1 1 Symposium Chemosensory Receptors Satellite DEVELOPMENT ...

285 Poster Central Olfaction and Chemical EcologyCHEMICALS RELEASED BY INJURED OR DISTURBEDCONSPECIFICS MEDIATE DEFENSIVE BEHAVIORS VIATHE AESTHETASC PATHWAY IN THE SPINY LOBSTERPANULIRUS ARGUSShabani S. 1 , Kamio M. 1 , Derby C. 1 1 Biology, Georgia State University,Atlanta, GAChemicals released by either injured or disturbed individuals inducefrom conspecifics a repertoire of defensive behaviors similar to whenattacked by predators. This has been demonstrated for many species,including crustaceans such as hermit crabs and crayfish. In this study,we demonstrate that the spiny lobster Panulirus argus also haschemically induced alarm responses, and furthermore demonstrate thesite of release and site of reception of the alarm cues. Alarm cues ofspiny lobsters are present in the blood (hemolymph) of injured animalsand in the urine of lobsters under simulated predatory attack (disturbedanimals). Hemolymph and urine activate defensive behaviors such asmoving away from the odor source, defensive movement of the 2ndantennae, shaking, and moving into a shelter, as well as suppressingfood-evoked searching. The alarm cues are species specific: Caribbeanspiny lobsters (Panulirus argus) show much greater alarm response tohemolymph from P. argus than to hemolymph from California spinylobsters (Panulirus interruptus) or blue crabs (Callinectes sapidus).Removal of the aesthetascs lead to reversal of behaviors towardhemolymph and urine, switching from defensive to attractive behaviors.In conclusion, we have identified the types of defensive behaviorsproduced by, the source of, and site of reception of the alarm cues inspiny lobsters. Future goals include identifying the molecules involved,the cellular specificity of the responsive olfactory neurons, and theecological context of the alarm response. Supported by NSF IBN-0324435, 9876754, 0322773286 Poster Central Olfaction and Chemical EcologyEVIDENCE THAT A VOLATILE MOLECULE, 3-DECANOL, INHERMIT CRAB BLOOD SIGNALS SHELL AVAILABILITY TOCONSPECIFICSRittschof D. 1 , Schmidt G. 2 , Harder T. 2 1 Duke University, Beaufort, NC;2 Institute for Chemistry and Biology of the Marine Environment(ICBM), University of Oldenburg, Oldenburg, GermanyHermit crabs in poor fitting shells are attracted to and obtaingastropod shells at gastropod and hermit crab predation sites. Hermitcrab hemolymph attracts conspecifics within minutes. We hypothesizedthat aquatic hermit crabs might use volatile molecules in hemolymph asdo land hermit crabs (Thacker, 1994, Small and Thacker, 1994).Volatiles from, sea water, hemolymph of hermit crabs Clibanariusvittatus and Pagurus pollicaris and several brachyuran crabs werepurged from a water-hemolymph solution, trapped in seawater andtested for induction of shell-investigation behavior in shell investigationbioassays. Volatiles extracted from crustacean hemolymph byheadspace solid-phase microextraction (SPME) were analysed bycoupled gas-chromatography mass-spectrometry (GC-MS). Twoalcohols, 3-decanol, present in C. vittatus hemolymph, and 2-ethyl-1-hexanol, present in hemolymph of all tested crustacean species, weretested in shell investigation bioassays. 3-decanol stimulated shellinvestigationbehavior in conspecifics, while 2-ethyl-1-hexanol did not.The response of C. vittatus to this volatile water soluble moleuclessupports the hypothesis that detection and response to volatile cuesevolved before the transition of hermit crabs to land.287 Poster Central Olfaction and Chemical EcologyN-ACETYLGLUCOSAMINO-1,5-LACTONE IS A CANDIDATESEX PHEROMONE IN FEMALE BLUE CRABSKamio M. 1 , Kubanek J. 2 , Derby C. 1 1 Biology, Georgia State University,Atlanta, GA; 2 Biology, Georgia Institute of Technology, Atlanta, GAPremolt female blue crabs (Callinectes sapidus) release a sexpheromone in their urine. Males detect this pheromone using antennularsensors, resulting in mating behaviors that include precopulatorydisplay and grabbing and guarding females. Male crabs also release asex pheromone that attracts premolt females. The molecular identity ofthese pheromones remains unknown. The goal of our study is to identifythese molecules using bioassay guided fractionation and analysis ofdifferences in the composition of male and female urine. In search ofsex pheromones, we found differences in the chemical composition ofmale and female urine using NMR spectral analysis (Kamio et al.,AChemS 2004 abstract #239). One compound that is specific to premoltfemales was purified by ultrafiltration and HPLC. NMR spectralanalysis led to the characterization of this candidate female sexpheromone as N-acetylglucosamino-1, 5-lactone (NAGL), which ismore abundant in urine of premolt females than in urine of eitherintermolt, premolt, or postmolt males or intermolt or postmolt females.NAGL, which is an oxidized form of N-acetylglucosamine, has neverbefore been reported as a natural product, although synthetic NAGL hasbeen reported to inhibit chitobiase of bacteria and vertebrates, whichdegrades chitin. Preliminary behavioral experiments demonstrated thatmale crabs can detect NAGL. Further behavioral testing is planned toevaluate the possible role of NAGL and other minor metabolites as sexpheromones. Supported by the NSF Grants IBN-9876754 and IBN-0322773 to the Center for Behavioral Neuroscience and Japan Societyfor the Promotion of Science Postdoctoral Fellowship for ResearchAbroad (to M.K.).288 Poster Central Olfaction and Chemical EcologyISOLATION AND STRUCTURE ELUCIDATION OF THE SEALAMPREY MIGRATORY PHEROMONEDvornikovs V. 1 , Fine J.M. 2 , Hoye T.R. 1 , Jeffrey C.S. 1 , Shao F. 1 , WangJ. 1 , Vrieze L.A. 2 , Anderson K.R. 1 , Sorensen P.W. 2 1 Department ofChemistry, University of Minnesota, Minneapolis, MN; 2 Department ofFisheries & Wildlife, University of Minnesota, St. Paul, MNThe sea lamprey (Petromyzon marinus) is one of the most ancientvertebrates and has a well-developed olfactory system. It begins life infreshwater streams, which it then leaves to parasitize other fishes beforeeventually maturing and returning to spawn. Migratory lampreyrecognize suitable spawning streams using a pheromone produced bystream-resident, conspecific larvae. The pheromone contains two newand one known [the lamprey bile acid, petromyzonol sulfate (PS)]active components. Here we report the structure elucidation of the newcompounds and aspects of their biological activity. The most potent,petromyzonamine disulfate (PADS), is a novel disulfated aminosterolstructurally related to squalamine, an antibiotic produced by dogfishshark. Lamprey were attracted to isolated (or synthetic) PADS atconcentrations down to 10 -13 M. The second new component,petromyzosterol disulfate (PSDS, attractive at 10 -12 M), was found to bea sulfated ergostadienetriol derivative. The biological activity of variousmixtures of PADS, PSDS, and PS confirmed that they synergize oneanother and comprise the majority of the cue. Use of syntheticpheromone components is planned for controlling populations in theGreat Lakes, where the sea lamprey is an invasive pest. This multicomponentcue is the first migratory pheromone to be identified in afish. Funded by the Great Lakes Fishery Commission and the NationalInstitutes of Health (GM65597).72