Extending in vitro conditioning in Aplysia to analyze operant and classical processes in the same preparation

Operant and classical conditioning are major processes shaping behavioral responses in all animals. Although the understanding of the mechanisms of classical conditioning has expanded significantly, the understanding of the mechanisms of operant conditioning is more limited. Recent developments in Aplysia are helping to narrow the gap in the level of understanding between operant and classical conditioning, and have raised the possibility of studying the neuronal processes underlying the interaction of operant and classical components in a relatively complex learning task. In the present study, we describe a first step toward realizing this goal, by developing a single in vitro preparation in which both operant and classical conditioning can be studied concurrently. The new paradigm reproduced previously published results, even under more conservative and homogenous selection criteria and tonic stimulation regime. Moreover, the observed learning was resistant to delay, shortening, and signaling of reinforcement.     

A cellular analogue of operant conditioning.

Using the hippocampal-slice preparation, we attempted to model operant conditioning in vitro by reinforcing pyramidal cell bursting responses with local micropressure applications of transmitters and drugs. The same injections were administered independently of bursting to provide a "noncontingent" control for direct pharmacological stimulation or facilitation of firing. The results suggested that the bursting responses of individual CA1 pyramidal neurons may be reinforced in a dose-related manner by response-contingent (but not noncontingent) injections of dopamine and the selective dopamine D2 agonist, N-0923. N-0924, a stereoisomer of N-0923 that is largely devoid of D2-agonist activity, failed to reinforce CA1 bursting. Burst-contingent injections of the excitatory neurotransmitter glutamate also failed to reinforce CA1 bursting; indeed, the glutamate applications (whether contingent or random) reduced the likelihood of bursts while increasing the frequency of solitary spikes. Reinforcement delays exceeding 200 ms largely eliminated the reinforcing efficacy of the D2 agonist N-0437 in CA1 operant conditioning. The results are consistent with the suggestion that the behaviorally reinforcing effects of dopaminergic agents can be modeled in vitro in the hippocampal-slice preparation.     

Identification of a reinforcement pathway necessary for operant conditioning of head waving in Aplysia californica

The marine mollusc Aplysia californica exhibits a wide range of nonassociative and associative forms of learning. Recently, we found that the learning repertoire of Aplysia includes operant conditioning (Cook & Carew, 1986, 1989b). The behavior we examined is a naturally occurring, side-to-side head-waving response used by Aplysia in seeking food, obtaining a foothold, and egg laying. Aplysia can be operantly conditioned to reduce head-waving to one side of their body if such a response results in exposure to bright uniform-field illumination, which the animals find aversive. An essential step toward achieving a mechanistic understanding of operant conditioning is to identify and characterize the reinforcement pathway used during the learning. Toward this end, we wished to determine which of the peripheral visual pathways in Aplysia are critical for performance of the operant task. Previous experiments indicated that photic input from the optic and rhinophore nerves functionally inhibited motor neurons that participate in the operant response (head-waving), while photic input from the oral veil nerves excited these same motor neurons (Cook & Carew, 1989c). These findings suggested the hypothesis that one or both of these pathways could play an important role in mediating reinforcement during training. To explore this possibility we operantly trained animals that had received chronic bilateral transections of either the optic and rhinophore nerves or the oral veil nerves C1-C3 (in conjunction with transection of the optic and rhinophore nerves). We found that operant conditioning was not disrupted by ablation of input from the eyes and rhinophores. By contrast, ablation of input from the oral veil (together with that from the eyes and rhinophores) abolished operant conditioning. Thus, the oral veil nerves play a critical modulatory role in operant conditioning of head-waving. This observation further suggested that photic input from the oral veil is conveyed to the CNS via the oral veil nerves. In a final experiment we confirmed that stimulation of the oral veil with light evokes increased afferent activity in the oral veil nerves C1-C2. These results support the idea that the oral veil nerves contain processes that are critical components of the reinforcement pathway for operant conditioning of head-waving.     

Design decisions in a Pascal-based operant conditioning system

FREDL, a Pascal-based system for control of operant experiments, has been implemented on the IBM PC computer. In the foreground, recurring operations, such as decrementing counters that serve as local clocks, are controlled by a dictionary specifying automatic properties of user defined variables, and random logic is implemented by a user-written Pascal overlay. In the background, the remainder of the program controls communication with the user, output of data to disk, and production of cumulative records.     

Reinforcement in an in vitro analog of appetitive classical conditioning of feeding behavior in Aplysia: blockade by a dopamine antagonist

In a recently developed in vitro analog of appetitive classical conditioning of feeding in Aplysia, the unconditioned stimulus (US) was electrical stimulation of the esophageal nerve (En). This nerve is rich in dopamine (DA)-containing processes, which suggests that DA mediates reinforcement during appetitive conditioning. To test this possibility, methylergonovine was used to antagonize DA receptors. Methylergonovine (1 nM) blocked the pairing-specific increase in fictive feeding that is usually induced by in vitro classical conditioning. The present results and previous observation that methylergonovine also blocks the effects of contingent reinforcement in an in vitro analog of appetitive operant conditioning suggest that DA mediates reinforcement for appetitive associative conditioning of feeding in Aplysia.     

Transference in view of a classical conditioning model

This article presents a qualitative metasynthetic study, addressing 33 transference case studies, that investigates the interrelationship of the transference concept from psychoanalysis and cognitive-behavioral concepts in an attempt to construct a theoretical platform for clinical integration. Relationship between categories analysis was used to compare Luborsky's (1998) transference components (wish, response from other, and response of self) and cognitive-behavioral ones. Results showed reciprocal relations between transference and classical conditioning. Furthermore, explicit occurrences of distorted thinking due to overgeneralization were found in more than 90% of the cases. A conceptual model describes transference as a conditioned response activated by thematic conditioning, a particular case of classical conditioning that repeatedly pairs a given interpersonal situation with internal thematic stimuli, thus shaping the person's narrative. Theoretical and practical implications are discussed as well.     

