A new Drosophila melanogaster fly that expresses GFP-tagged Orco
Insects perceive countless odors with the help of odorant receptors (ORs) expressed by olfactory sensory neurons (OSNs). OR complexes are composed of odor-specific proteins OrX and olfactory co-receptor proteins Orco, with both extracellular C-terminus and intracellular N-terminus. Here, we generate...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-05-01
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| Series: | Frontiers in Ecology and Evolution |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fevo.2023.1150532/full |
| _version_ | 1797828936300756992 |
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| author | Kalpana Jain Regina Stieber Sabine Kaltofen Bill S. Hansson Dieter Wicher |
| author_facet | Kalpana Jain Regina Stieber Sabine Kaltofen Bill S. Hansson Dieter Wicher |
| author_sort | Kalpana Jain |
| collection | DOAJ |
| description | Insects perceive countless odors with the help of odorant receptors (ORs) expressed by olfactory sensory neurons (OSNs). OR complexes are composed of odor-specific proteins OrX and olfactory co-receptor proteins Orco, with both extracellular C-terminus and intracellular N-terminus. Here, we generated a genetically modified Drosophila melanogaster fly line that expresses Orco proteins with an N-terminally fused green fluorescent protein (GFP). We generated four genotypes (homozygous and heterozygous) for which we performed antennal cryosectioning and immunolabeling to determine which genotype would show high GFP labeling intensity in the dendritic areas of OSNs. We found that fluorescent intensity was the highest in homozygous flies. As Orco is known to guide OrX proteins to the OSN dendrites, we checked whether the fusion of GFP to the Orco N-terminus may affect such trafficking and thereby OR function. Using single sensillum recording (SSR), we found no difference in the olfactory capability of homozygous flies compared with wild-type flies. As future application, we want to use this transgenic fly line to resolve the structure of OR complexes using Cryo electron microscopy (cryo-EM). |
| first_indexed | 2024-04-09T13:12:23Z |
| format | Article |
| id | doaj.art-72cafa6c49fe4ada9650280d5e0b67e5 |
| institution | Directory Open Access Journal |
| issn | 2296-701X |
| language | English |
| last_indexed | 2024-04-09T13:12:23Z |
| publishDate | 2023-05-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Ecology and Evolution |
| spelling | doaj.art-72cafa6c49fe4ada9650280d5e0b67e52023-05-12T06:23:29ZengFrontiers Media S.A.Frontiers in Ecology and Evolution2296-701X2023-05-011110.3389/fevo.2023.11505321150532A new Drosophila melanogaster fly that expresses GFP-tagged OrcoKalpana JainRegina StieberSabine KaltofenBill S. HanssonDieter WicherInsects perceive countless odors with the help of odorant receptors (ORs) expressed by olfactory sensory neurons (OSNs). OR complexes are composed of odor-specific proteins OrX and olfactory co-receptor proteins Orco, with both extracellular C-terminus and intracellular N-terminus. Here, we generated a genetically modified Drosophila melanogaster fly line that expresses Orco proteins with an N-terminally fused green fluorescent protein (GFP). We generated four genotypes (homozygous and heterozygous) for which we performed antennal cryosectioning and immunolabeling to determine which genotype would show high GFP labeling intensity in the dendritic areas of OSNs. We found that fluorescent intensity was the highest in homozygous flies. As Orco is known to guide OrX proteins to the OSN dendrites, we checked whether the fusion of GFP to the Orco N-terminus may affect such trafficking and thereby OR function. Using single sensillum recording (SSR), we found no difference in the olfactory capability of homozygous flies compared with wild-type flies. As future application, we want to use this transgenic fly line to resolve the structure of OR complexes using Cryo electron microscopy (cryo-EM).https://www.frontiersin.org/articles/10.3389/fevo.2023.1150532/fullinsect olfactionolfactory sensory neuronantennaimmunohistochemistrysingle sensillum recordingDrosophila melanogaster |
| spellingShingle | Kalpana Jain Regina Stieber Sabine Kaltofen Bill S. Hansson Dieter Wicher A new Drosophila melanogaster fly that expresses GFP-tagged Orco Frontiers in Ecology and Evolution insect olfaction olfactory sensory neuron antenna immunohistochemistry single sensillum recording Drosophila melanogaster |
| title | A new Drosophila melanogaster fly that expresses GFP-tagged Orco |
| title_full | A new Drosophila melanogaster fly that expresses GFP-tagged Orco |
| title_fullStr | A new Drosophila melanogaster fly that expresses GFP-tagged Orco |
| title_full_unstemmed | A new Drosophila melanogaster fly that expresses GFP-tagged Orco |
| title_short | A new Drosophila melanogaster fly that expresses GFP-tagged Orco |
| title_sort | new drosophila melanogaster fly that expresses gfp tagged orco |
| topic | insect olfaction olfactory sensory neuron antenna immunohistochemistry single sensillum recording Drosophila melanogaster |
| url | https://www.frontiersin.org/articles/10.3389/fevo.2023.1150532/full |
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