Abstracts - Association for Chemoreception Sciences

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P O S T E R S been isolated in P. shermani. The DNA sequences clustered into 3 subfamilies that are less than 80% similar in sequence identity. In situ hybridization (ISH) using RNA probes revealed 2 different patterns of V2R RNA expression in the VNO. In one pattern, the number of cells labeled by the RNA probe was positively correlated to the volume of the VNO. In other words, the density of cells expressing a particular class of V2R was similar across individuals. In the other pattern, the number of labeled cells was unrelated to the volume of the VNO. In this case, individuals with a larger VNO had proportionally fewer labeled cells than did individuals with a smaller VNO. Thus, individual differences in V2R expression may contribute to differences in sensory function. Since males typically have a larger VNO than females, both in absolute size and relative to body size, this may translate into sex differences in responses to sensory information. Acknowledgements: NSF IOS-0818554 to LDH, NSF IOS- 0808589 to KMK and LDH, NSF predoctoral fellowship to KMK. #P157 POSTER SESSION IV: CHEMOSENSORY TRANSDUCTION AND SIGNALING Molecular characterization and localization of olfactoryspecific ionotropic glutamate receptors in lobster olfactory receptor neurons Elizabeth A Corey 1 , Yuriy Bobkov 1 , Barry W Ache 1,2 1 Whitney Laboratory, Center for Smell and Taste, and McKnight Brain Institute St Augustine, FL, USA, 2 Depts. of Biology and Neuroscience, University of Florida Gainesville, FL, USA The molecular basis of olfaction in crustaceans, a major group of arthropods, is still uncertain. While there is accumulating evidence that G protein activation of metabotropic signaling pathways is involved in crustacean olfactory signal transduction, a lobster olfactory-specific ionotropic glutamate receptor, OET07, appears to be an ortholog of the recently discovered Drosophila olfactory variant of ionotropic glutamate receptors (IRs). These results suggest that crustacean olfactory transduction mechanisms are at least in part similar to those of insects. As a first step towards understanding the role of IR-mediated signaling in crustacean olfaction, we have begun to characterize the expression of lobster olfactory IRs. We have cloned two full length lobster IR orthologs, including that of OET07, as well as partial sequences from other additional potential IRs, and demonstrated that all of the putative IRs can be detected in lobster olfactory tissue by RT-PCR. The lobster ortholog of OET07 can be detected in most, if not all, olfactory receptor neurons by in situ hybridization, and can be localized to the transduction compartment (outer dendrites) by western blot and immunocytochemistry. These results support a role for IR-mediated signaling in the lobster olfactory transduction mechanism. Using heterologous expression, we are currently attempting to determine whether the lobster IR orthologs can function as ionotropic receptors. Acknowledgements: Supported by grants from the National Institute on Deafness and Other Communication Disorders. #P158 POSTER SESSION IV: CHEMOSENSORY TRANSDUCTION AND SIGNALING Measuring Ensemble Activity in Lobster ORNs through Calcium Imaging Yuriy V. Bobkov 1 , Kirill Y. Ukhanov 1 , Ill Park 3 , Jose C. Principe 3 , Barry W. Ache 1,2 1 Whitney Laboratory, Center for Smell and Taste, and McKnight Brain Institute, University of Florida Gainesville, FL, USA, 2 Depts. of Biology and Neuroscience, University of Florida Gainesville, FL, USA, 3 Dept. of Electrical and Computer Engineering, University of Florida Gainesville, FL, USA Lobster ORNs can be imaged in the olfactory organ in situ, thereby maintaining the normal polarity of the cells and the ionic environment of the olfactory cilia. The preparation gives simultaneous access to hundreds of ORNs that are viable for hours, thereby allowing rigorous characterization of their steadystate and dynamic properties. Odorants change the level of cytoplasmic Ca 2+ in a dose-dependent manner in ORNs loaded with Ca 2+ -sensitive indicator either through bath application or via a patch electrode. The kinetics and amplitude of the odorantevoked Ca 2+ signal correlate with the excitatory inward current, the degree of membrane depolarization, and the number of evoked action potentials, thereby establishing the physiological relevance of the Ca 2+ signal. Spontaneous periodic Ca 2+ transients in many ORNs correlate with spontaneous bursts of action potentials measured in single cells in the same cluster. We are using signal processing algorithms to analyze the level of correlated activity between these ORNs and the extent to which periodic calcium oscillations in different ORNs are synchronized by common intermittent excitatory input to test the predictions of our computational model for ensemble burst coding in these cells and the potential relevance of bursting input to olfactory scene analysis. Acknowledgements: Supported by the NIDCD (DC001655, DC005995) #P159 POSTER SESSION IV: CHEMOSENSORY TRANSDUCTION AND SIGNALING Ca Imaging of Response Properties of Olfactory Receptor Neurons of Spiny Lobsters, Panulirus argus Manfred Schmidt, Tizeta Tadesse, Charles D Derby Neuroscience Institute, Georgia State University Atlanta, GA, USA The spiny lobster, Panulirus