neurons (OSNs) and in turn excite output mitral cells (MCs) extrasynaptically. Using pair-cell and triple-cell recordings in rat bulb slices (of either sex), combined with ultrastructural approaches, we first present evidence that eTC-to-MC signaling results from "spillover" of glutamate released at eTC synapses onto GABAergic periglomerular (PG) cells in glomeruli. Thus, feedforward excitation is an indirect result of and must cooccur with activation of inhibitory circuitry. Next, to examine the dynamics of the competing signals, we assayed the relationship between the number of spikes in eTCs and excitation of MCs or PG cells in pair-cell recordings. This showed that extrasynaptic excitation in MCs is very weak due to single spikes but rises sharply and supralinearly with increasing spikes, differing from sublinear behavior for synaptic excitation of PG cells. Similar dynamics leading to a preference for extrasynaptic excitation were also observed during recordings of extrasynaptic and inhibitory currents in response to OSN input of increasing magnitude. The observed alterations in the balance between extrasynaptic excitation and inhibition in glomeruli with stimulus strength could underlie an intraglomerular mechanism for olfactory contrast enhancement.

Algorithms for Olfactory Search across Species.

Localizing the sources of stimuli is essential. Most organisms cannot eat, mate, or escape without knowing where the relevant stimuli originate. For many, if not most, animals, olfaction plays an essential role in search. While microorganismal chemotaxis is relatively well understood, in larger animals the algorithms and mechanisms of olfactory search remain mysterious. In this symposium, we will present recent advances in our understanding of olfactory search in flies and rodents. Despite their different sizes and behaviors, both species must solve similar problems, including meeting the challenges of turbulent airflow, sampling the environment to optimize olfactory information, and incorporating odor information into broader navigational systems.

Mice Develop Efficient Strategies for Foraging and Navigation Using Complex Natural Stimuli.

The ability to shift between multiple decision-making strategies during natural behavior allows animals to strike a balance between flexibility and efficiency. We investigated odor-guided navigation by mice to understand how decision-making strategies are balanced during a complex natural behavior. Mice navigated to odor sources in an open arena using naturally fluctuating airborne odor cues as their positions were recorded precisely in real time. When mice had limited prior experience of source locations, their search behavior was consistent with a gradient ascent algorithm that utilized directional cues in the plume to navigate to the odor source. Gradient climbing was effective because the arena size allowed animals to conduct their search mainly within the odor plume, with frequent odor contacts. With increased experience, mice shifted their strategy from this flexible, sensory-driven search behavior to a more efficient and stereotyped foraging approach that varied little in response to odor plumes. This study demonstrates that mice use prior knowledge to adaptively balance flexibility and efficiency during complex behavior guided by dynamic natural stimuli.

ϒ spike-field coherence in a population of olfactory bulb neurons differentiates between odors irrespective of associated outcome.

Studies in different sensory systems indicate that short spike patterns within a spike train that carry items of sensory information can be extracted from the overall train by using field potential oscillations as a reference (Kayser et al., 2012; Panzeri et al., 2014). Here we test the hypothesis that the local field potential (LFP) provides the temporal reference frame needed to differentiate between odors regardless of associated outcome. Experiments were performed in the olfactory system of the mouse (Mus musculus) where the mitral/tufted (M/T) cell spike rate develops differential responses to rewarded and unrewarded odors as the animal learns to associate one of the odors with a reward in a go-no go behavioral task. We found that coherence of spiking in M/T cells with the ϒ LFP (65 to 95 Hz) differentiates between odors regardless of the associated behavioral outcome of odor presentation.

Precise detection of direct glomerular input duration by the olfactory bulb.

Sensory neuron input to the olfactory bulb (OB) was activated precisely for different durations with blue light in mice expressing channelrhodopsin-2 in olfactory sensory neurons. Behaviorally the mice discriminated differences of 10 ms in duration of direct glomerular activation. In addition, a subset of mitral/tufted cells in the OB of awake mice responded tonically therefore conveying information on stimulus duration. Our study provides evidence that duration of the input to glomeruli not synchronized to sniffing is detected. This potent cue may be used to obtain information on puffs in odor plumes.

Information for decision-making and stimulus identification is multiplexed in sensory cortex.

In recordings from anterior piriform cortex in awake behaving mice, we found that neuronal firing early in the olfactory pathway simultaneously conveyed fundamentally different information: odor value (is the odor rewarded?) and identity (what is the smell?). Thus, this sensory system performs early multiplexing of information reflecting stimulus-specific characteristics with that used for decision-making.

Temporal processing in the olfactory system: can we see a smell?

Sensory processing circuits in the visual and olfactory systems receive input from complex, rapidly changing environments. Although patterns of light and plumes of odor create different distributions of activity in the retina and olfactory bulb, both structures use what appears on the surface similar temporal coding strategies to convey information to higher areas in the brain. We compare temporal coding in the early stages of the olfactory and visual systems, highlighting recent progress in understanding the role of time in olfactory coding during active sensing by behaving animals. We also examine studies that address the divergent circuit mechanisms that generate temporal codes in the two systems, and find that they provide physiological information directly related to functional questions raised by neuroanatomical studies of Ramon y Cajal over a century ago. Consideration of differences in neural activity in sensory systems contributes to generating new approaches to understand signal processing.

Functional properties of cortical feedback projections to the olfactory bulb.

Sensory perception is not a simple feed-forward process, and higher brain areas can actively modulate information processing in "lower" areas. We used optogenetic methods to examine how cortical feedback projections affect circuits in the first olfactory processing stage, the olfactory bulb. Selective activation of back projections from the anterior olfactory nucleus/cortex (AON) revealed functional glutamatergic synaptic connections on several types of bulbar interneurons. Unexpectedly, AON axons also directly depolarized mitral cells (MCs), enough to elicit spikes reliably in a time window of a few milliseconds. MCs received strong disynaptic inhibition, a third of which arises in the glomerular layer. Activating feedback axons in vivo suppressed spontaneous as well as odor-evoked activity of MCs, sometimes preceded by a temporally precise increase in firing probability. Our study indicates that cortical feedback can shape the activity of bulbar output neurons by enabling precisely timed spikes and enforcing broad inhibition to suppress background activity.

Mitral cells in the olfactory bulb are mainly excited through a multistep signaling path.

Within the olfactory system, information flow from the periphery onto output mitral cells (MCs) of the olfactory bulb (OB) has been thought to be mediated by direct synaptic inputs from olfactory sensory neurons (OSNs). Here, we performed patch-clamp measurements in rat and mouse OB slices to investigate mechanisms of OSN signaling onto MCs, including the assumption of a direct path, using electrical and optogenetic stimulation methods that selectively activated OSNs. We found that MCs are in fact not typically activated by direct OSN inputs and instead require a multistep, diffuse mechanism involving another glutamatergic cell type, the tufted cells. The preference for a multistep mechanism reflects the fact that signals arising from direct OSN inputs are drastically shunted by connexin 36-mediated gap junctions on MCs, but not tufted cells. An OB circuit with tufted cells intermediate between OSNs and MCs suggests that considerable processing of olfactory information occurs before its reaching MCs.