title: The homeobox transcription factor Even-skipped regulates acquisition of electrical properties in Drosophila neurons.
abstract: BACKGROUND
  While developmental processes such as axon pathfinding and synapse formation have been characterized in detail, comparatively less is known of the intrinsic developmental mechanisms that regulate transcription of ion channel genes in embryonic neurons. Early decisions, including motoneuron axon targeting, are orchestrated by a cohort of transcription factors that act together in a combinatorial manner. These transcription factors include Even-skipped (Eve), islet and Lim3. The perdurance of these factors in late embryonic neurons is, however, indicative that they might also regulate additional aspects of neuron development, including the acquisition of electrical properties.
  
  
  RESULTS
  To test the hypothesis that a combinatorial code transcription factor is also able to influence the acquisition of electrical properties in embryonic neurons we utilized the molecular genetics of Drosophila to manipulate the expression of Eve in identified motoneurons. We show that increasing expression of this transcription factor, in two Eve-positive motoneurons (aCC and RP2), is indeed sufficient to affect the electrical properties of these neurons in early first instar larvae. Specifically, we observed a decrease in both the fast K+ conductance (IKfast) and amplitude of quantal cholinergic synaptic input. We used charybdotoxin to pharmacologically separate the individual components of IKfast to show that increased Eve specifically down regulates the Slowpoke (a BK Ca2+-gated potassium channel), but not Shal, component of this current. Identification of target genes for Eve, using DNA adenine methyltransferase identification, revealed strong binding sites in slowpoke and nAcRalpha-96Aa (a nicotinic acetylcholine receptor subunit). Verification using real-time PCR shows that pan-neuronal expression of eve is sufficient to repress transcripts for both slo and nAcRalpha-96Aa.
  
  
  CONCLUSIONS
  Taken together, our findings demonstrate, for the first time, that Eve is sufficient to regulate both voltage- and ligand-gated currents in motoneurons, extending its known repertoire of action beyond its already characterized role in axon guidance. Our data are also consistent with a common developmental program that utilizes a defined set of transcription factors to determine both morphological and functional neuronal properties.
authors.affiliation: Neuroscience Group, Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, UK. Edward.Pym@gmail.com
authors.forename: Edward C G
authors.initials: EC
authors.lastname: Pym
authors.forename: Tony D
authors.initials: TD
authors.lastname: Southall
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Mee
authors.forename: Andrea H
authors.initials: AH
authors.lastname: Brand
authors.forename: Richard A
authors.initials: RA
authors.lastname: Baines
mesh_terms: D000109:Acetylcholine / Q000494:pharmacology; D000818:Animals; D030801:Animals, Genetically Modified; D018999:Charybdotoxin / Q000494:pharmacology; D004307:Dose-Response Relationship, Radiation; D004330:Drosophila; D029721:Drosophila Proteins / Q000235:genetics* / Q000378:metabolism*; D004553:Electric Conductivity; D004558:Electric Stimulation / Q000379:methods; D004625:Embryo, Nonmammalian; D005786:Gene Expression Regulation / Q000235:genetics; D018398:Homeodomain Proteins / Q000235:genetics* / Q000378:metabolism* / Q000502:physiology; D051036:Large-Conductance Calcium-Activated Potassium Channels / Q000235:genetics / Q000378:metabolism; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology* / Q000528:radiation effects; D009046:Motor Neurons / Q000187:drug effects / Q000502:physiology* / Q000528:radiation effects; D009498:Neurotoxins / Q000494:pharmacology; D018408:Patch-Clamp Techniques / Q000379:methods; D011188:Potassium / Q000494:pharmacology; D011485:Protein Binding / Q000187:drug effects / Q000235:genetics; D011978:Receptors, Nicotinic / Q000235:genetics / Q000378:metabolism; D013569:Synapses / Q000187:drug effects / Q000502:physiology / Q000528:radiation effects; D013696:Temperature; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism*
chemical_list: D029721:Drosophila Proteins; D018398:Homeodomain Proteins; D051036:Large-Conductance Calcium-Activated Potassium Channels; D009498:Neurotoxins; D011978:Receptors, Nicotinic; D014157:Transcription Factors; C052837:eve protein, Drosophila; C494844:slo protein, Drosophila; D018999:Charybdotoxin; D000109:Acetylcholine; D011188:Potassium