Information flow in the brain is mediated by transduction of electrical information into chemical information and back again at chemical synapses. Synapses are made up of crucial cellular machineries that orchestrate a balance of membrane traffic to and from the plasma membrane. Our goal is to develop a detailed quantitative understanding of the synapse both in terms of physiological responses to action potential stimuli as well as the molecular underpinnings of its function. We recently developed sensitive approaches that allow us to characterize the heterogeneity of presynaptic function across many nerve terminals from the same cell. These new methods allow us for the first time to determine release probabilities and measures of readily-releasable pools at the single synapse level. The goal of this project is to test hypotheses about the origin of the heterogeneity of these properties. We will test the idea that this heterogeneity arises from synapse to synapse variability in two different molecular control points of neurotransmitter release. In doing so we will also obtain new and rich information about these control points at the single synapse level.
The first aim will examine if the abundance of a Munc-13-1, a critical regulator of exocytosis, accounts for the heterogeneity. To do this we will examine the biophysical parameters of release at synapses in neurons derived from Munc-13-1-ECFP knockin mice. Calibration procedures will allow us to determine the absolute number of these regulatory molecules at each nerve terminal which will then be compared to functional readouts at the same terminal. We will additionally use shRNA-based manipulation of this protein will allow us to determine the molecular dose-response relationship for function at a very detailed level.
Our second Aim will use this same mapping approach of presynaptic properties and determine how function is correlated with the specific types of calcium channels present at each synapse and how the abundance of functional channels influences key parameters of neurotransmitter release at the single synapse level. Finally we will examine how G-protein based modulation of synapses varies from across a population of synapses and how it impacts function.

Public Health Relevance

Information flow in the brain is mediated by transduction of electrical information into chemical information and back again at chemical synapses. The functioning of the human brain relies on the careful orchestration of delivering neurotransmitter-laden vesicles to sites at nerve terminals where they can be used to deliver this chemical message on demand. Many known genetic mutations in diseases such as Parkinson's disease, migraine headache and schizophrenia are linked to proteins that control synapse function. Our work is aimed at understanding the machinery at a molecular level to better ensure the success of future therapies for these types of neuronal diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of Mental Health (NIMH)
Type
Research Project (R01)
Project #
5R01MH085783-04
Application #
8389677
Study Section
Neurotransporters, Receptors, and Calcium Signaling Study Section (NTRC)
Program Officer
Asanuma, Chiiko
Project Start
2009-12-02
Project End
2014-11-30
Budget Start
2012-12-01
Budget End
2013-11-30
Support Year
4
Fiscal Year
2013
Total Cost
$401,544
Indirect Cost
$163,944
Name
Weill Medical College of Cornell University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
060217502
City
New York
State
NY
Country
United States
Zip Code
10065
Hoppa, Michael B; Gouzer, Geraldine; Armbruster, Moritz et al. (2014) Control and plasticity of the presynaptic action potential waveform at small CNS nerve terminals. Neuron 84:778-89
Kim, Sung Hyun; Ryan, Timothy A (2013) Balance of calcineurin A? and CDK5 activities sets release probability at nerve terminals. J Neurosci 33:8937-50
Pan, Ping-Yue; Ryan, Timothy A (2012) Calbindin controls release probability in ventral tegmental area dopamine neurons. Nat Neurosci 15:813-5
Ariel, Pablo; Ryan, Timothy A (2012) New insights into molecular players involved in neurotransmitter release. Physiology (Bethesda) 27:15-24
Ariel, Pablo; Ryan, Timothy A (2010) Optical mapping of release properties in synapses. Front Neural Circuits 4:
Kim, Sung Hyun; Ryan, Timothy A (2010) CDK5 serves as a major control point in neurotransmitter release. Neuron 67:797-809