Second-order Conditioning Research Articles

Parabrachial Calca neurons mediate second-order conditioning

Learning to associate cues, both directly and indirectly, with biologically significant events is essential for survival. Second-order conditioning (SOC) involves forming an association between a previously reinforced conditioned stimulus (CS1) and a new conditioned stimulus (CS2) without the presence of an unconditioned stimulus (US). The neural substrates mediating SOC, however, remain unclear. Parabrachial Calca neurons, which react to the noxious US, also respond to a CS after pairing with a US, suggesting that Calca neurons mediate SOC. We established an aversive SOC behavioral paradigm in mice and monitored Calca neuron activity via single-cell calcium imaging during conditioning and subsequent recall phases. These neurons were activated by both CS1 and CS2 after SOC. Chemogenetically inhibiting Calca neurons during CS1-CS2 pairing attenuated SOC. Thus, reactivation of the US pathway by a learned CS plays an important role in forming the association between the old and a new CS, promoting the formation of second-order memories.

Chronic social defeat stress gives rise to social avoidance through fear learning

Chronic social defeat stress (CSDS), a widely used rodent model of stress, reliably leads to decreased social interaction in stress susceptible animals. Here, we investigate a role for fear learning in this response using male 129 Sv/Ev mice, a strain that is more vulnerable to CSDS than the commonly used C57BL/6 strain. We first demonstrate that defeated 129 Sv/Ev mice avoid a CD-1 mouse, but not a conspecific, indicating that motivation to socialize is intact in this strain. CD-1 avoidance is characterized by approach behavior that results in running in the opposite direction, activity that is consistent with a threat response. We next test whether CD-1 avoidance is subject to the same behavioral changes found in traditional models of Pavlovian fear conditioning. We find that associative learning occurs across 10 days CSDS, with defeated mice learning to associate the color of the CD-1 coat with threat. This leads to the gradual acquisition of avoidance behavior, a conditioned response that can be extinguished with 7 days of repeated social interaction testing (5 tests/day). Pairing a CD-1 with a tone leads to second-order conditioning, resulting in avoidance of an enclosure without a social target. Finally, we show that social interaction with a conspecific is a highly variable response in defeated mice that may reflect individual differences in generalization of fear to other social targets. Our data indicate that fear conditioning to a social target is a key component of CSDS, implicating the involvement of fear circuits in social avoidance.

New high sensitivity approach for a quantitative visualization of the continuous whole-field stress evolution in digital photoelasticity

Digital photoelasticity is a widely used technique for experimental stress analysis on mechanical models subjected to loading conditions, due to its ability to reveal the stress information through color fringe patterns and consequently demodulate it computationally to achieve a quantitative evaluation. In this process, some limitations persist, such as, expertise dependence, experimental repeatability, sensitivity to load increments, and weak performance under experimental conditions, especially in time-dependent scenarios. In response to these weaknesses, the method “Quantitative Visualization of the Continuous Whole-Field Stress Evolution” introduced the ability to perform a complete demodulation of the fringe patterns generated in time-dependent applications, achieving high performance compared to conventional photoelasticity methods. This paper proposes a new highly sensitive approach for a quantitative visualization of the continuous whole-field stress evolution in digital photoelasticity by incorporating an inverse signal process and a multi-stage strategy. The algorithm aims to ensure the performance under high fringe order conditions while exhibiting high sensitivity in cases where the maximum stress obtained is less than one fringe order. The results presented validate its effectiveness in demodulating full-field stress distributions, especially in scenarios where the loading condition induces fringe orders less than one, thus contributing to the broader applicability of digital photoelasticity.

Chronic Social Defeat Stress Gives Rise to Social Avoidance Through Fear Learning.

