Behavior Genetics

, Volume 41, Issue 5, pp 746–753

Gr39a, a Highly Diversified Gustatory Receptor in Drosophila, has a Role in Sexual Behavior

  • Kanako Watanabe
  • Gakuta Toba
  • Masayuki Koganezawa
  • Daisuke Yamamoto
Original Research


Sexual recognition among individuals is crucial for the reproduction of animals. In Drosophila, like in many other animals, pheromones are suggested to play an important role in conveying information about an individual, such as sex, maturity and mating status. Sex-specific cuticular hydrocarbon components are thought to be major sex pheromones in Drosophila, and are postulated to act through the gustatory system, since they are mostly non-volatile chemicals. However, very little is known about the molecular and neural bases of gustatory pheromone reception. So far, a few putative gustatory receptors, including Gr32a and Gr68a, have been implicated in courtship behavior. Here, we examine another putative gustatory receptor, Gr39a, which shares a cluster with both Gr32a and Gr68a in a molecular phylogeny of the gustatory receptor family, for its potential role in courtship behavior. The Gr39a gene produces four isoforms through alternative splicing of different 5′-most exons. A quantitative real-time PCR analysis showed that the expression levels of all four splice variants of Gr39a were reduced in a fly line in which a P element was inserted into the Gr39a locus. Homozygous and hemizygous males for the P-element insertion, as well as males in which Gr39a was knocked down by RNAi, all showed reduced courtship levels toward females. The courtship levels returned to normal when the P element was excised out. A close analysis of courtship behavior of the mutant males revealed that the average duration of a continuous courtship bout was significantly shorter in the mutants than in the wild type. The results suggest that Gr39a has a role in sustaining courtship behavior in males, possibly through the reception of a stimulating arrestant pheromone.


Drosophila Pheromone Gustatory receptor Sexual behavior Mutant 

Supplementary material

10519_2011_9461_MOESM1_ESM.doc (40 kb)
Supplementary material 1 (DOC 39 kb)
10519_2011_9461_MOESM2_ESM.eps (437 kb)
Fig. S1. Locomotor activity, and its relationship to courtship activity. (A) The locomotor activity of Gr39ak05106 mutant males was significantly lower than that of Canton-S males. Here, locomotor activity was defined as the number of times the fly crossed a reference line in the middle of an assay chamber in a 1-min observation period. Values are shown as the means ± SEM. Student’s t-test was used to evaluate the statistical significance of difference (***, P < 0.001). (B) Correlations between locomotor and courtship activities were examined for Canton-S wild-type and Gr39ak05106 mutant males. The locomotor activity of each individual was plotted against the CI of the same individual. No correlation was detected between the locomotor and courtship activities in both genotypes. Pearson’s correlation coefficients were as follows: Canton-S (r = -0.094), Gr39ak05106 (r = -0.218). The same data set of locomotor activities was used for both (A) and (B). In both (A) and (B), the numbers of males examined were as follows: Canton-S (n = 17), Gr39ak05106 (n = 27). (EPS 437 kb)


