Protocol detail

Conditional Expression of Alzheimer's Genes in Drosophila

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UAS-Prosβ5 (21794) (70 (link)), UAS-hAPP.Abeta3-42 (64217), and Elav-GS-GAL4;UAS-hAPP;UAS-hBACE1 (56756) stocks were obtained from the Bloomington Drosophila Stock Center (NIH P40OD018537). All lines were maintained on agar-cornmeal-dextrose-yeast growth medium (71 (link)) in a humidified 24°C incubator with 12-hour light/dark cycles. All crosses were set up with female virgins of the respective GAL4 driver line and male UAS-Prosβ5 or W¹¹¹⁸ flies. Progeny were collected within 48 hours of eclosion and allowed to mate on 10% sugar/yeast (SY10) medium (72 (link)) for another 48 hours. Females were then separated and sorted into sets of 25 flies per vial containing SY10 medium supplemented with either 400 μM mifepristone (RU-486) or ethanol vehicle, mixed directly into the food. Blue dye #1 (8 μM) was added to food containing RU-486 for the purpose of identification. Carbon dioxide was used to briefly anesthetize flies for sorting. Flies were moved to vials of fresh medium every 2 to 3 days.

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Chocron E.S., Munkácsy E., Kim H.S., Karpowicz P., Jiang N., Van Skike C.E., DeRosa N., Banh A.Q., Palavicini J.P., Wityk P., Kalinowski L., Galvan V., Osmulski P.A., Jankowska E., Gaczynska M, & Pickering A.M. (2022). Genetic and pharmacologic proteasome augmentation ameliorates Alzheimer’s-like pathology in mouse and fly APP overexpression models. Science Advances, 8(23), eabk2252.

Publication 2022

Elav-GS-GAL4

Agar Carbon dioxide Dextrose Drosophila Ethanol Females Flies Food Male Mifepristone Ru 486 Sugar Yeast

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Variable analysis

independent variables

RU-486 (mifepristone)

dependent variables

Not explicitly mentioned

control variables

Fly stocks and maintenance conditions (agar-cornmeal-dextrose-yeast growth medium, 24°C incubator, 12-hour light/dark cycles)
Crosses setup (female virgins of the respective GAL4 driver line and male UAS-Prosβ5 or W1118 flies)
Progeny collection and mating (within 48 hours of eclosion, 48 hours on 10% sugar/yeast (SY10) medium)
Fly sorting and housing (sets of 25 flies per vial, SY10 medium, moved to fresh medium every 2-3 days)
CO2 anesthesia for fly sorting

positive control

W1118 flies

negative control

Ethanol vehicle

Annotations

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