Chronic social defeat stress (CSDS), a widely used rodent model of stress, reliably leads to decreased social interaction in stress susceptible animals. Here, we investigate a role for fear learning in this response using 129Sv/Ev mice, a strain that is more vulnerable to CSDS than the commonly used C57BL/6 strain. We first demonstrate that defeated 129Sv/Ev mice avoid a CD-1 mouse, but not a conspecific, indicating that motivation to socialize is intact in this strain. CD-1 avoidance is characterized by approach behavior that results in running in the opposite direction, activity that is consistent with a threat response. We next test whether CD-1 avoidance is subject to the same behavioral changes found in traditional models of Pavlovian fear conditioning. We find that associative learning occurs across 10 days CSDS, with defeated mice learning to associate the color of the CD-1 coat with threat. This leads to the gradual acquisition of avoidance behavior, a conditioned response that can be extinguished with 7 days of repeated social interaction testing (5 tests/day). Pairing a CD-1 with a tone leads to second-order conditioning, resulting in avoidance of an enclosure without a social target. Finally, we show that social interaction with a conspecific is a highly variable response in defeated mice that may reflect individual differences in generalization of fear to other social targets. Our data indicate that fear conditioning to a social target is a key component of CSDS, implicating the involvement of fear circuits in social avoidance.

Acute ethanol disrupts conditioned inhibition in the male rat.

Alcohol can disrupt conditioned sexual inhibition (CSI) established by first-order conditioning in male rats. CSI can also be induced using second-order conditioning, during which male rats are trained to associate a neutral odor with a nonreceptive female. As a result, when given access to two receptive females (one scented and one unscented) during a copulatory preference test, they display CSI toward the scented female. The present study examined the effect of low-to-moderate doses of alcohol on CSI and brain activation following exposure to alcohol and the olfactory cue alone. Sexually-naïve Long-Evans rats received alternate conditioning sessions with unscented receptive or scented (almond extract) non-receptive females. Following the conditioning phase, males were injected with saline, alcohol 0.5g/kg or 1g/kg, 45min before a copulatory test with two receptive females, with one bearing the olfactory cue. Fos activation was later assessed, following exposure to alcohol and the olfactory cue alone, in several brain regions involved in the expression and regulation of male sexual behavior. While males in the saline group displayed sexual avoidance towards the scented female, those injected with alcohol before the copulatory test, regardless of the dose, copulated indiscriminately with both females. Subsequent exposure to alcohol and the olfactory cue alone induced different Fos expression between groups in several brain regions. Low to moderate doses of alcohol disrupt conditioned sexual inhibition in male rats and induce a differential pattern of neural activation, particularly in regions involved in the expression and regulation of sexual behavior.

Cognitive mapping and episodic memory emerge from simple associative learning rules

Episodic memory enables animals to map contexts and environmental features in space and time but is underused in artificial intelligence (AI). Here we show how simple associative learning rules can be expanded to basic episodic memory in AI. We augment an agent-based foraging simulation, ASIMOV, modeled on the simple neuronal circuitry of an invertebrate forager, by adding a novel computational module for simple episodic memory, the Feature Association Matrix (FAM). The FAM is a set of computationally light, graph learning algorithms which functionally resemble the auto- and hetero-associative circuits of the hippocampus for episodic memory. In simulations, FAM enables highly efficient foraging and navigation and shows how higher-order conditioning mechanisms give rise to spatial cognitive mapping by chaining pair-wise associations and encoding them with additional contexts. Thus, FAM demonstrates a biologically inspired, bottom-up enhancement of AI for higher-order cognition.

Cognitive limits of larval Drosophila: testing for conditioned inhibition, sensory preconditioning, and second-order conditioning.