  1. Amrein H, Thorne N (2005) Gustatory perception and behavior in Drosophila melanogaster. Curr Biol 15:R673–R684PubMedCrossRefGoogle Scholar
  2. Antony C, Davis TL, Carlson DA, Pechiné JM, Jallon JM (1985) Compared behavioral responses of male Drosophila melanogaster (Canton S) to natural and synthetic aphrodisiacs. J Chem Ecol 11:1617–1629CrossRefGoogle Scholar
  3. Bellen HJ, Levis RW, Liao G, He Y, Carlson JW, Tsang G, Evans-Holm M, Hiesinger PR, Schulze KL, Rubin GM, Hoskins RA, Spradling AC (2004) The BDGP gene disruption project: single transposon insertions associated with 40% of Drosophila genes. Genetics 167:761–781PubMedCrossRefGoogle Scholar
  4. Bier E, Vaessin H, Shepherd S, Lee K, McCall K, Barbel S, Ackerman L, Carretto R, Uemura T, Grell E, Jan LY, Jan YN (1989) Searching for pattern and mutation in the Drosophila genome with a P-lacZ vector. Genes Dev 3:1273–1287PubMedCrossRefGoogle Scholar
  5. Billeter JC, Atallah J, Krupp JJ, Millar JG, Levine JD (2009) Specialized cells tag sexual and species identity in Drosophila melanogaster. Nature 461:987–991PubMedCrossRefGoogle Scholar
  6. Bray S, Amrein H (2003) A putative Drosophila pheromone receptor expressed in male-specific taste neurons is required for efficient courtship. Neuron 39:1019–1029PubMedCrossRefGoogle Scholar
  7. Brennan PA, Zufall F (2006) Pheromonal communication in vertebrates. Nature 444:308–315PubMedCrossRefGoogle Scholar
  8. Clyne PJ, Warr CG, Carlson JR (2000) Candidate taste receptors in Drosophila. Science 287:1830–1834PubMedCrossRefGoogle Scholar
  9. Cobb M, Jallon JM (1990) Pheromones, mate recognition and courtship stimulation in the Drosophila melanogaster species sub-group. Anim Behav 39:1058–1067CrossRefGoogle Scholar
  10. Coyne JA, Crittenden AP, Mah K (1994) Genetics of a pheromonal difference contributing to reproductive isolation in Drosophila. Science 265:1461–1464PubMedCrossRefGoogle Scholar
  11. Dahanukar A, Lei YT, Kwon JY, Carlson JR (2007) Two Gr genes underlie sugar reception in Drosophila. Neuron 56:503–516PubMedCrossRefGoogle Scholar
  12. Dietzl G, Chen D, Schnorrer F, Su KC, Barinova Y, Fellner M, Gasser B, Kinsey K, Oppel S, Scheiblauer S, Couto A, Marra V, Keleman K, Dickson BJ (2007) A genome-wide transgenic RNAi library for conditional gene inactivation in Drosophila. Nature 448:151–156PubMedCrossRefGoogle Scholar
  13. Dunipace L, Meister S, McNealy C, Amrein H (2001) Spatially restricted expression of candidate taste receptors in the Drosophila gustatory system. Curr Biol 11:822–835PubMedCrossRefGoogle Scholar
  14. Ejima A, Griffith LC (2008) Courtship initiation is stimulated by acoustic signals in Drosophila melanogaster. PLoS One 3:e3246PubMedCrossRefGoogle Scholar
  15. Ferveur JF (1997) The pheromonal role of cuticular hydrocarbons in Drosophila melanogaster. BioEssays 19:353–358PubMedCrossRefGoogle Scholar
  16. Ferveur JF (2005) Cuticular hydrocarbons: their evolution and roles in Drosophila pheromonal communication. Behav Genet 35:279–295PubMedCrossRefGoogle Scholar
  17. Ferveur JF, Sureau G (1996) Simultaneous influence on male courtship of stimulatory and inhibitory pheromones produced by live sex-mosaic Drosophila melanogaster. Proc R Soc Lond B Biol Sci 263:967–973CrossRefGoogle Scholar
  18. Gardiner A, Barker D, Butlin RK, Jordan WC, Ritchie MG (2008) Evolution of a complex locus: exon gain, loss and divergence at the Gr39a locus in Drosophila. PLoS One 3:e1513PubMedCrossRefGoogle Scholar
  19. Greenspan RJ (1995) Understanding the genetic construction of behavior. Sci Am 272:72–78PubMedCrossRefGoogle Scholar
  20. Haerty W, Jagadeeshan S, Kulathinal RJ, Wong A, Ravi Ram K, Sirot LK, Levesque L, Artieri CG, Wolfner MF, Civetta A, Singh RS (2007) Evolution in the fast lane: rapidly evolving sex-related genes in Drosophila. Genetics 177:1321–1335PubMedCrossRefGoogle Scholar
  21. Hall JC (1994) The mating of a fly. Science 264:1702–1714PubMedCrossRefGoogle Scholar
  22. Jallon JM (1984) A few chemical words exchanged by Drosophila during courtship and mating. Behav Genet 14:441–478PubMedCrossRefGoogle Scholar
  23. Kanzaki R, Ikeda A, Shibuya T (1994) Morphological and physiological properties of pheromone-triggered flipflopping descending interneurons of the male silkworm moth, Bombyx mori. J Comp Physiol A 175:1–14CrossRefGoogle Scholar
