This proposal focuses on two types of change in synapse morphology: expansion of postsynaptic membranes and formation of presynaptic boutons. Anatomical specializations are a hallmark of synapses. On the postsynaptic side, anatomical specializations include dendritic spines and membrane folds. On the presynaptic side, an enlargement of the neurite to form a rounded presynaptic bouton or en passant swelling is a nearly universal feature of synapses.
Aim 1 of this proposal tests a specific hypothesis for how activity can regulate anatomical changes at the postsynapse in the fly neuromuscular junction and in mammalian dendritic spines. In particular, it seeks to elucidate the role of Ral as a mediator of activity-dependent anatomical plasticity. We have uncovered a novel pathway for synaptic plasticity in which the small GTPase Ral, by activating the exocyst complex, serves a central role in transducing Ca2+ influx from glutamate receptors into enhanced transport of membrane to the postsynaptic region. In consequence, the membrane area of the postsynaptic specialization at the fly neuromuscular junction (NMJ) expands in an activity-dependent manner. This proposal asks what patterns of synaptic activity are necessary to recruit the exocyst and how Ral comes to be localized to postsynaptic membranes. It goes on to investigate the significance of Ral for the formation of dendritic spines in the mammalian CNS.
Aim 2 uses Drosophila as a model system in which to uncover the machinery that allows synaptic boutons to form. The maturation of presynaptic terminals from growth cones to synaptic boutons is a critical late step in synaptogenesis but poorly understood. Neither the cytoskeletal elements that underlie the shape change nor signaling molecules that trigger it are known. In a previous mutant screen in Drosophila we discovered that mutations in 2-3 arrest synaptogenesis after initial synapse formation and prevent the formation of synaptic boutons at the fly NMJ. We propose to pursue the process of bouton formation in greater detail by a combination of biochemical and genetic approaches to uncover additional players in the formation of this poorly understood aspect of synaptic anatomy. Therefore, in Aim 2A we will use biochemical methods to identify binding partners for 2-3 and determine if they are required for bouton formation.
In Aim 2 B we will uncover additional players in bouton formation through a mutant screen of the Drosophila genome at the embryonic NMJ.

Public Health Relevance

Synapse formation and synaptic plasticity are at the core of how the brain wires and are critical to issues as wide ranging as disorders of intellectual development and the regulation of how much we eat. This proposal examines the mechanisms by which synapses acquire their mature form and grow in response to the activity of a circuit or an environmental change such as a fasting diet.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
2R01NS041062-14
Application #
8504104
Study Section
Synapses, Cytoskeleton and Trafficking Study Section (SYN)
Program Officer
Talley, Edmund M
Project Start
2000-04-01
Project End
2018-04-30
Budget Start
2013-05-01
Budget End
2014-04-30
Support Year
14
Fiscal Year
2013
Total Cost
$558,400
Indirect Cost
$240,374
Name
Children's Hospital Boston
Department
Type
DUNS #
076593722
City
Boston
State
MA
Country
United States
Zip Code
02115
Teodoro, Rita O; Pekkurnaz, Gulcin; Nasser, Abdullah et al. (2013) Ral mediates activity-dependent growth of postsynaptic membranes via recruitment of the exocyst. EMBO J 32:2039-55
Murthy, Mala; Teodoro, Rita O; Miller, Tamara P et al. (2010) Sec5, a member of the exocyst complex, mediates Drosophila embryo cellularization. Development 137:2773-83
Higashi-Kovtun, Misao E; Mosca, Timothy J; Dickman, Dion K et al. (2010) Importin-beta11 regulates synaptic phosphorylated mothers against decapentaplegic, and thereby influences synaptic development and function at the Drosophila neuromuscular junction. J Neurosci 30:5253-68
Mosca, Timothy J; Schwarz, Thomas L (2010) The nuclear import of Frizzled2-C by Importins-beta11 and alpha2 promotes postsynaptic development. Nat Neurosci 13:935-43
Mosca, Timothy J; Schwarz, Thomas L (2010) Drosophila Importin-?2 is involved in synapse, axon and muscle development. PLoS One 5:e15223
Kurshan, Peri T; Oztan, Asli; Schwarz, Thomas L (2009) Presynaptic alpha2delta-3 is required for synaptic morphogenesis independent of its Ca2+-channel functions. Nat Neurosci 12:1415-23
Dickman, Dion K; Kurshan, Peri T; Schwarz, Thomas L (2008) Mutations in a Drosophila alpha2delta voltage-gated calcium channel subunit reveal a crucial synaptic function. J Neurosci 28:31-8
Miech, Claudia; Pauer, Hans-Ulrich; He, Xi et al. (2008) Presynaptic local signaling by a canonical wingless pathway regulates development of the Drosophila neuromuscular junction. J Neurosci 28:10875-84
Khodosh, Rita; Augsburger, Adela; Schwarz, Thomas L et al. (2006) Bchs, a BEACH domain protein, antagonizes Rab11 in synapse morphogenesis and other developmental events. Development 133:4655-65
Johnson, Karl G; Tenney, Alan P; Ghose, Aurnab et al. (2006) The HSPGs Syndecan and Dallylike bind the receptor phosphatase LAR and exert distinct effects on synaptic development. Neuron 49:517-31

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