Robust olfactory responses in the absence of odorant binding proteins
- Yale University, United States
Figures
Figure 1 with 1 supplement
Organization of the Drosophila antenna and morphology of mutant sensilla.
(A) Fly head. Arrow indicates antenna. Adapted from Menuz et al. (2014), with thanks to Dr Rudi Turner. (B) Large basiconic sensilla of control (left) and of an Obp19a-; Obp28a-; Obp83a-,b- mutant (right). Scale bar is 0.5 μm and also applies to (C). (C) Thin basiconic sensilla of control (left) and an Obp19a-; Obp28a-; Obp83a-,b- mutant (right). (D) Olfactory sensillum. Adapted from Larter et al. (2016) and Shanbhag et al. (1999). E = Epidermal cells. (E) Obp-to-sensillum map of the 10 functional types of basiconic sensilla.
Figure 1—figure supplement 1
Sensilla of mutant and control animals appear similar.
(A) Large basiconic sensilla are indicated by arrows. (B) Thin basiconic sensilla. Scale bars = 1 μm.
Figure 2 with 2 supplements
Responses of individual ORNs in each type of sensillum after removal of abundant Obps.
(A–F) ab1-ab6. Responses are to 0.5 s pulses of strong ligands. (SEM; n = 6). No significant differences were found by the Mann-Whitney U test.
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Figure 2—source data 1
Source data for spike numbers in Figure 2.
- https://doi.org/10.7554/eLife.51040.009
Figure 2—figure supplement 1
Hydrophobicity of odorants.
The partition coefficient between 1-octanol and water was used to indicate the hydrophobicity of each odorant.The odorants listed are those tested in Figure 2 or in Figure 2—figure supplement 2. LogP values were predicted using ACD/LogP software from ACD/Labs. 1-octanol, the most hydrophobic compound tested in Larter et al. (2016), is indicated in red.
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Figure 2—figure supplement 1—source data 1
Source data for logP values plotted in Figure 2—figure supplement 1.
- https://doi.org/10.7554/eLife.51040.006
Figure 2—figure supplement 2
Screen of additional odorants against ORNs in mutant and control basiconic sensilla.
(A–D) An initial screen with additional odorants. In this initial screen at a low n value, only three odorants elicited abnormal responses. * p 0.05, ** p 0.01, non-parametric t test, (SEM; n = 3 ~ 4). (E) Rescreen at higher n value of the three odorants identified in (A–D), and of 5 control odorants. (SEM, n = 10). No significant differences were found in this rescreen.
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Figure 2—figure supplement 2—source data 1
Source data for spike numbers in Figure 2—figure supplement 2.
- https://doi.org/10.7554/eLife.51040.008
Figure 3 with 1 supplement
Responses to stimuli of different temporal dynamics in mutant ab3 sensilla.
(A) Consecutive 500 ms pulses and representative responses to ethyl hexanoate, which activates ab3A, in control and Obp19a-; Obp28a-; Obp83a-,b- flies. (B) Responses to a 5 s pulse. (C) Peri-stimulus time histogram (PSTH) of ab3A responses to consecutive 500 ms pulses of 10−6 ethyl hexanoate delivered at 1 Hz. (D) A PSTH of ab3B responses to a 5 s pulse of 10−6 2-heptanone, n = 4. (E) PSTH of ab3B responses to consecutive 500 ms pulses of 10−6 2-heptanone, n = 5. (F) PSTH of ab3A responses to a 5 s 10−6 ethyl hexanoate pulse, n = 5. The bin sizes for PSTHs is 25 ms. Shaded areas surrounding each curve indicate SEM.
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Figure 3—source data 1
Source data for PSTH numbers in Figure 3.
- https://doi.org/10.7554/eLife.51040.013
Figure 3—figure supplement 1
Responses in ab3 of Obp19a-; Obp28a-; Obp83a-,b- flies to 2-pentanone in two odor stimulus patterns.