  24. Koganezawa M, Haba D, Matsuo T, Yamamoto D (2010) The shaping of male courtship posture by lateralized gustatory inputs to male-specific interneurons. Curr Biol 20:1–8PubMedCrossRefGoogle Scholar
  25. Krstic D, Boll W, Noll M (2009) Sensory integration regulating male courtship behavior in Drosophila. PLoS One 4:e4457PubMedCrossRefGoogle Scholar
  26. Lacaille F, Hiroi M, Twele R, Inoshita T, Umemoto D, Manière G, Marion-Poll F, Ozaki M, Francke W, Cobb M, Everaerts C, Tanimura T, Ferveur JF (2007) An inhibitory sex pheromone tastes bitter for Drosophila males. PLoS One 2:e661PubMedCrossRefGoogle Scholar
  27. Lee Y, Moon SJ, Montell C (2009) Multiple gustatory receptors required for the caffeine response in Drosophila. Proc Natl Acad Sci USA 106:4495–4500PubMedCrossRefGoogle Scholar
  28. Lin DM, Goodman CS (1994) Ectopic and increased expression of Fasciclin II alters motoneuron growth cone guidance. Neuron 13:507–523PubMedCrossRefGoogle Scholar
  29. Matsuo T (2008) Rapid evolution of two odorant-binding protein genes, obp57d and obp57e, in the Drosophila melanogaster species group. Genetics 178:1061–1072PubMedCrossRefGoogle Scholar
  30. Matsuo T, Sugaya S, Yasukawa J, Aigaki T, Fuyama Y (2007) Odorant-binding proteins Obp57d and Obp57e affect taste perception and host-plant preference in Drosophila sechellia. PLoS Biol 5:e118PubMedCrossRefGoogle Scholar
  31. Miyamoto T, Amrein H (2008) Suppression of male courtship by a Drosophila pheromone receptor. Nat Neurosci 11:874–876PubMedCrossRefGoogle Scholar
  32. Moon SJ, Köttgen M, Jiao Y, Xu H, Montell C (2006) A taste receptor required for the caffeine response in vivo. Curr Biol 16:1812–1817PubMedCrossRefGoogle Scholar
  33. Moon SJ, Lee Y, Jiao Y, Montell C (2009) A Drosophila gustatory receptor essential for aversive taste and inhibiting male-to-male courtship. Curr Biol 19:1623–1627PubMedCrossRefGoogle Scholar
  34. Park IK, Lee HS, Lee SG, Park JD, Ahn YJ (2000) Antifeeding activity of isoquinoline alkaloids identified in Coptis japonica roots against Hyphantria cunea (Lepidoptera: Arctiidae) and Agelastica coerulea (Coleoptera: Galerucinae). J Econ Entomol 93:331–335PubMedCrossRefGoogle Scholar
  35. Robertson HM, Preston CR, Phillis RW, Johnson-Schlitz DM, Benz WK, Engels WR (1988) A stable genomic source of P element transposase in Drosophila melanogaster. Genetics 118:461–470PubMedGoogle Scholar
  36. Robertson HM, Warr CG, Carlson JR (2003) Molecular evolution of the insect chemoreceptor gene superfamily in Drosophila melanogaster. Proc Natl Acad Sci USA 100(Suppl2):14537–14542PubMedCrossRefGoogle Scholar
  37. Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425PubMedGoogle Scholar
  38. Savarit F, Sureau G, Cobb M, Ferveur JF (1999) Genetic elimination of known pheromones reveals the fundamental chemical bases of mating and isolation in Drosophila. Proc Natl Acad Sci USA 96:9015–9020PubMedCrossRefGoogle Scholar
  39. Scott K, Brady R Jr, Cravchik A, Morozov P, Rzhetsky A, Zuker C, Axel R (2001) A chemosensory gene family encoding candidate gustatory and olfactory receptors in Drosophila. Cell 104:661–673PubMedCrossRefGoogle Scholar
  40. Sokolowski MB (2010) Social interactions in “simple” model systems. Neuron 65:780–794PubMedCrossRefGoogle Scholar
  41. Tsaur SC, Wu CI (1997) Positive selection and the molecular evolution of a gene of male reproduction, Acp26Aa of Drosophila. Mol Biol Evol 14:544–549PubMedGoogle Scholar
  42. Vosshall LB, Stocker RF (2007) Molecular architecture of smell and taste in Drosophila. Annu Rev Neurosci 30:505–533PubMedCrossRefGoogle Scholar
  43. Wyckoff GJ, Wang W, Wu CI (2000) Rapid evolution of male reproductive genes in the descent of man. Nature 403:304–309PubMedCrossRefGoogle Scholar
  44. Zawistowski S, Richmond RC (1986) Inhibition of courtship and mating of Drosophila melanogaster by the male-produced lipid, cis-vaccenyl acetate. J Insect Physiol 32:189–192CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Kanako Watanabe
    • 1
  • Gakuta Toba
    • 1
  • Masayuki Koganezawa
    • 1
  • Daisuke Yamamoto
    • 1
  1. 1.Division of Neurogenetics, Graduate School of Life SciencesTohoku UniversitySendaiJapan

Personalised recommendations