Yuriy Zhurov

From Hunger to Satiety: Reconfiguration of a Feeding Network by Aplysia Neuropeptide Y

A shift in motivational state often produces behavioral change, but the underlying mechanisms are poorly understood. In the marine mollusc, Aplysia californica, feeding-induced transition from a hunger to satiation state leads to a slowdown and an eventual termination of feeding. Because the multifunctional feeding network generates both ingestion and the competing response, egestion, it is possible that the transition from a hunger to a satiety state is associated with network reconfiguration that results in production of fewer ingestive and more egestive responses. Chronic electrophysiological recordings in free-feeding Aplysia showed that as the meal progressed, food elicited fewer ingestive responses and simultaneously increased the number of egestive responses. Injections of Aplysia neuropeptide Y (apNPY) reduced food intake and slowed down the rate of ingestion. apNPY was localized to buccal-ganglion afferents originating in the gut-innervating esophageal nerve (EN), a nerve involved both in satiation and in the generation of egestive programs. During EN stimulation, apNPY was released in the feeding circuit. Importantly, stimulation of the cerebral-buccal interneuron-2, a command-like interneuron that is activated by food and normally elicits ingestive responses, elicited egestive responses in the presence of apNPY. This was accompanied by increased activity of the egestion-promoting interneuron B20 and decreased activity in the ingestion-promoting interneuron B40. Thus, apNPYergic reconfiguration of the feeding central pattern generator plays a role in the gradual transition from hunger to satiety states. More generally, changes in the motivational states may involve not only simple network inhibition but may also require network reconfiguration.

Changes of Internal State Are Expressed in Coherent Shifts of Neuromuscular Activity in Aplysia Feeding Behavior

The multitasking central pattern generator (CPG) that drives consummatory feeding behaviors of Aplysia can produce ingestive, egestive, and intermediate motor programs. External stimuli trigger the programs but, remarkably, do not directly specify which type of program is produced. Rather, recent work has proposed, the type of program is determined by the internal network state of the CPG that has developed in response to the previous history of the stimulation. Here we have tested a key prediction of this network-state hypothesis. If the network state has a real existence and governs real functional behavior, changes in the state should be seen as coherent, coordinated changes along many dimensions of interneuron and motor neuron activity, muscle contraction, and ultimately movement, that underlie functional behavior. In reduced neuromuscular preparations, we elicited repetitive motor programs by continued stimulation of the esophageal nerve while recording the firing of motor neurons B8, B15, B16, B4/5, and B48, and contractions of the accessory radula closer and I7-I10 muscles that respectively close and open the animals food-grasping organ, the radula. Using sonomicrometric techniques, we similarly recorded the movement of the radula in the complete buccal mass. Successive esophageal nerve programs indeed exhibited clear progressive changes in motor neuron firing, muscle contractions, and the phasing of radula movements within each cycle, from an initially intermediate or even ingestive character to a strongly egestive character. We conclude that the Aplysia feeding CPG really has a coherent internal network state whose dynamics are likely to be reflected in the real behavior of the animal.

Variability of Motor Neuron Spike Timing Maintains and Shapes Contractions of the Accessory Radula Closer Muscle of Aplysia

The accessory radula closer (ARC) muscle of Aplysia has long been studied as a typical “slow” muscle, one that would be assumed to respond only to the overall, integrated spike rate of its motor neurons, B15 and B16. The precise timing of the individual spikes should not much matter. However, but real B15 and B16 spike patterns recorded in vivo show great variability that extends down to the timing of individual spikes. By replaying these real as well as artificially constructed spike patterns into ARC muscles in vitro, we examined the consequences of this spike-level variability for contraction. Replaying the same pattern several times reproduces precisely the same contraction shape: the B15/B16–ARC neuromuscular transform is deterministic. However, varying the timing of the spikes produces very different contraction shapes and amplitudes. The transform in fact operates at an interface between “fast” and “slow” regimens. It is fast enough that the timing of individual spikes greatly influences the detailed contraction shape. At the same time, slow integration of the spike pattern through the nonlinear transform allows the variable spike timing to determine also the overall contraction amplitude. Indeed, the variability appears to be necessary to maintain the contraction amplitude at a robust level. This phenomenon is tuned by neuromodulators that tune the speed and nonlinearity of the transform. Thus, the variable timing of individual spikes does matter, in at least two, functionally significant ways, in this “slow” neuromuscular system.