argus, is an established model for studying olfaction. Its olfactory organ consists of tufts of specialized sensilla – aesthetascs – on the lateral flagella of the antennules with each aesthetasc containing a cluster of > 300 olfactory receptor neurons (ORNs). Although transduction mechanisms in aesthetasc ORNs have been analyzed in detail with patch clamp electrophysiology (Ache and Young, Neuron 48:417- 430, 2005), very little is known about their basic response properties such as sensitivity and spectral tuning. To study these questions, we established Ca imaging of ORN responses in an in vitro ‘slice’ preparation similar to that used for patch clamp recordings. Short segments of lateral flagella were incubated in the Ca indicator Fluo-4 AM, mounted in an experimental chamber perfused with Panulirus saline, and imaged with an epifluorescence microscope equipped with monochromator and CCD camera. Our initial results showed that within each cluster several ORNs took up Fluo-4 and responded with a robust increase in fluorescence to global depolarization with high K + 80 | AChemS Abstracts 2010 Abstracts are printed as submitted by the author(s)
saline. Some of the labeled ORNs showed spontaneous rhythmic changes in fluorescence intensity. Puffing mixtures of food-related chemicals onto the aesthetascs with a multibarrel pipette caused a stimulus-coupled and transient increase in fluorescence intensity in some ORNs and a decrease in fluorescence intensity in others. Based on previous electrophysiological results (Michel et al., J.Neurophys. 65:446-453, 1991; Bobkov and Ache, J.Neurophys. 97:1052-1057, 2007), we interpret the rhythmic changes in fluorescent intensity as representing spontaneous bursting activity and the transient increases or decreases in fluorescent intensity as representing excitatory or inhibitory responses in ORN firing. Acknowledgements: Supported by NIH grant DC00312 and a GSU Brains & Behavior grant #P160 POSTER SESSION IV: CHEMOSENSORY TRANSDUCTION AND SIGNALING Evolution of haematophagy: what one moth species can teach us Sharon R. Hill 1 , Jennifer Zaspel 2 , Bill S. Hansson 3 , Susan Weller 2 , Rickard Ignell 1 1 Division of Chemical Ecology, Department of Plant Protection Biology, Swedish Agricultural University Alnarp, Sweden, 2 Department of Entomology, University of Minnesota St. Paul, MN, USA, 3 Department of Evolutionary Neuroethology, Max Planck Institute for Chemical Ecology Jena, Germany The vampire moth, Calyptra thalictri, provides an interesting model through which to study the evolution of haematophagic behavior. Contrary to the well-characterized model systems in mosquitoes, blood feeding in this moth appears to be a trait recently acquired. The morphological and physiological evidence we present supports the hypothesis that, in blood feeding male C. thalictri, a reduction in the number of antennal sensilla coeloconica shown to respond to vertebrate host-related compounds appears to result in the ability to overcome behavioral repulsion and/or acquire attraction to vertebrates, thus providing a new opportunity for these males to land on and feed from vertebrate hosts. Male moths were collected from two sites north of Vladivostok in Far Eastern Russia and from one site in Rotskär, Sweden. The antennae of these moths were examined using scanning electron microscopy, which revealed a lower number of sensilla coeloconica in blood feeding compared to non-blood feeding males. The response spectra of these sensilla were described using the single sensillum recording technique. These sensilla responded to a variety of vertebrate-related volatile compounds such as short chain carboxylic acids, phenolics, ketones and aldehydes, as well as ammonia. Acknowledgements: This work was supported by the Linnaeus-program Insect Chemical Ecology, Ethology and Evolution IC-E3. #P161 POSTER SESSION IV: CHEMOSENSORY TRANSDUCTION AND SIGNALING Sex Pheromone Receptor Specificity in the European Corn Borer Moth, Ostrinia nubilalis Kevin W Wanner 1 , Andrew S Nichols 2 , Jean E Allen 1 , Peggy L Bunger 1 , Stephen F Garczynski 3 , Charles E Linn 4 , Hugh M Robertson 5 , Charles W Luetje 2 1 Montana State University Bozeman, MT, USA, 2 University of Miami Miami, FL, USA, 3 USDA-ARS Wapato, WA, USA, 4 Cornell University Ithica, NY, USA, 5 University of Illinois Urbana-Champaign Urbana, IL, USA Female moths (Order Lepidoptera) produce and release a mixture of related fatty acid derivatives from their pheromone gland to which males respond from long distances. In many cases, subtle changes in carbon chain length, double bond location and isomer blend differentiate the pheromones of closely related species. The origin and mechanism of the variation in male detection that enables the evolution of new pheromone blends is not known. The European corn borer is used as a model system to study the evolution of sex pheromones among closely related races and species. It exists as two separate sex pheromone races: ECB(Z) females produce a 97:3 blend of Z11- and E11-tetradecenyl acetate whereas ECB(E) females produce an opposite 1:99 ratio of the Z and E isomers. Males of each race respond specifically to their conspecific female’s blend. The Asian corn borer, a closely related species, uses a 3:2 blend of Z12- and E12-tetradecenyl acetate. We used homology-dependent (degenerate PCR primers