title: Drosophila glial glutamate transporter Eaat1 is regulated by fringe-mediated notch signaling and is essential for larval locomotion.
abstract: In the mammalian CNS, glial cells expressing excitatory amino acid transporters (EAATs) tightly regulate extracellular glutamate levels to control neurotransmission and protect neurons from excitotoxic damage. Dysregulated EAAT expression is associated with several CNS pathologies in humans, yet mechanisms of EAAT regulation and the importance of glutamate transport for CNS development and function in vivo remain incompletely understood. Drosophila is an advanced genetic model with only a single high-affinity glutamate transporter termed Eaat1. We found that Eaat1 expression in CNS glia is regulated by the glycosyltransferase Fringe, which promotes neuron-to-glia signaling through the Delta-Notch ligand-receptor pair during embryogenesis. We made Eaat1 loss-of-function mutations and found that homozygous larvae could not perform the rhythmic peristaltic contractions required for crawling. We found no evidence for excitotoxic cell death or overt defects in the development of neurons and glia, and the crawling defect could be induced by postembryonic inactivation of Eaat1. Eaat1 fully rescued locomotor activity when expressed in only a limited subpopulation of glial cells situated near potential glutamatergic synapses within the CNS neuropil. Eaat1 mutants had deficits in the frequency, amplitude, and kinetics of synaptic currents in motor neurons whose rhythmic patterns of activity may be regulated by glutamatergic neurotransmission among premotor interneurons; similar results were seen with pharmacological manipulations of glutamate transport. Our findings indicate that Eaat1 expression is promoted by Fringe-mediated neuron-glial communication during development and suggest that Eaat1 plays an essential role in regulating CNS neural circuits that control locomotion in Drosophila.
authors.affiliation: Centre for Research in Neuroscience and Department of Neurology and Neurosurgery, McGill University, Montreal, Quebec H3G 1A4, Canada.
authors.forename: Stephanie M
authors.initials: SM
authors.lastname: Stacey
authors.forename: Nara I
authors.initials: NI
authors.lastname: Muraro
authors.forename: Emilie
authors.initials: E
authors.lastname: Peco
authors.forename: Alain
authors.initials: A
authors.lastname: Labbé
authors.forename: Graham B
authors.initials: GB
authors.lastname: Thomas
authors.forename: Richard A
authors.initials: RA
authors.lastname: Baines
authors.forename: Donald J
authors.initials: DJ
authors.lastname: van Meyel
mesh_terms: D000818:Animals; D004330:Drosophila / Q000502:physiology*; D029721:Drosophila Proteins / Q000235:genetics / Q000502:physiology*; D004594:Electrophysiology; D027341:Excitatory Amino Acid Transporter 1 / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D007814:Larva; D008124:Locomotion / Q000502:physiology*; D009154:Mutation / Q000502:physiology; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics / Q000502:physiology*; D009457:Neuroglia / Q000378:metabolism*; D018408:Patch-Clamp Techniques; D051880:Receptors, Notch / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000502:physiology*; D009435:Synaptic Transmission / Q000502:physiology
chemical_list: D029721:Drosophila Proteins; D027341:Excitatory Amino Acid Transporter 1; D051880:Receptors, Notch; D017351:N-Acetylglucosaminyltransferases; C090576:fng protein, Drosophila