Drosophila larvae are an established model system for studying the mechanisms of innate and simple forms of learned behavior. They have about 10 times fewer neurons than adult flies, and it was the low total number of their neurons that allowed for an electron microscopic reconstruction of their brain at synaptic resolution. Regarding the mushroom body, a central brain structure for many forms of associative learning in insects, it turned out that more than half of the classes of synaptic connection had previously escaped attention. Understanding the function of these circuit motifs, subsequently confirmed in adult flies, is an important current research topic. In this context, we test larval Drosophila for their cognitive abilities in three tasks that are characteristically more complex than those previously studied. Our data provide evidence for (i) conditioned inhibition, as has previously been reported for adult flies and honeybees. Unlike what is described for adult flies and honeybees, however, our data do not provide evidence for (ii) sensory preconditioning or (iii) second-order conditioning in Drosophila larvae. We discuss the methodological features of our experiments as well as four specific aspects of the organization of the larval brain that may explain why these two forms of learning are observed in adult flies and honeybees, but not in larval Drosophila.

Diverse memory paradigms in Drosophila reveal diverse neural mechanisms.

In this review, we aggregated the different types of learning and memory paradigms developed in adult Drosophila and attempted to assess the similarities and differences in the neural mechanisms supporting diverse types of memory. The simplest association memory assays are conditioning paradigms (olfactory, visual, and gustatory). A great deal of work has been done on these memories, revealing hundreds of genes and neural circuits supporting this memory. Variations of conditioning assays (reversal learning, trace conditioning, latent inhibition, and extinction) also reveal interesting memory mechanisms, whereas mechanisms supporting spatial memory (thermal maze, orientation memory, and heat box) and the conditioned suppression of innate behaviors (phototaxis, negative geotaxis, anemotaxis, and locomotion) remain largely unexplored. In recent years, there has been an increased interest in multisensory and multicomponent memories (context-dependent and cross-modal memory) and higher-order memory (sensory preconditioning and second-order conditioning). Some of this work has revealed how the intricate mushroom body (MB) neural circuitry can support more complex memories. Finally, the most complex memories are arguably those involving social memory: courtship conditioning and social learning (mate-copying and egg-laying behaviors). Currently, very little is known about the mechanisms supporting social memories. Overall, the MBs are important for association memories of multiple sensory modalities and multisensory integration, whereas the central complex is important for place, orientation, and navigation memories. Interestingly, several different types of memory appear to use similar or variants of the olfactory conditioning neural circuitry, which are repurposed in different ways.

Roles of feedback and feed-forward networks of dopamine subsystems: insights from Drosophila studies.

Across animal species, dopamine-operated memory systems comprise anatomically segregated, functionally diverse subsystems. Although individual subsystems could operate independently to support distinct types of memory, the logical interplay between subsystems is expected to enable more complex memory processing by allowing existing memory to influence future learning. Recent comprehensive ultrastructural analysis of the Drosophila mushroom body revealed intricate networks interconnecting the dopamine subsystems-the mushroom body compartments. Here, we review the functions of some of these connections that are beginning to be understood. Memory consolidation is mediated by two different forms of network: A recurrent feedback loop within a compartment maintains sustained dopamine activity required for consolidation, whereas feed-forward connections across compartments allow short-term memory formation in one compartment to open the gate for long-term memory formation in another compartment. Extinction and reversal of aversive memory rely on a similar feed-forward circuit motif that signals omission of punishment as a reward, which triggers plasticity that counteracts the original aversive memory trace. Finally, indirect feed-forward connections from a long-term memory compartment to short-term memory compartments mediate higher-order conditioning. Collectively, these emerging studies indicate that feedback control and hierarchical connectivity allow the dopamine subsystems to work cooperatively to support diverse and complex forms of learning.

On the role of higher-order condition of strain gradient plasticity in the cyclic torsion of thin metallic wires: Experiments and modeling

Cyclic torsion tests are performed on micron-scale copper wires with and without surface passivation to study the role of the higher-order condition in the plastic behavior of thin wires under non-proportional loading. A typical strengthening size effect is observed in the symmetric cycles. More obvious strength enhancement exists in the torsional response of passivated copper wires. An unusual Bauschinger effect is found during the loading-unloading cycles, which is more pronounced in passivated wires. The finite element implementation based on Gudmundson's strain gradient plasticity theory is developed for wire torsion to characterize the observed size-dependent phenomena. The higher-order boundary conditions are introduced to simulate the passivated surface. The predicted radial distributions of plastic strain, stress components, and geometrically necessary dislocation density for the passivated and unpassivated wires are given and compared. This work provides a reasonable basis for understanding the role of higher-order conditions of strain gradient plasticity.

