Experimentally induced active and quiet sleep engage non-overlapping transcriptional programs in Drosophila

Sleep in mammals can be broadly classified into two different physiological categories: rapid eye movement (REM) sleep and slow wave sleep (SWS), and accordingly REM and SWS are thought to achieve a different set of functions. The fruit fly Drosophila melanogaster is increasingly being used as a model to understand sleep functions, although it remains unclear if the fly brain also engages in different kinds of sleep as well. Here, we compare two commonly used approaches for studying sleep experimentally in Drosophila: optogenetic activation of sleep-promoting neurons and provision of a sleep-promoting drug, Gaboxadol. We find that these different sleep-induction methods have similar effects on increasing sleep duration, but divergent effects on brain activity. Transcriptomic analysis reveals that drug-induced deep sleep (‘quiet’ sleep) mostly downregulates metabolism genes, whereas optogenetic ‘active’ sleep upregulates a wide range of genes relevant to normal waking functions. This sug..., 2-photon imaging
2-photon imaging was performed as described previously using a ThorLabs Bergamo series 2 multiphoton microscope. Fluorescence was detected with a High Sensitivity GaAsP photomultiplier tube (ThorLabs, PMT2000). GCaMP fluorescence was filtered through the microscope with a 594 dichroic beam splitter and a 525/25nm band pass filter.
For imaging experiments, flies were secured to a custom-built holder. Extracellular fluid (ECF) containing 103 NaCl, 10.5 trehalose, 10 glucose, 26 NaHCO3, 5 C6H15NO6S, 5 MgCl2 (hexa-hydrate), 2 Sucrose, 3 KCl, 1.5 CaCl (dihydrate), and 1 NaH2PO4 (in mM) at room temperature was used to fill a chamber over the head of the fly. The brain was accessed by removing the cuticle of the fly with forceps, and the perineural sheath was removed with a microlance. Flies were allowed to recover from this for one hour before commencement of experiments. Imaging was performed across 18 z-slices, separated by 6µm, with two additional flyback frames. The entir..., , Paper: Experimentally induced active and quiet sleep engage non-overlapping transcriptional programs in Drosophila

Authors:
Niki Anthoney
Lucy A.L. Tainton-Heap
Hang Luong
Eleni Notaras
Amber B. Kewin
Qiongyi Zhao
Trent Perry
Philip Batterham
Paul J. Shaw
Bruno van Swinderen

This README file describes the data package accompanying the above publication.

Files:

2 Photon Data

Files named: 2photon\_FlyX\_23E10Chrimson contain the raw 2photon data and .avi behavioural data for each fly. There are 5 files for each of the 9 flies. These files are affixed with:

\_Experiment

\_ChanB\_Preview

\_Image\_0001\_0001

\_ROIMask

\_ROIs

These files relate to Figure 3C of the above publication.

Files named: 2photon\_flyX\_THIP contain the raw 2photon data and .avi behavioural data for each fly. There are 5 files for each of the 6 flies. These files are affixed with:

\_Experiment

\_ChanB\_Preview

\_Image\_0001\_0001

\_ROIMask

\_ROIs

These files relate to Figure 3D and 4D-4H of the above ...