Identification of a new neuropeptide precursor reveals a novel source of extrinsic modulation in the feeding system of aplysia

The Aplysia feeding system is advantageous for investigating the role of neuropeptides in behavioral plasticity. One family of Aplysia neuropeptides is the myomodulins (MMs), originally purified from one of the feeding muscles, the accessory radula closer (ARC). However, two MMs, MMc and MMe, are not encoded on the only known MM gene. Here, we identify MM gene 2 (MMG2), which encodes MMc and MMe and four new neuropeptides. We use matrix-assisted laser desorption/ionization time-of-flight mass spectrometry to verify that these novel MMG2-derived peptides (MMG2-DPs), as well as MMc and MMe, are synthesized from the precursor. Using antibodies against the MMG2-DPs, we demonstrate that neuronal processes that stain for MMG2-DPs are found in the buccal ganglion, which contains the feeding network, and in the buccal musculature including the ARC muscle. Surprisingly, however, no immunostaining is observed in buccal neurons including the ARC motoneurons. In situ hybridization reveals only few MMG2-expressing neurons that are mostly located in the pedal ganglion. Using immunohistochemical and electrophysiological techniques, we demonstrate that some of these pedal neurons project to the buccal ganglion and are the likely source of the MMG2-DP innervation of the feeding network and musculature. We show that the MMG2-DPs are bioactive both centrally and peripherally: they bias egestive feeding programs toward ingestive ones, and they modulate ARC muscle contractions. The multiple actions of the MMG2-DPs suggest that these peptides play a broad role in behavioral plasticity and that the pedal-buccal projection neurons that express them are a novel source of extrinsic modulation of the feeding system of Aplysia.

Motor outputs in a multitasking network: relative contributions of inputs and experience-dependent network states.

Network outputs elicited by a specific stimulus may differ radically depending on the momentary network state. One class of networks states-experience-dependent states-is known to operate in numerous networks, yet the fundamental question concerning the relative role that inputs and states play in determining the network outputs remains to be investigated in a behaviorally relevant manner. Because previous work indicated that in the isolated nervous system the motor outputs of the Aplysia feeding network are affected by experience-dependent states, we sought to establish the behavioral relevance of these outputs. We analyzed the phasing of firing of radula opening motoneurons (B44 and B48) relative to other previously characterized motoneurons. We found that the overall pattern of motoneuronal firing corresponds to the phasing of movements during feeding behavior, thus indicating a behavioral relevance of network outputs. Previous studies suggested that network inputs act to trigger a response rather than to shape its characteristics, with the latter function being fulfilled by network states. We show this is an oversimplification. In a rested state, different inputs elicited distinct responses, indicating that inputs not only trigger but also shape the responses. However, depending on the combination of inputs and states, responses were either dramatically altered by the network state or were indistinguishable from those observed in the rested state. We suggest that the relative contributions of inputs and states are dynamically regulated and, rather than being fixed, depend on the specifics of states and inputs.

Predicting Adaptive Behavior in the Environment from Central Nervous System Dynamics

To generate adaptive behavior, the nervous system is coupled to the environment. The coupling constrains the dynamical properties that the nervous system and the environment must have relative to each other if adaptive behavior is to be produced. In previous computational studies, such constraints have been used to evolve controllers or artificial agents to perform a behavioral task in a given environment. Often, however, we already know the controller, the real nervous system, and its dynamics. Here we propose that the constraints can also be used to solve the inverse problem—to predict from the dynamics of the nervous system the environment to which they are adapted, and so reconstruct the production of the adaptive behavior by the entire coupled system. We illustrate how this can be done in the feeding system of the sea slug Aplysia. At the core of this system is a central pattern generator (CPG) that, with dynamics on both fast and slow time scales, integrates incoming sensory stimuli to produce ingestive and egestive motor programs. We run models embodying these CPG dynamics—in effect, autonomous Aplysia agents—in various feeding environments and analyze the performance of the entire system in a realistic feeding task. We find that the dynamics of the system are tuned for optimal performance in a narrow range of environments that correspond well to those that Aplysia encounter in the wild. In these environments, the slow CPG dynamics implement efficient ingestion of edible seaweed strips with minimal sensory information about them. The fast dynamics then implement a switch to a different behavioral mode in which the system ignores the sensory information completely and follows an internal “goal,” emergent from the dynamics, to egest again a strip that proves to be inedible. Key predictions of this reconstruction are confirmed in real feeding animals.