Minimal circuit motifs for second-order conditioning in the insect mushroom body.

In well-established first-order conditioning experiments, the concurrence of a sensory cue with reinforcement forms an association, allowing the cue to predict future reinforcement. In the insect mushroom body, a brain region central to learning and memory, such associations are encoded in the synapses between its intrinsic and output neurons. This process is mediated by the activity of dopaminergic neurons that encode reinforcement signals. In second-order conditioning, a new sensory cue is paired with an already established one that presumably activates dopaminergic neurons due to its predictive power of the reinforcement. We explored minimal circuit motifs in the mushroom body for their ability to support second-order conditioning using mechanistic models. We found that dopaminergic neurons can either be activated directly by the mushroom body's intrinsic neurons or via feedback from the output neurons via several pathways. We demonstrated that the circuit motifs differ in their computational efficiency and robustness. Beyond previous research, we suggest an additional motif that relies on feedforward input of the mushroom body intrinsic neurons to dopaminergic neurons as a promising candidate for experimental evaluation. It differentiates well between trained and novel stimuli, demonstrating robust performance across a range of model parameters.

Lipopolysaccharide (LPS) attenuates the primary conditioning of lithium chloride (LiCl)-induced context aversion but not the secondary conditioning of context aversion or taste avoidance

A first-order association can be formed between toxin-induced nausea and a context, as well as nausea and a taste cue. However, comparatively little is understood about second-order associations. The present study examined if the bacterial endotoxin, LPS, could impair the first- and second-order conditioning of context aversion (anticipatory nausea paradigm) and subsequent conditioned taste avoidance (two-bottle task). Adult male Long Evans rats were treated with LiCl (127 mg/kg, intraperitoneal [i.p.]) or vehicle control (NaCl) and then exposed to a distinct context for 4 first-order conditioning trials. LPS (200 μg/kg, i.p.) or NaCl were administered 24 h after each trial. Seventy-two h after the final first-order conditioning trial, rats underwent 2 second-order conditioning trials where they were treated with 2% saccharin (i.p.) and then exposed to the same context. Twenty-four h after the final second-order conditioning trial, rats were tested in a two-bottle task (2 trials), where they were given a choice between water and a palatable 0.2% saccharin solution. LiCl-treated rats demonstrated a context aversion by the 3rd conditioning trial in the anticipatory nausea paradigm. Rats previously exposed to LiCl also displayed a conditioned taste avoidance of saccharin within the two-bottle task. LPS attenuated first-order context aversion but did not alter either second-order context aversion or conditioned taste avoidance in the two-bottle task. This study demonstrated that a secondary association formed within an aversive context could result in a conditioned taste avoidance. Further, LPS may be able to attenuate primary conditioning, but not secondary conditioning.

The Basolateral Amygdala: The Core of a Network for Threat Conditioning, Extinction, and Second-Order Threat Conditioning.

Threat conditioning, extinction, and second-order threat conditioning studied in animal models provide insight into the brain-based mechanisms of fear- and anxiety-related disorders and their treatment. Much attention has been paid to the role of the basolateral amygdala (BLA) in such processes, an overview of which is presented in this review. More recent evidence suggests that the BLA serves as the core of a greater network of structures in these forms of learning, including associative and sensory cortices. The BLA is importantly regulated by hippocampal and prefrontal inputs, as well as by the catecholaminergic neuromodulators, norepinephrine and dopamine, that may provide important prediction-error or learning signals for these forms of learning. The sensory cortices may be required for the long-term storage of threat memories. As such, future research may further investigate the potential of the sensory cortices for the long-term storage of extinction and second-order conditioning memories.

