Behavioral analysis of Drosophila transformants expressing human taste receptor genes in the gustatory receptor neurons.
Transgenic Drosophila expressing human T2R4 and T2R38 bitter-taste receptors or PKD2L1 sour-taste receptor in the fly gustatory receptor neurons and other tissues were prepared using conventional Gal4/UAS binary system. Molecular analysis showed that the transgene mRNAs are expressed according to the tissue specificity of the Gal4 drivers. Transformants expressing the transgene taste receptors in the fly taste neurons were then studied by a behavioral assay to analyze whether transgene chemoreceptors are functional and coupled to the cell response. Since wild-type flies show strong aversion against the T2R ligands as in mammals, the authors analyzed the transformants where the transgenes are expressed in the fly sugar receptor neurons so that they promote feeding ligand-dependently if they are functional and activate the neurons. Although the feeding preference varied considerably among different strains and individuals, statistical analysis using large numbers of transformants indicated that transformants expressing T2R4 showed a small but significant increase in the preference for denatonium and quinine, the T2R4 ligands, as compared to the control flies, whereas transformants expressing T2R38 did not. Similarly, transformants expressing T2R38 and PKD2L1 also showed a similar preference increase for T2R38-specific ligand phenylthiocarbamide (PTC) and a sour-taste ligand, citric acid, respectively. Taken together, the transformants expressing mammalian taste receptors showed a small but significant increase in the feeding preference that is taste receptor and also ligand dependent. Although future improvements are required to attain performance comparable to the endogenous robust response, Drosophila taste neurons may serve as a potential in vivo heterologous expression system for analyzing chemoreceptor function.
Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidori-Amamiyacho,Aoba-ku, Sendai, Japan.
Ryota
R
Adachi
Yuko
Y
Sasaki
Hiromi
H
Morita
Michio
M
Komai
Hitoshi
H
Shirakawa
Tomoko
T
Goto
Akira
A
Furuyama
Kunio
K
Isono
D000818:Animals; D030801:Animals, Genetically Modified; D015220:Calcium Channels / Q000235:genetics / Q000378:metabolism; D004330:Drosophila; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism; D005518:Food Preferences; D005786:Gene Expression Regulation / Q000235:genetics*; D006801:Humans; D012333:RNA, Messenger; D011956:Receptors, Cell Surface / Q000235:genetics / Q000378:metabolism*; D043562:Receptors, G-Protein-Coupled / Q000235:genetics / Q000378:metabolism; D011984:Sensory Receptor Cells / Q000502:physiology*; D013649:Taste / Q000235:genetics*; D013650:Taste Buds / Q000166:cytology* / Q000502:physiology; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D000098706:Taste Receptors, Type 2
D015220:Calcium Channels; D029721:Drosophila Proteins; C557617:GAL4 protein, Drosophila; C117185:PKD2L1 protein, human; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D043562:Receptors, G-Protein-Coupled; D014157:Transcription Factors; C405956:gustatory receptor, Drosophila; D000098706:Taste Receptors, Type 2