(A) PSTH of ab3A responses to consecutive 500 ms pulses of 10−2 2-pentanone delivered at 1 Hz. n = 5. (B) PSTH of ab3A responses to a 5 s pulse of 10−2 2-pentanone, n = 5. Shaded areas surrounding each curve indicate SEM.
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Figure 3—figure supplement 1—source data 1
Source data for PSTH numbers in Figure 3—figure supplement 1.
- https://doi.org/10.7554/eLife.51040.012
Figure 4
Obp19a-; Obp28a- flies have stronger responses to a sample of linoleic acid.
(A) Physiological responses of ab4 sensilla to a sample of linoleic acid are stronger across several concentrations than control responses. * p 0.05, ** p 0.01, non-parametric t test, n = 5. (B) Two-choice oviposition preference paradigm. (C) Shift in oviposition preference for a linoleic acid sample in Obp19a-; Obp28a- mutants.
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Figure 4—source data 1
Source data for oviposition preference index numbers in Figure 4.
- https://doi.org/10.7554/eLife.51040.015
Tables
Key resources table
| Reagents type (species) or resource | Designation | Sourceor reference | Identifiers | Additional information |
|---|---|---|---|---|
| Gene (Drosophila melanogaster) | Obp19a | Flybase | FBgn0031109 | |
| Gene (Drosophila melanogaster) | Obp28a | Flybase | FBgn0011283 | |
| Gene (Drosophila melanogaster) | Obp83a | Flybase | FBgn0011281 | |
| Gene (Drosophila melanogaster) | Obp83b | Flybase | FBgn0010403 | |
| Strain (Drosophila melanogaster) | Canton-S w1118 | Koh et al., 2014 | NA | DOI: 10.1016/j.neuron.2014.07.012 |
| Strain (Drosophila melanogaster) | Obp19a- | This paper | NA | Materials and methods section |
| Strain (Drosophila melanogaster) | Obp28a- | Larter et al., 2016 | NA | DOI: 10.7554/eLife.20242 |
| Strain (Drosophila melanogaster) | Obp83a-,b- | This paper | NA | Materials andmethods section |
| Recombinant DNA reagen | pCFD4 | Addgene | CAT # 49411 | pCFD4-U6:1_U6:3tandemgRNAs |
| Recombinant DNA reagen | pHD-DsRed-attP | Addgene | CAT # 51019 | |
| Chemical compound | Ethyl acetate | MilliporeSigma | CAT # 270989 | CAS # 141-78-6 |
| Chemical compound | 2,3-butanedione | MilliporeSigma | CAT # 11038 | CAS # 431-03-8 |
| Chemical compound | Methyl acetate | MilliporeSigma | CAT # 45999 | CAS # 79-20-9 |
| Chemical compound | Ethyl 3-hydroxybutyrate | MilliporeSigma | CAT # E30603 | CAS # 5405-41-4 |
| Chemical compound | Ethyl hexanoate | MilliporeSigma | CAT # 148962 | CAS # 123-66-0 |
| Chemical compound | 2-heptanone | MilliporeSigma | CAT # 537683 | CAS # 110-43-0 |
| Chemical compound | E2-hexanal | MilliporeSigma | CAT # 132659 | CAS # 6728-26-3 |
| Chemical compound | (±) Geosmin | MilliporeSigma | CAT # G5908 | CAS # 16423-19-1 |
| Chemical compound | Geranyl acetate | MilliporeSigma | CAT # 45896 | CAS # 105-87-3 |
| Chemical compound | Pentyl acetate | MilliporeSigma | CAT # 109584 | CAS # 628-63-7 |
| Chemical compound | 1-octen-3-ol | MilliporeSigma | CAT # O5284 | CAS # 3391-86-4 |
| Chemical compound | Guaiacol | MilliporeSigma | CAT # G5502 | CAS # 90-05-1 |