Arbeitsschutz „im Huckepack“ – ein alternatives Konzept zur Ansprache von Unternehmen auf Sicherheits- und Gesundheitsthemen. Teil II: Erprobung in einer Maßnahme für Kleinunternehmen

Occupational safety “piggybacked” – an alternative concept for addressing companies with safety and health issues. Part II: Implementation in a measure for small businesses Background: Entrepreneurs in small businesses represent a challenging but also underestimated target group for occupational safety and health (OSH) prevention activities. Objective: Referring to the article published in ASU 6/2023 on the theoretical foundation of the “piggyback method”, an alternative concept for addressing small business owners on OSH issues is being developed and implemented. Method: OSH issues were linked by content to topics preferred by the target group and presented according to the principles of second-order conditioning. The interactive events were marketed by two chambers and evaluated by an online survey with a random sample of n = 40. Results: The procedure and implementation were rated positively overall by the participants. The OSH issues (target stimulus) were rated as just as relevant as the lack of skilled workers and self-care (context stimuli). Conclusions: The results speak for the usefulness of the “piggyback method” for OSH prevention activities. However, the results are not sufficient as a proof of effectiveness; this requires further practical tests and evaluations. Keywords: occupational safety and health – small businesses – entrepreneurs – evaluation

A Review of Backward Higher-Order Conditioning: Implications for a Pavlovian Conditioning Analysis of Stimulus Equivalence.

Stimulus equivalence (SE) is demonstrated when participants exposed to conditional discrimination training pass tests for reflexivity, symmetry, transitivity, and equivalence (symmetry combined with transitivity). Most theorists attribute the origin of SE to operant processes, but some argue that it results from Pavlovian conditioning. Symmetry is problematic for the latter hypothesis because it seems to require excitatory backward conditioning. However, equivalence tests resemble backward sensory preconditioning (BSP) and backward second-order conditioning (BSOC), two well-established processes. A review of associationistic theories of BSP and BSOC showed that the temporal coding hypothesis (TCH) explains outcomes that other associationistic theories cannot explain (i.e., BSOC and BSP effects after first-order conditioning with delay vs. trace conditioning and forward vs. backward conditioning). The TCH assumes that organisms encode the temporal attributes of stimulus events (e.g., order and interval duration) and this temporal information is integrated across separate phases of training. The TCH seems compatible with a behavioral analysis if direct stimulus control replaces the notion of temporal maps. The TCH perspective does not seem applicable to SE because SE tests are not predictive tasks. This suggests that SE is fundamentally different from BSP and BSOC and a Pavlovian conditioning analysis of SE is inadequate. This conclusion is consistent with previous criticism of a Pavlovian account of SE according to which Pavlovian conditioning cannot be interpreted as stimulus substitution.

The standard reconsolidation protocol for auditory fear-conditioning does not account for fear to the test context.

Research on memory reconsolidation has relied heavily on the use of Pavlovian auditory cued-fear conditioning. Here, an auditory cue (CS) is paired with a footshock (US) and the CS is later able to evoke a freezing response when presented alone. Some treatments, when administered to conditioned subjects immediately following a CS-alone (memory reactivation) trial, can attenuate the freezing they display on subsequent CS-alone (test) trials, in the absence of the treatment. This reduction in conditioned freezing is usually taken as evidence that the treatment disrupts post-reactivation reconsolidation of the memory trace representing the pairing of CS and US. We suggest an alternative interpretation that may account, either in whole or in part, for the attenuated freezing. The standard reconsolidation protocol (SRP) for auditory fear-conditioning has a design feature that results in second-order conditioning of fear to the test context, as this context is paired with the fear-evoking CS on the reactivation trial. Since freezing during the CS on the test will reflect the compound influence of contextual-fear and cued-fear, a post-reactivation treatment might attenuate freezing on the test by disrupting consolidation of second-order contextual-fear conditioning, even if it has little or no effect on the stability of the original cued-fear memory. This experiment confirmed that rats tested according to the SRP, in which the reactivation and test trials occur in the same context, freeze more on the test trial than rats that receive the reactivation and test trials in different contexts. This confound could lead to false-positive evidence of disrupted reconsolidation if it is not avoided or minimized, which can be accomplished with a modified protocol.

