@article{503f944b064e4ab19f4250128d275dfe,
title = "Astrocytic GABA transporter controls sleep by modulating GABAergic signaling in Drosophila circadian neurons",
abstract = "A precise balance between sleep and wakefulness is essential to sustain a good quality of life and optimal brain function. GABA is known to play a key and conserved role in sleep control, and GABAergic tone should, therefore, be tightly controlled in sleep circuits. Here, we examined the role of the astrocytic GABA transporter (GAT) in sleep regulation using Drosophila melanogaster. We found that a hypomorphic gat mutation (gat33-1) increased sleep amount, decreased sleep latency, and increased sleep consolidation at night. Interestingly, sleep defects were suppressed when gat33-1 was combined with a mutation disrupting wide-awake (wake), a gene that regulates the cell-surface levels of the GABAA receptor resistance to dieldrin (RDL) in the wake-promoting large ventral lateral neurons (l-LNvs). Moreover, RNAi knockdown of rdl and its modulators dnlg4 and wake in these circadian neurons also suppressed gat33-1 sleep phenotypes. Brain immunohistochemistry showed that GAT-expressing astrocytes were located near RDL-positive l-LNv cell bodies and dendritic processes. We concluded that astrocytic GAT decreases GABAergic tone and RDL activation in arousal-promoting LNvs, thus determining proper sleep amount and quality in Drosophila.",
keywords = "astrocytes, GABA, GAT, large ventral lateral neurons, sleep",
author = "Ratna Chaturvedi and Tobias Stork and Chunyan Yuan and Marc Freeman and Patrick Emery",
note = "Funding Information: We are grateful to the Vienna Drosophila RNAi Center and the Bloomington Stock Center for providing fly stocks and Prof. K. Koh for transgenic wake miRNA flies. We are thankful to Prof. R. L. Davis, Scripps Research, Florida, for providing anti-RDL antibody and Prof. R. Stanewsky, M{\"u}nster University, Germany, for anti-PER antibody. Anti-PDF, anti-GFP, anti-Pros and, anti-spectrin were obtained from the Developmental Studies Hybridoma Bank, USA. We are grateful to Prof. Mark Wu, Johns Hopkins University, USA, for wake mutant flies; Prof. Junhai Han, Southeast University, Nanjing, China, for the UAS-dnlg4-RNAi fly line; and B. Ganetsky, University of Wisconsin-Madison, USA, and M. Tanouye, University of California, Berkeley, Berkeley, USA, for eas PC80 flies. We also thank Prof. Leslie Griffith and Patricia Goodwin at Brandeis University, USA, for helping us setting up sleep assays in our laboratory as well as Prof. M. Rosbash and Albert Yu for help with building the flybox. We thank all members of the Emery, Weaver and Anaclet lab for insightful discussions on the work and manuscript. This work was supported by a MIRA award from the National Institute of General Medicine Sciences ( 1R35GM118087 ) to P.E. and by a R01 grant from the National Institute of Neurological Disorders and Stroke ( R01 NS053538 ) to M.R.F. Funding Information: We are grateful to the Vienna Drosophila RNAi Center and the Bloomington Stock Center for providing fly stocks and Prof. K. Koh for transgenic wake miRNA flies. We are thankful to Prof. R. L. Davis, Scripps Research, Florida, for providing anti-RDL antibody and Prof. R. Stanewsky, M?nster University, Germany, for anti-PER antibody. Anti-PDF, anti-GFP, anti-Pros and, anti-spectrin were obtained from the Developmental Studies Hybridoma Bank, USA. We are grateful to Prof. Mark Wu, Johns Hopkins University, USA, for wake mutant flies; Prof. Junhai Han, Southeast University, Nanjing, China, for the UAS-dnlg4-RNAi fly line; and B. Ganetsky, University of Wisconsin-Madison, USA, and M. Tanouye, University of California, Berkeley, Berkeley, USA, for easPC80 flies. We also thank Prof. Leslie Griffith and Patricia Goodwin at Brandeis University, USA, for helping us setting up sleep assays in our laboratory as well as Prof. M. Rosbash and Albert Yu for help with building the flybox. We thank all members of the Emery, Weaver and Anaclet lab for insightful discussions on the work and manuscript. This work was supported by a MIRA award from the National Institute of General Medicine Sciences (1R35GM118087) to P.E. and by a R01 grant from the National Institute of Neurological Disorders and Stroke (R01 NS053538) to M.R.F. Conceptualization, R.C. and P.E.; methodology, R.C. T.S. and C.Y.; investigation, R.C. C.Y. and T.S.; writing ? original draft, R.C. and P.E.; writing ? review & editing, T.S. and M.R.F; funding acquisition, M.R.F. and P.E; supervision, P.E. and M.R.F. The authors declare no competing interests. Publisher Copyright: {\textcopyright} 2022 Elsevier Inc.",
year = "2022",
month = may,
day = "9",
doi = "10.1016/j.cub.2022.02.066",
language = "English (US)",
volume = "32",
pages = "1895--1908.e5",
journal = "Current Biology",
issn = "0960-9822",
publisher = "Cell Press",
number = "9",
}