Threatening stimuli elicit a sequential cardiac pattern in arthropods.

In order to cope with the challenges of living in dynamic environments, animals rapidly adjust their behaviors in coordination with different physiological responses. Here, we studied whether threatening visual stimuli evoke different heart rate patterns in arthropods and whether these patterns are related with defensive behaviors. We identified two sequential phases of crab's cardiac response that occur with a similar timescale to that of the motor arrest and later escape response. The first phase was modulated by low salience stimuli and persisted throughout spaced stimulus presentation. The second phase was modulated by high-contrast stimuli and reduced by repetitive stimulus presentation. The overall correspondence between cardiac and motor responses suggests that the first cardiac response phase might be related to motor arrest while the second to the escape response. We show that in the face of threat arthropods coordinate their behavior and cardiac activity in a rapid and flexible manner.

Polarized object detection in crabs: a two-channel system.

Many animal species take advantage of polarization vision for vital tasks such as orientation, communication and contrast enhancement. Previous studies have suggested that decapod crustaceans use a two-channel polarization system for contrast enhancement. Here, we characterize the polarization contrast sensitivity in a grapsid crab. We estimated the polarization contrast sensitivity of the animals by quantifying both their escape response and changes in heart rate when presented with polarized motion stimuli. The motion stimulus consisted of an expanding disk with an 82 deg polarization difference between the object and the background. More than 90% of animals responded by freezing or trying to avoid the polarized stimulus. In addition, we co-rotated the electric vector (e-vector) orientation of the light from the object and background by increments of 30 deg and found that the animals' escape response varied periodically with a 90 deg period. Maximum escape responses were obtained for object and background e-vectors near the vertical and horizontal orientations. Changes in cardiac response showed parallel results but also a minimum response when e-vectors of object and background were shifted by 45 deg with respect to the maxima. These results are consistent with an orthogonal receptor arrangement for the detection of polarized light, in which two channels are aligned with the vertical and horizontal orientations. It has been hypothesized that animals with object-based polarization vision rely on a two-channel detection system analogous to that of color processing in dichromats. Our results, obtained by systematically varying the e-vectors of object and background, provide strong empirical support for this theoretical model of polarized object detection.

New evidence on an old question: is the "fight or flight" stage present in the cardiac and respiratory regulation of decapod crustaceans?

The ability to stay alert to subtle changes in the environment and to freeze, fight or flight in the presence of predators requires integrating sensory information as well as triggering motor output to target tissues, both of which are associated with the autonomic nervous system. These reactions, which are commonly related to vertebrates, are the fundamental physiological responses that allow an animal to survive danger. The circulatory activity in vertebrates changes in opposite phases. The stage where circulatory activity is high is termed the "fight or flight stage", while the stage where circulatory activity slows down is termed the "rest and digest stage". It may be assumed that highly evolved invertebrates possess a comparable response system as they also require rapid cardiovascular and respiratory regulation to be primed when necessary. However, in invertebrates, the body plan may have developed such a system very differently. Since this topic is insufficiently studied, it is necessary to extend studies for a comparative analysis. In the present review, we use our own experimental results obtained in the crab Neohelice granulata and both older and newer findings obtained by other authors in decapod crustaceans as well as in other invertebrates, to compare the pattern of change in circulatory activity, especially in the "fight or flight" stage. We conclude that the main features of neuroautonomic regulation of the cardiac function were already present early in evolution, at least in highly evolved invertebrates, although conspicuous differences are also evident.

Characterization of the cardiac ganglion in the crab Neohelice granulata and immunohistochemical evidence of GABA-like extrinsic regulation.

The aim of the present work is to provide an anatomical description of the cardiac system in the crab Neohelice granulata and evidence of the presence of GABA by means of immunohistochemistry. The ganglionic trunk was found lying on the inner surface of the heart's dorsal wall. After dissection, this structure appeared as a Y-shaped figure with its major axis perpendicular to the major axis of the heart. Inside the cardiac ganglion, we identified four large neurons of 63.7 µm ± 3.7 in maximum diameter, which were similar to the motor neurons described in other decapods. All the GABA-like immunoreactivity (GABAi) was observed as processes entering mainly the ganglionic trunk and branching in slender varicose fibers, forming a network around the large neurons suggesting that GABAi processes contact them. Our findings strengthen previous results suggesting that the GABAergic system mediates the cardio-inhibitory response upon sensory stimulation.