designed to conserved amino acid motifs) and homology-independent (pyrophosphate sequencing of antennal cDNA) approaches to identify five candidate sex pheromone transcripts (OnOr1 & 3-6) from ECB(Z). OnOr1 & 3-6 were expressed 14-100 times more abundantly in male compared to female antennae. OnOr6, characterized in Xenopus oocyets, was highly selective for Z11- tetradecenyl acetate (EC50 = 0.86 +/- 0.27 microM). OnOr6 was 1000 times less sensitive to the E11 isomer. Surprisingly, OnOr1, 3 and 5 responded more broadly to all four pheromones tested (Z11, E11, Z12 and E12 components). Receptors broadlyresponsive to a class of pheromone components may provide a mechanism for variation in the male moth response that enables population level shifts in pheromone blend use. #P162 POSTER SESSION IV: CHEMOSENSORY TRANSDUCTION AND SIGNALING Molecular characterization of accessory proteins mediating sexual selection in two Ostrinia species Jean E. Allen, Kevin W. Wanner Montana State University Bozeman, MT, USA Sexual selection and mating in moths is mediated by olfactory sensing of pheromone blends. Differences between pheromone blends are detected at the periphery of the olfactory system by receptors and accessory proteins expressed in trichoid sensilla on the male antennae. Pheromone binding proteins (PBPs) and sensory neuron membrane proteins (SNMPs) are involved in pheromone detection, and may play a role in discrimination of pheromone blends. The European corn borer (Ostrinia nubilalis) (ECB) exists as two different races, the Z-race uses (Z)-11- tetradecenyl acetate as the main component of its pheromone blend while the E-race uses (E)-11-tetradecenyl acetate. The closely related Asian corn borer (Ostrinia furnacalis) (ACB) uses a slightly different pheromone, (E) and (Z)-12-tetradecenyl acetate. We hypothesized that changes in the pheromone components are accompanied by changes in the sequence or expression level of the genes involved in their detection. Partial transcripts of 5 PBPs and 2 SNMPs from the ECB Z-race were identified by pyrosequencing antennal cDNA. Complete cDNA sequences were obtained by rapid amplification of cDNA ends. Primers designed to untranslated regions were used to amplify open reading frames from both ECB races and ACB. Evidence of positive selection within predicted ligand-binding sites was analyzed by calculating the ratio of synonymous and non synonymous substitutions. Expression levels of PBPs and SNMPs in the antennae were measured using quantitative real-time PCR. We found that there was no evidence for positive selection in the sequences or expression levels of the PBPs and SNMPs, and P O S T E R S Abstracts are printed as submitted by the author(s) Abstracts | 81
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AChemS Association for Chemorecepti
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AChemS Association for Chemorecepti
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We are pleased to announce that sev
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#1 GIVAUDAN LECTURE Normal and Canc
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and against delta ENaC is being pur
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#13 SYMPOSIUM - GENETICS OF HUMAN O
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#19 SYMPOSIUM - CILIA, SENSORY DYSF
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#26 PLATFORM PRESENTATIONS - POLAK
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esponse selectivity for both OSNs a
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elicit glucagon-like peptide-1 (GLP
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contrast, changing the concentratio
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of OBPs, we have systematically sup
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whereas mouse and joystick reaction
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POSTER PRESENTATIONS #P1 POSTER SES
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Page 63 and 64: gene, HMBS was used. All primers we
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seem to indicate that total nasal v
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possibly suggesting non-apoptotic m
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#P325 POSTER SESSION VII: OLFACTORY
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of 7/10, and causing eyes to water
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showed that N1/P2 latencies were lo
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differentiated patients with AD (M=
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pleasantness 3)of mixtures (A,B), A
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Index Aarts, H - P328 Abe, K - P88,
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Haase, L - P3, P4, P29, P355 Haddad
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Murata, Y - P272 Murphy, C - P3, P4
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Veldhuizen, M - P7, P24, P25, P242,
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Registration 7:30 am to 1:00 pm, 6:
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See you next year! AChemS 33rd Annu
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Abstracts - Association for Chemoreception Sciences

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