The synthetic substance hypoxanthine 3-N-oxide elicits alarm reactions in zebrafish (Danio rerio).

Zebrafish, one of the preferred study species of geneticists, is gaining increasing popularity in behavioral neuroscience. This small and prolific species may be an excellent tool with which the biological mechanisms of vertebrate brain function and behavior are investigated. Zebrafish has been proposed as a model organism in the analysis of fear responses and human anxiety disorders. Species-specific cues signaling the presence of predators have been successfully utilized in such research. Zebrafish has been shown to respond to its natural alarm substance with species-typical fear reactions. However, the extraction of this alarm substance and ascertaining its consistent dosing has been problematic. A synthetic substance with a known chemical identity and molecular weight would allow precise dosing and experimental control. Previously, the chemical component, hypoxanthine 3-N-oxide, common to several fish alarm substances has been identified and has been shown to elicit alarm reactions in fish species belonging to the Osteriophysan superorder. In the current study we investigate the effect of hypoxanthine 3-N-oxide by exposing zebrafish to three different concentrations of this synthetic substance. Our results show that the substance efficaciously induces species-typical fear reactions increasing the number of erratic movement episodes and jumps in zebrafish. We discuss the translational relevance of our findings and conclude that hypoxanthine 3-N-oxide will have utility to elicit fear responses in the laboratory in a precisely controlled manner in zebrafish.

Epidermal 'alarm substance' cells of fishes maintained by non-alarm functions: possible defence against pathogens, parasites and UVB radiation.

Many fishes possess specialized epidermal cells that are ruptured by the teeth of predators, thus reliably indicating the presence of an actively foraging predator. Understanding the evolution of these cells has intrigued evolutionary ecologists because the release of these alarm chemicals is not voluntary. Here, we show that predation pressure does not influence alarm cell production in fishes. Alarm cell production is stimulated by exposure to skin-penetrating pathogens (water moulds: Saprolegnia ferax and Saprolegnia parasitica), skin-penetrating parasites (larval trematodes: Teleorchis sp. and Uvulifer sp.) and correlated with exposure to UV radiation. Suppression of the immune system with environmentally relevant levels of Cd inhibits alarm cell production of fishes challenged with Saprolegnia. These data are the first evidence that alarm substance cells have an immune function against ubiquitous environmental challenges to epidermal integrity. Our results indicate that these specialized cells arose and are maintained by natural selection owing to selfish benefits unrelated to predator-prey interactions. Cell contents released when these cells are damaged in predator attacks have secondarily acquired an ecological role as alarm cues because selection favours receivers to detect and respond adaptively to public information about predation.

Nitrogen oxides elicit antipredator responses in juvenile channel catfish, but not in convict cichlids or rainbow trout: conservation of the ostariophysan alarm pheromone.

Recent studies with cyprinid and characin (superorder Ostariophysi) fishes suggest that purine-N-oxides function as chemical alarm cues (alarm pheromones) and that the nitrogen oxide functional group acts as the chief molecular trigger. To further test the hypothesis that the nitrogen-oxide functional group is evolutionarily conserved as an active component of the Ostariophysan alarm pheromone system, we exposed juvenile channel catfish (Ictalurus punctatus, Siluriformes) to conspecific skin extract, hypoxanthine-3-N-oxide (the putative alarm pheromone) and a suite of structurally and functionally similar compounds. Conspecific skin extract and hypoxanthine-3-N-oxide elicited significant increases in species typical antipredator behaviors. A structurally dissimilar compound possessing a nitrogen oxide functional group (pyridine-N-oxide) elicited a significant, but less intense alarm response. Compounds lacking a nitrogen oxide functional group were not significantly different from control stimuli. In addition, two non-Ostariophysan species known to possess chemical alarm cues (convict cichlids, Acrchocentrus nigrofasciatus, Cichlidae, Acanthopterygii and rainbow trout, Oncorhynchus mykiss, Salmonidae, Protacanthopterygii) did not show any increase in antipredator behavior in response to hypoxanthine-3-N-oxide. These data demonstrate the conservation of chemical alarm cues within at least three orders of the superorder Ostariophysi.

Multiple component reactions: an efficient nickel-catalyzed Reformatsky-type reaction and its application in the parallel synthesis of beta-amino carbonyl libraries.

Multiple-component condensations (MCC) where three or more reactants combine to afford a new core structure possessing the molecular features of its composite building blocks is a powerful method for the preparation of molecular diversity. We have developed an efficient, nickel-catalyzed, Reformatsky-type three-component condensation (3CC) reaction that affords beta-amino carbonyl compounds. The scope of the reaction is demonstrated both in the gram and microscale settings; 15 beta-amino esters, amides, and a ketone were prepared efficiently at the mmol scale, and a library of 64 beta-amino carbonyl compounds was generated at the micromol scale.