Octopamine neurons mediate reward signals in social learning in an insect

Social learning is found in many animals, but its mechanisms are not understood. We previously showed that a cricket that was trained to observe a conspecific staying at a drinking apparatus exhibited an increased preference for the odor of that drinking apparatus. Here we investigated a hypothesis that this learning is achieved by second-order conditioning (SOC), i.e., by associating conspecifics at a drinking bottle with water reward during group drinking in the rearing stage and then associating an odor with a conspecific in training. Injection of an octopamine receptor antagonist before training or testing impaired the learning or response to the learned odor, as we reported for SOC, thereby supporting the hypothesis. Notably, the SOC hypothesis predicts that octopamine neurons that respond to water in the group-rearing stage also respond to a conspecific in training, without the learner itself drinking water, and such mirror-like activities mediate social learning. This awaits future investigation.

Hierarchical architecture of dopaminergic circuits enables second-order conditioning in Drosophila.

Dopaminergic neurons with distinct projection patterns and physiological properties compose memory subsystems in a brain. However, it is poorly understood whether or how they interact during complex learning. Here, we identify a feedforward circuit formed between dopamine subsystems and show that it is essential for second-order conditioning, an ethologically important form of higher-order associative learning. The Drosophila mushroom body comprises a series of dopaminergic compartments, each of which exhibits distinct memory dynamics. We find that a slow and stable memory compartment can serve as an effective 'teacher' by instructing other faster and transient memory compartments via a single key interneuron, which we identify by connectome analysis and neurotransmitter prediction. This excitatory interneuron acquires enhanced response to reward-predicting odor after first-order conditioning and, upon activation, evokes dopamine release in the 'student' compartments. These hierarchical connections between dopamine subsystems explain distinct properties of first- and second-order memory long known by behavioral psychologists.

Ideal quadratic fermionic point state with multiple band degeneracy

As the study of topological states witnessed rapid progress and fast development, the current research has been expanded from conventional linear dispersion into high order conditions. In comparison with the linear type, high order topological elements feature multiple exotic properties, such as large topological charge, peculiar Berry phase, and chiral surface states, which could lead to the finding of other new physics. In this study, we present a realistic material candidate hosting an ideal quadratic fermionic point state with multiple band degeneracy. Based on first principles calculation and effective model argument, the origin of the quadratic order is analyzed, and its dispersion conditions are examined. Due to the clean band structure, prominent surface states are observed and they exhibit both large energy variation and broad spatial distribution. These beneficial features are very advantageous for future experimental investigations. This work can significantly enhance the research on high order topological states and, in particular, the proposed material can provide an effective platform for quadratic fermionic point states.

New second-order optimality conditions in sub-riemannian geometry

We study the geometry of the second-order expansion of the extended end-point map for the sub-Riemannian geodesic problem. Translating the geometric reality into equations we derive new second-order necessary optimality conditions in sub-Riemannian Geometry. In particular, we find an ODE for velocity of an abnormal sub-Riemannian geodesics. It allows to divide abnormal minimizers into two classes, which we propose to call 2-normal and 2-abnormal extremals. In the 2-normal case the above ODE completely determines the velocity of a curve, while in the 2-abnormal case the velocity is undetermined at some, or at all points. With some enhancement of the presented results it should be possible to prove the regularity of all 2-normal extremals (the 2-abnormal case seems to require study of higher-order conditions) thus making a step towards solving the problem of smoothness of sub-Riemannian abnormal geodesics. As a by-product we present a new derivation of Goh conditions. We also prove that the assumptions weaker than these used in [Boarotto, Monti, Palmurella, 2020] to derive third-order Goh conditions, imply piece-wise-C2 regularity of an abnormal extremal. Marty me i Zuzi