Formulation of a behavior system for sexual conditioning

Empirical and conceptual developments that led to the formulation of a behavior system for the sexual conditioning of male Japanese quail are described. Initial efforts concentrated on conditioning with localized conditioned stimuli and on identifying behavioral indices of conditioning. Later, learning about species-typical cues and about contextual cues was also explored, and it became evident that different types of cues control different aspects of sexual behavior. The results were used to formulate a behavior system containing both response and stimulus dimensions. In this system, contextual cues and local cues are assumed to elicit only general search behavior unconditionally. In contrast, unconditioned responses to species-typical cues of a female quail include general search, focal search, and copulatory behavior. General search, focal search, and copulatory behavior can become conditioned to local cues. Conditioning can also modify focal search behavior elicited by species-typical cues and can result in various modulatory influences between different types of stimuli. The behavior system approach provides a framework for organizing the diverse sexual conditioning effects and suggests future directions for investigation.     

Discriminated response and incentive processes in operant conditioning: a two-factor model of stimulus control

Understanding stimulus control generated in instrumental learning requires the direct investigation of discriminated response and reinforcer (incentive) processes acquired exclusively through the response-reinforcer contingencies operating on complex (multicomponent) baselines. Two series of stimulus-compounding studies accomplished this direct investigation. In one series, the independent variable was the relative reinforcement between schedule components; in the second series, it was relative response rate between components. Stimulus-compounding tests revealed that response and incentive processes enhanced each other when in agreement, counteracted each other when in opposition, and produced intermediate results when only one factor was operating. This pattern of results led to the conclusion that these factors were algebraically combining and to the development of a response/incentive matrix reflecting these dynamics. This two-factor analysis was extended to the peak-shift effect in stimulus generalization experiments and to the generation of inhibitory control. Two decades of stimulus compounding and peak-shift research were organized within this two-factor framework, extending this traditional approach to learning to active research areas heretofore not systematically considered in these terms.     

Multiple memory stores and operant conditioning: a rationale for memory's complexity

Why does the brain contain more than one memory system? Genetic algorithms can play a role in elucidating this question. Here, model animals were constructed containing a dorsal striatal layer that controlled actions, and a ventral striatal layer that controlled a dopaminergic learning signal. Both layers could gain access to three modeled memory stores, but such access was penalized as energy expenditure. Model animals were then selected on their fitness in simulated operant conditioning tasks. Results suggest that having access to multiple memory stores and their representations is important in learning to regulate dopamine release, as well as in contextual discrimination. For simple operant conditioning, as well as stimulus discrimination, hippocampal compound representations turned out to suffice, a counterintuitive result given findings that hippocampal lesions tend not to affect performance in such tasks. We argue that there is in fact evidence to support a role for compound representations and the hippocampus in even the simplest conditioning tasks.     

A microcomputer system for controlling classical conditioning experiments

A microcomputer-based laboratory system for controlling stimulus presentations and data acquisition in classical conditioning experiments is described. The system comprises an Intel 386/486-based microcomputer and a commercially obtained low-cost counter/timer board with input/output lines for stimulus timing and external device control. A simple, yet versatile custom-designed structured programming language is provided for performing an unlimited number of stimulus configurations and their sequences. In electrophysiological studies, the system can be flexibly connected to computer-controlled signal conditioning systems for the amplification and filtering of multiunit and evoked field potential responses and to high-speed data acquisition systems for sampling and analyzing the responses. The costs of reserving an entire microcomputer for experiment control are well compensated for by the simplicity and efficiency of programming and transportability of the control protocols between different setups and laboratories. Furthermore, a data acquisition and analysis system most suitable for the aims of a research project can be selected.     

Recent advances in operant conditioning technology: a versatile and affordable computerized touchscreen system.

We report the construction of a new operant chamber that incorporates modern computer, touchscreen, and display technologies. An LCD display was housed in the front wall of a lightweight Plexiglas chamber. An Apple eMac computer was used to present visual stimuli on the monitor and to control other chamber events. Responses to the stimuli were recorded using a transparent resistive-type touchscreen that overlaid the monitor. The resulting system is simple and inexpensive to construct but powerful and flexible enough to explore a broad range of issues in animal learning and behavior.     

Hippocampal theta-dependent eyeblink classical conditioning: coordination of a distributed learning system

Richard F. Thompson's study of the neurobiological substrates of learning and memory has been a career-long endeavor, chosen early and pursued with uncompromising depth and breadth. His systematic mapping of the major brain systems and mechanisms involved in eyeblink classical conditioning (EBCC) established the essential role of the cerebellum. Investigations of the interactions between the hippocampus and cerebellum are critically important to this literature, given the essential involvement of these structures in trace EBCC as well as an important modulatory role of the hippocampus in delay EBCC. Hippocampal theta (3-7Hz) oscillations are known to reflect a functional state that influences both the timing of unit firing and the potential for neural plasticity in the hippocampus and other structures. Herein we present a brief summary of research demonstrating the behavioral enhancement due to theta and the underlying neurobiological correlates in both hippocampus and cerebellum during EBCC. Hippocampal and cerebellar local field potentials (LFPs) show that these distantly interconnected brain structures become precisely synchronized when conditions favor rapid behavioral acquisition. Our results suggest a major role for theta in coordinating the widely distributed memory system for trace EBCC. These and other important findings reflect Thompson's own work and his early-career mentoring of scientists whose contributions to the EBCC literature have ensured his major and lasting impact on the neurobiology of learning and memory.