Conditioning of an autonomic response in Crustacea.

Reports on experience-dependent changes in invertebrate autonomic function are few. In the crab Chasmagnathus, repeated presentations of a visual danger stimulus (VDS) elicit long-term cardiac adjustments. Although these changes can be explained in terms of an associative process, they do not necessarily indicate an anticipatory conditioned response. In the present work, we investigated anticipation of the cardio-inhibitory response (CIR) after classical conditioning. We found that an initially seemingly neutral stimulus, which could trigger only a brief CIR as part of an arousal/orienting response, following pairing with the unconditioned stimulus, 24 h after a second exposure, triggered a significantly stronger CIR response compared to controls. We propose that, as a result of training, the conditioned stimulus acquires a different biological meaning, allowing the crab to anticipate the aversive stimulus.

The cardiac response of the crab Chasmagnathus granulatus as an index of sensory perception.

When an animal's observable behavior remains unaltered, one can be misled in determining whether it is able to sense an environmental cue. By measuring an index of the internal state, additional information about perception may be obtained. We studied the cardiac response of the crab Chasmagnathus to different stimulus modalities: a light pulse, an air puff, virtual looming stimuli and a real visual danger stimulus. The first two did not trigger observable behavior, but the last two elicited a clear escape response. We examined the changes in heart rate upon sensory stimulation. Cardiac response and escape response latencies were also measured and compared during looming stimuli presentation. The cardiac parameters analyzed revealed significant changes (cardio-inhibitory responses) to all the stimuli investigated. We found a clear correlation between escape and cardiac response latencies to different looming stimuli. This study proved useful to examine the perceptual capacity independently of behavior. In addition, the correlation found between escape and cardiac responses support previous results which showed that in the face of impending danger the crab triggers several coordinated defensive reactions. The ability to escape predation or to be alerted to subtle changes in the environment in relation to autonomic control is associated with the complex ability to integrate sensory information as well as motor output to target tissues. This ;fear, fight or flight' response gives support to the idea of an autonomic-like reflexive control in crustaceans.

Cardiovascular component of the context signal memory in the crab Chasmagnathus.

Research on diverse models of memory in vertebrates demonstrates that behavioral, autonomic and endocrine responses occur together during fear conditioning. With invertebrates, no similar studies have been performed despite the extensive study of fear memory paradigms, as the context signal memory (CSM) of the crab Chasmagnathus granulatus, usually assessed by a behavioral parameter. Here, we study the crab's CSM, considering both the behavioral response and the concomitant neuroautonomic adjustments resulting in a heart rate alteration. Results show that upon the first presentation of the visual danger stimulus, a heart arrest followed by bradycardia is triggered together with a conspicuous escape response. The latter declines throughout training, while heart arrests become sporadic and bradycardia tends to deepen along the session. At test, 24 h after training, the outcome clearly contrasts with that shown at training, namely, stimulus presentation in the same context induces lower escape, no heart arrests and quick suppression of bradycardia. These results support the view that the same memory process brings about the changes in both responses. High escape, heart arrest and bradycardia are considered three parameters of the unconditioned response while minor escape, no heart arrests and bradycardia attenuation are three parameters of the learned response.

CS-US delay does not impair appetitive conditioning in Chasmagnathus.

Habituation and appetitive conditioning have been already described in the crab Chasmagnathus. The purpose of this work is to study whether associative learning can be obtained despite a long conditioned stimulus-unconditioned stimulus interval. Results of the first experiment show that the weakening of temporal contiguity does not prevent appetitive conditioning to occur while after a long 4-h delay, conditioning wanes completely. A second experiment was conducted, after one and three days of training respectively, confirming the above results. Though initially neutral the context trace may be still available immediately after training and for the period of two but not after 4:00 h, demonstrating a forward limit for the conditioning window. After 3 days of training, a further decrease in the exploratory activity suggested that a longer training could increase the relative weight of habituation. Conditioning and habituation seem to work as opponent processes in the crab CHASMAGNATHUS GRANULATUS: if habituation training in the box is followed by the administration of reinforcement after a short period of time, appetitive conditioning will take place. However, as this interval is increased, habituation prevails. A persistent effect of the exposure to a given environment that may underlie trace conditioning in this crab is discussed in adaptive terms.