Human disorders caused by neuronal ion channel dysfunction are a major source of pain, suffering, and economic hardship. Their amelioration can be greatly facilitated by understanding the normal in vivo physiological functions served by the large number of biophysically distinct ion channels present in all animals. Our long-term goal is the development of a generally-applicable transgenic toolkit that will permit the testing of hypotheses concerning the physiological functions of particular ion channel subtypes in specific neural circuits in intact behaving animals. Our approach is based on the """"""""tethered toxin"""""""" technology, wherein peptide ion channel blockers from venomous predators are expressed as fusion proteins tethered to the extracellular side of the plasma membrane. Preliminary studies applying this approach indicate that tethered spider toxins function as cell-autonomous ion channel blockers with their expected target selectivity when expressed in specific neuronal circuits in the brains of transgenic Drosophila melanogaster fruit flies. Preliminary studies also indicate that the venoms of Australian funnel-web spiders of the Atracinae family contain a vast diversity of uncharacterized peptide toxins expected to target a wide variety of ion channel subtypes. The proposed aims are thus directed at (1) identifying novel spider toxins with high potency against neuronal ion channels and (2) determining the molecular identities of the ion channel targets of each identified toxin.
The first aim will be achieved by screening for specific behavioral effects of expressing numerous different tethered funnel-web spider toxins in behavioral control circuits in the brains of transgenic flies.
The second aim will be achieved by using a combination of in vitro and in vivo electrophysiological approaches to identify the molecular target(s) of each toxin identified in the first aim. The proposed research will enable the entire Drosophila neurobiology community to begin to test hypotheses concerning the roles of particular ion channel subtypes in specific neural circuits in intact behaving animals that have been refractory to traditional approaches. Because of the extensive conservation of biophysical and physiological mechanisms of neuronal function between flies and mammals, the novel toxins we identify will not only be of tremendous use for in vivo fly neurobiology, but also as pharmacological reagents for probing the structure and function of mammalian ion channels in health and disease. ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS056443-02
Application #
7271132
Study Section
Special Emphasis Panel (ZRG1-MDCN-K (90))
Program Officer
Silberberg, Shai D
Project Start
2006-08-02
Project End
2011-01-31
Budget Start
2007-02-01
Budget End
2008-01-31
Support Year
2
Fiscal Year
2007
Total Cost
$368,980
Indirect Cost
Name
Yale University
Department
Physiology
Type
Schools of Medicine
DUNS #
043207562
City
New Haven
State
CT
Country
United States
Zip Code
06520
Chen, Dandan; Sitaraman, Divya; Chen, Nan et al. (2017) Genetic and neuronal mechanisms governing the sex-specific interaction between sleep and sexual behaviors in Drosophila. Nat Commun 8:154
Ghosh, D Dipon; Sanders, Tom; Hong, Soonwook et al. (2016) Neural Architecture of Hunger-Dependent Multisensory Decision Making in C. elegans. Neuron 92:1049-1062
Raccuglia, Davide; McCurdy, Li Yan; Demir, Mahmut et al. (2016) Presynaptic GABA Receptors Mediate Temporal Contrast Enhancement in Drosophila Olfactory Sensory Neurons and Modulate Odor-Driven Behavioral Kinetics. eNeuro 3:
Sitaraman, Divya; Aso, Yoshinori; Rubin, Gerald M et al. (2015) Control of Sleep by Dopaminergic Inputs to the Drosophila Mushroom Body. Front Neural Circuits 9:73
Sitaraman, Divya; Aso, Yoshinori; Jin, Xin et al. (2015) Propagation of Homeostatic Sleep Signals by Segregated Synaptic Microcircuits of the Drosophila Mushroom Body. Curr Biol 25:2915-27
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Aso, Yoshinori; Sitaraman, Divya; Ichinose, Toshiharu et al. (2014) Mushroom body output neurons encode valence and guide memory-based action selection in Drosophila. Elife 4:e04580
Choi, Ben Jiwon; Imlach, Wendy L; Jiao, Wei et al. (2014) Miniature neurotransmission regulates Drosophila synaptic structural maturation. Neuron 82:618-34
Aso, Yoshinori; Sitaraman, Divya; Ichinose, Toshiharu et al. (2014) Mushroom body output neurons encode valence and guide memory-based action selection in Drosophila. Elife 3:e04580
Kunst, Michael; Hughes, Michael E; Raccuglia, Davide et al. (2014) Calcitonin gene-related peptide neurons mediate sleep-specific circadian output in Drosophila. Curr Biol 24:2652-64

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