Defense through sensory inactivation: sea hare ink reduces sensory and motor responses of spiny lobsters to food odors.

Antipredator defenses are ubiquitous and diverse. Ink secretion of sea hares (Aplysia) is an antipredator defense acting through the chemical senses of predators by different mechanisms. The most common mechanism is ink acting as an unpalatable repellent. Less common is ink secretion acting as a decoy (phagomimic) that misdirects predators' attacks. In this study, we tested another possible mechanism--sensory inactivation--in which ink inactivates the predator's reception of food odors associated with would-be prey. We tested this hypothesis using spiny lobsters, Panulirus argus, as model predators. Ink secretion is composed of two glandular products, one being opaline, a viscous substance containing concentrations of hundreds of millimolar of total free amino acids. Opaline sticks to antennules, mouthparts and other chemosensory appendages of lobsters, physically blocking access of food odors to the predator's chemosensors, or over-stimulating (short term) and adapting (long term) the chemosensors. We tested the sensory inactivation hypotheses by treating the antennules with opaline and mimics of its physical and/or chemical properties. We compared the effects of these treatments on responses to a food odor for chemoreceptor neurons in isolated antennules, as a measure of effect on chemosensory input, and for antennular motor responses of intact lobsters, as a measure of effect on chemically driven motor behavior. Our results indicate that opaline reduces the output of chemosensors by physically blocking reception of and response to food odors, and this has an impact on motor responses of lobsters. This is the first experimental demonstration of inactivation of peripheral sensors as an antipredatory defense.

Oesophageal chemoreceptors of blue crabs, Callinectes sapidus, sense chemical deterrents and can block ingestion of food.

Decapod crustaceans such as blue crabs possess a variety of chemoreceptors that control different stages of the feeding process. All these chemoreceptors are putative targets for feeding deterrents that cause animals to avoid or reject otherwise palatable food. As a first step towards characterizing the chemoreceptors that mediate the effect of deterrents, we used a behavioral approach to investigate their precise location. Data presented here demonstrate that chemoreceptors located on the antennules, pereiopods and mouthparts do not mediate the food-rejection effects of a variety of deterrents, both natural and artificial to crabs. Crabs always searched for deterrent-laced food and took it to their oral region. The deterrent effect was manifested as either rejection or extensive manipulation, but in both cases crabs bit the food. The biting behavior is relevant because the introduction of food into the oral cavity ensured that the deterrents gained access to the oesophageal taste receptors, and so we conclude that they are the ones mediating rejection. Additional support comes from the fact that a variety of deterrent compounds evoked oesophageal dilatation, which is mediated by oesophageal receptors and has been linked to food rejection. Further, there is a positive correlation between a compound's ability to elicit rejection and its ability to evoke oesophageal dilatation. The fact that deterrents do not act at a distance is in accordance with the limited solubility of most known feeding deterrents, and likely influences predator-prey interactions and their outcome: prey organisms will be attacked and bitten before deterrents become relevant.

Taste-mediated behavioral and electrophysiological responses by the predatory fish Ariopsis felis to deterrent pigments from Aplysia californica ink.

Chemical defenses are used by many organisms to avoid predation, and these defenses may function by stimulating predators' chemosensory systems. Our study examined detection mechanisms for components of defensive ink of sea hares, Aplysia californica, by predatory sea catfish, Ariopsis felis. Behavioral analyses show aplysioviolin and phycoerythrobilin are detected intra-orally and by barbels and are deterrent at concentrations as low as 0.1% full strength. We performed electrophysiological recordings from the facial-trigeminal nerve complex innervating the maxillary barbel and tested aplysioviolin, phycoerythrobilin, amino acids, and bile salts in cross-adaptation experiments. Amino acids and bile salts are known stimulatory compounds for teleost taste systems. Our results show aplysioviolin and phycoerythrobilin are equally stimulatory and completely cross-adapt to each other's responses. Adaptation to aplysioviolin or phycoerythrobilin reduced but did not eliminate responses to amino acids or bile salts. Adaptation to amino acids or bile salts incompletely reduced responses to aplysioviolin or phycoerythrobilin. The fact that cross-adaptations with aplysioviolin and phycoerythrobilin were not completely reciprocal indicates there are amino acid and bile salt sensitive fibers insensitive to aplysioviolin and phycoerythrobilin. These results indicate two gustatory pathways for aplysioviolin and phycoerythrobilin: one independent of amino acids and bile salts and another shared with some amino acids.

The neuroecology of chemical defenses.

Chemicals are a frequent means whereby organisms defend themselves against predators, competitors, parasites, microbes, and other potentially harmful organisms. Much progress has been made in understanding how a phylogenetic diversity of organisms living in a variety of environments uses chemical defenses. Chief among these advances is determining the molecular identity of defensive chemicals and the roles they play in shaping interactions between individuals. Some progress has been made in deciphering the molecular, cellular, and systems level mechanisms underlying these interactions, as well as how these interactions can lead to structuring of communities and even ecosystems. The neuroecological approach unifies practices and principles from these diverse disciplines and at all scales as it attempts to explain in a single conceptual framework the abundances of organisms and the distributions of species within natural habitats. This article explores the neuroecology of chemical defenses with a focus on aquatic organisms and environments. We review the concept of molecules of keystone significance, including examples of how saxitoxin and tetrodotoxin can shape the organization and dynamics of marine and riparian communities, respectively. We also describe the current status and future directions of a topic of interest to our research group-the use of ink by marine molluscs, especially sea hares, in their defense. We describe a diversity of molecules and mechanisms mediating the protective effects of sea hares' ink, including use as chemical defenses against predators and as alarm cues toward conspecifics, and postulate that some defensive molecules may function as molecules of keystone significance. Finally, we propose future directions for studying the neuroecology of the chemical defenses of sea hares and their molluscan relatives, the cephalopods.

Physiological and behavioral effects of chemoreceptors located in different body parts of the swimming crab Callinectes danae.

By perfusing their branchial chambers with filtered seawater, we have developed a preparation that allows us to maintain the swimming crab Callinectes danae outside water without any major effects on its cardiac activity. This in turn allowed us to selectively stimulate chemoreceptors located in different body parts, and specifically to discriminate between the receptors located in the branchial chambers and those located in the oral region (mainly in the mouthparts, antennules and antennae). We show that a taurine solution can evoke bradycardia when applied to the oral region or to a combination of the oral region and the branchial chambers. Although the precise localization of the oral region receptors involved remains to be determined, ablation experiments show that the olfactory organs (i.e., the antennules) are not involved. Finally, we show that although stimulating the pereiopods has no effect on the animals' cardiac activity it causes the animals to move, putatively to try to grasp a piece of food, a reaction not evoked by stimulating the gills or the oral regions. Our results lend support to the idea that chemoreceptors located in different parts of the body play different functional roles in decapod crustaceans.