Drosophila Courtship Conditioning: En metode til at teste læring og hukommelse i fluer
Published: April 30, 2023
Abstract
Kilde: Koemans, TS, et al. Drosophila Courtship Conditioning Som et mål for læring og hukommelse. J. Vis. Exp. (2017).
Denne video beskriver en klassisk conditioning assay, der tester læring og hukommelse i Drosophila, kaldet frieri conditioning. Analysen er baseret på reduktion af mandlige frieri efter at have oplevet en afvisning af en ikke-modtagelig premated kvinde. Eksempelprotokollen viser en opsætning af den procedure, der kan bruges til at vurdere kort- og langtidshukommelse.
Protocol
Denne protokol er et uddrag fra Koemans et al., Drosophila Courtship Conditioning As a Measure of Learning and Memory, J. Vis. Exp. (2017). BEMÆRK: I den protokol, der er beskrevet nedenfor, beskrives en replikat af indsamling, træning og test. For at teste reproducerbarheden af resultaterne skal disse trin gentages parallelt, på flere dage og med separate grupper af fluer (tabel 1). Protokollen er baseret på en 10 d…
Representative Results
Figur 1: Fastlæggelse af frieriindekset og eksperimentel oversigt. (A)Billeder, der viser stereotype mandlige frieri adfærd over for en kvindelig flyve. Forskellige stadier af frieri adfærd er vist: orientering (I), følgende (II), vingevibrationer (udvidelse) og trykke (III), slikke (IV), og forsøg på samleje (V). (B) Skematisk oversigt over trænings- …
Materials
| P{KK101437}VIE-260B | VDRC | 101437 | Dhat-at-RNAi in 60100 background |
| P{KK108109}VIE-260B | – | – | Control-RNAi in 60100 background (gift from K. Keleman) |
| w+, UAS-dcr2/yhh;;elav-Gal4 (III) | – | – | panneuronal driver line |
| Containers for plant tissue culture | VWR | 960177 | 175 mL plastic vials |
| Folded filters | Whatman | 10311643 | Filter paper to enlarge area flies can pupate on |
| Flat-bottom blocks (96-wells) | Qiagen | 19579 | Used for housing blocks |
| MicroAmp Clear Adhesive Film | Applied Biosystems | 4306311 | PCR adhesive film as lid on flat-bottom blocks |
| Razor blade | – | – | Any sharp will do |
| Needle | – | – | 0.8 mm diameter |
| Aspirator | – | – | Cut a 1mL pipet tip with scissors in order to have two pieces. The narrow tip of the pipettip is placed as fly entrance in a ~80 cm flexible hose. To prevent a fly from getting in the hose, a normal piece of cotton or small mesh gaze is placed in between the tip and the hose. The other half of the pipettip can be used as mouth piece at the end of the hose. |
| Courtship chambers | – | – | file S1 can be opened with indicated CAD software |
| Camcorder | Sony | – | camera specification: >4M pixels, full HD. |
| For manual quantification, any simple video recording device has the potential to produce a video of sufficient quality to observe courtship behavior accurately. For automated quantification, there will likely be different requirements depending on the software to be used, and users should investigate this thoroughly if automated quantification is desired. | |||
| Name | Company | Catalog Number | Comments |
| power food | |||
| Agar | Sigma | A7002 | |
| Yeast | Bruggeman | – | |
| Yeast extract | MP biomedicals | 0210330391 | |
| Peptone | Sigma | P6838 | |
| Sucrose | Sigma | S9378 | |
| Glucose | Sigma | G7021 | |
| MgSO4 | Sigma | M2643 | |
| CaCl2 | Merck | 1023780500 | |
| Methylparabene (CAUTION) | Sigma | H5501 | |
| Propionic acid (CAUTION) | Sigma | P1386 | |
| Demineralized water | – | ||
| Yeast paste | – | yeast grains and water mixture in a 1:1 ratio | |
| Name | Company | Catalog Number | Comments |
| normal food | |||
| Agar | MP biomedicals | 215017890 | |
| Yeast | bruggeman | – | |
| Corn flour | de Molen | – | |
| Sugar | de Molen | – | |
| Methylparabene (CAUTION) | Sigma | H5501 | |
| Propionic acid (CAUTION) | Sigma | P1386 | |
| Demineralized water | – |
Cite This Article
Drosophila Courtship Conditioning: A Method to Test Learning and Memory In Flies. J. Vis. Exp. (Pending Publication), e20068, doi: (2023).