Novel natural ligands for Drosophila olfactory receptor neurones

SUMMARY Due to its well-defined genome, the fruitfly Drosophila melanogaster has become a very important model organism in olfactory research. Despite all the research invested, few natural odour ligands have been identified. By using a combined gas chromatographic—single receptor neurone recording technique (GC—SC), we set out to identify active odour molecules in head space-collected volatiles from preferred food sources, i.e. different overripe or rotting fruit. In total, we performed 101 GC—SC experiments on 85 contacted sensilla. Using GC—mass spectrometry, we identified 24 active compounds. Synthetic samples of these compounds were used to establish dose—response curves for several of the receptor neurone types encountered. The response patterns of individual neurones were repeatable, and neurones were found to reside in stereotyped pairs. In total, we identified eight distinct sensillum types based on response profiles of 12 olfactory receptor neurone types. In most recordings, a single GC peak would produce a strong response, whereas a few other, often chemically related, compounds would produce weaker responses. The GC—SC recordings revealed that the olfactory receptor neurones investigated were often selective and could be divided into distinct functional types with discrete characteristics. Dose—response investigations revealed very low response thresholds to the tested compounds. Six of the novel ligands were also tested for their behavioural effect in a T-maze set up. Of these, five elicited attraction and one elicited repulsion.

[1]  L. Vosshall Olfaction in Drosophila , 2000, Current Opinion in Neurobiology.

[2]  G. Gisselmann,et al.  Functional expression and characterization of a Drosophila odorant receptor in a heterologous cell system , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[3]  Sudhir Kumar,et al.  Comparative Genomics in Eukaryotes , 2005 .

[4]  S. Helfand,et al.  Isolation and characterization of an olfactory mutant in Drosophila with a chemically specific defect. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[5]  B. Hansson,et al.  Detection of fruit- and flower-emitted volatiles by olfactory receptor neurons in the polyphagous fruit chafer Pachnoda marginata (Coleoptera: Cetoniinae) , 2001, Journal of Comparative Physiology A.

[6]  John R. Carlson,et al.  A Novel Family of Divergent Seven-Transmembrane Proteins Candidate Odorant Receptors in Drosophila , 1999, Neuron.

[7]  D. Hubel Tungsten Microelectrode for Recording from Single Units. , 1957, Science.

[8]  H. Arn,et al.  The Electroantennographic Detector — a Selective and Sensitive Tool in the Gas Chromatographic Analysis of Insect Pheromones , 1975 .

[9]  Stephen M. Mount,et al.  The genome sequence of Drosophila melanogaster. , 2000, Science.

[10]  A. Chess,et al.  Identification of candidate Drosophila olfactory receptors from genomic DNA sequence. , 1999, Genomics.

[11]  L. Wadhams Coupled Gas Chromatography — Single Cell Recording: a New Technique for Use in the Analysis of Insect Pheromones , 1982 .

[12]  E. Alcorta Characterization of the electroantennogram in Drosophila melanogaster and its use for identifying olfactory capture and transduction mutants. , 1991, Journal of neurophysiology.

[13]  John R. Carlson,et al.  Odor Coding in the Drosophila Antenna , 2001, Neuron.

[14]  Peter J. Clyne,et al.  Odor Coding in a Model Olfactory Organ: TheDrosophila Maxillary Palp , 1999, The Journal of Neuroscience.

[15]  R. Steinbrecht,et al.  Atlas of olfactory organs of Drosophila melanogaster , 1999 .

[16]  A. Borg-Karlson,et al.  Identification of plant volatiles activating single receptor neurons in the pine weevil (Hylobius abietis) , 1997, Journal of Comparative Physiology A.

[17]  T. Liljefors,et al.  Bioisosteric Approach to Elucidation of Binding of the Acetate Group of a Moth Sex Pheromone Component to Its Receptor , 1997, Journal of Chemical Ecology.

[18]  B. Hansson,et al.  Green leaf volatile‐detecting olfactory receptor neurones display very high sensitivity and specificity in a scarab beetle , 1999 .

[19]  W. Quinn,et al.  Classical conditioning and retention in normal and mutantDrosophila melanogaster , 1985, Journal of Comparative Physiology A.

[20]  Andrey Rzhetsky,et al.  A Spatial Map of Olfactory Receptor Expression in the Drosophila Antenna , 1999, Cell.

[21]  B. Hansson,et al.  Plant‐odour‐specific receptor neurones on the antennae of female and male Spodoptera littoralis , 1995 .

[22]  Á. Acebes,et al.  Increasing the Number of Synapses Modifies Olfactory Perception in Drosophila , 2001, The Journal of Neuroscience.

[23]  K. Störtkuhl,et al.  Functional analysis of an olfactory receptor in Drosophila melanogaster , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[24]  R. Stocker Drosophila as a focus in olfactory research: Mapping of olfactory sensilla by fine structure, odor specificity, odorant receptor expression, and central connectivity , 2001, Microscopy research and technique.

[25]  L. Vosshall,et al.  The molecular logic of olfaction in Drosophila. , 2001, Chemical senses.