| Chemical compound | Linoleic acid | MilliporeSigma | CAT # L1376 | CAS # 60-33-3 |
| Chemical compound | Paraffin oil | MilliporeSigma | CAT # 18512 | CAS # 8012-95-1 |
| Chemical compound | Ethanol | MilliporeSigma | CAT # E7023 | CAS # 64-17-5 |
| Chemical compound | Acetone | Fisher Scientific | CAT # 9006–01 | CAS # 67-64-1 |
| Chemical compound | 2-pentanone | MilliporeSigma | CAT # W284203 | CAS # 107-87-9 |
| Chemical compound | 2-butanone | MilliporeSigma | CAT # 34861 | CAS # 78-93-3 |
| Chemical compound | 1-pentanol | MilliporeSigma | CAT # 398268 | CAS # 71-41-0 |
| Chemical compound | Ethyl propionate | MilliporeSigma | CAT # 112305 | CAS # 105-37-3 |
| Chemical compound | Ethyl lactate | MilliporeSigma | CAT # E34102 | CAS # 687-47-8 |
| Chemical compound | Methyl butyrate | MilliporeSigma | CAT # 246093 | CAS # 623-42-7 |
| Chemical compound | Ethyl butyrate | MilliporeSigma | CAT # 75563 | CAS # 105-54-4 |
| Chemical compound | 1-hexanol | MilliporeSigma | CAT # H13303 | CAS # 11-27-3 |
| Chemical compound | E3-hexenol | MilliporeSigma | CAT # 224715 | CAS # 928-97-2 |
| Chemical compound | Hexanal | MilliporeSigma | CAT # 115606 | CAS # 66-25-1 |
| Chemical compound | Hexanoic acid | MilliporeSigma | CAT # 153745 | CAS # 142-62-1 |
| Chemical compound | alpha-terpineol | MilliporeSigma | CAT # 30627 | CAS # 98-55-5 |
| Chemical compound | gamma-hexalactone | MilliporeSigma | CAT # 68554 | CAS # 695-06-7 |
| Chemical compound | 3-methyl-2-buten-1-ol | MilliporeSigma | CAT # 162353 | CAS # 556-82-1 |
| Chemical compound | 6-methyl-5-hepten-2-one | MilliporeSigma | CAT # M48805 | CAS # 110-93-0 |
| Chemical compound | E2-hexenol | MilliporeSigma | CAT # W256230 | CAS # 928-95-0 |
| Chemical compound | Z3-hexenol | MilliporeSigma | CAT # W2564307 | CAS # 928-96-1 |
| Chemical compound | Diethyl succinate | MilliporeSigma | CAT # 07429 | CAS # 123-25-1 |
| Chemical compound | Ethyl trans-2-butenoate | MilliporeSigma | CAT # W348630 | CAS # 623-70-1 |
| Chemical compound | Benzaldehyde | MilliporeSigma | CAT # 418099 | CAS # 100-52-7 |
| Chemical compound | E2-hexenyl acetate | MilliporeSigma | CAT # W256404 | CAS # 2497-18-9 |
| Chemical compound | Citronellyl acetate | MilliporeSigma | CAT # 43646 | CAS # 150-84-5 |
| Chemical compound | Geranyl acetone | MilliporeSigma | CAT # W354201 | CAS # 689-67-8 |
| Chemical compound | o-cresol | MilliporeSigma | CAT # 36922 | CAS # 95-48-7 |
| Chemical compound | Agarose | AmeicanBio | CAT # AB00972 | CAS # 9012-36-6 |
| Chemical compound | Sucrose | MilliporeSigma | CAT # S7903 | CAS # 57-50-1 |
| Software | Prism 7 | GraphPad Prism | RRID:SCR_002798 | |
| Software | AutoSpike32 | Syntech | http://www.ockenfels-syntech.com |
Additional files
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Transparent reporting form
- https://doi.org/10.7554/eLife.51040.016
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Robust olfactory responses in the absence of odorant binding proteins
eLife 8:e51040.
https://doi.org/10.7554/eLife.51040
