This application proposes development of an integrated array of assays to quantitatively measure synaptic function in cultured neurons and acute brain slices, with the potential for scaling up these assays for high- throughput screens. As suggested by RFA-MH-11-40 "Scalable Assays for Unbiased Analysis of Neurobiological Function", this grant does not address a specific biological question, but describes new tools for large-scale analysis of neuronal function. Specifically, our applications proposes in eight specific aims a series of related but independent new assay systems, including new methods of achieving controlled expression of neuronal genes in mice, and new techniques for measuring properties of synaptic function in neurons, ranging from pre- and postsynaptic calcium-signaling over analysis of glutamate receptor trafficking to imaging of neuronal excitation or silencing and various forms of neuronal stress. With these assays, our overall goal is to develop tools to meet the increasingly obvious need for better approaches to study neuro-psychiatric disorders such as autism and schizophrenia. A growing human genetics literature describes many candidate pathogenic genes for these disorders, with a synaptic function likely for some of the implicated genes such as neurexins, suggesting that synapses could represent a pathogenetic hotspot for at least a subset of cases in these diseases. Analyzing candidate disease genes, however, has proven difficult with current approaches that require long-term studies of single genes in time-consuming and expensive experiments. Thus, new approaches that can be scaled up and quantitated without enormous investments in time and effort are needed. The tools we describe here are meant to address this need, at least in part, and are based on a series of technical innovations. The tools can be applied to cultured neurons, acute slices, or in vivo experiments in mice, and primarily use optical detection methods as readout to allow scalability. All of the tools developed under the auspices of this application will be freely and immediately distributed to the community, with the hope that they will become standardized approaches for large-scale interrogation of synaptic function in projects performed throughout the country. This application attempts to address an urgent need for scalable assay systems for analysis of neuronal function, as enunciated by the RFA-MH-11-40. The proposed new assay systems focus on synaptic transmission because neuropharmacology and human genetics identified synaptic transmission as a possible site of impairment in many important brain diseases, including autism and schizophrenia.

Public Health Relevance

This application attempts to address an urgent need for scalable assay systems for analysis of neuronal function, as enunciated by the RFA-MH-11-40. The proposed new assay systems focus on synaptic transmission because neuropharmacology and human genetics identified synaptic transmission as a possible site of impairment in many important brain diseases, including autism and schizophrenia.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS077906-03
Application #
8469589
Study Section
Special Emphasis Panel (ZMH1-ERB-C (05))
Program Officer
Talley, Edmund M
Project Start
2011-07-15
Project End
2015-05-31
Budget Start
2013-06-01
Budget End
2014-05-31
Support Year
3
Fiscal Year
2013
Total Cost
$620,242
Indirect Cost
$212,982
Name
Stanford University
Department
Neurosciences
Type
Schools of Medicine
DUNS #
009214214
City
Stanford
State
CA
Country
United States
Zip Code
94305
Yi, Fei; Danko, Tamas; Botelho, Salome Calado et al. (2016) Autism-associated SHANK3 haploinsufficiency causes Ih channelopathy in human neurons. Science 352:aaf2669
Luo, Fujun; Bacaj, Taulant; Südhof, Thomas C (2015) Synaptotagmin-7 Is Essential for Ca2+-Triggered Delayed Asynchronous Release But Not for Ca2+-Dependent Vesicle Priming in Retinal Ribbon Synapses. J Neurosci 35:11024-33
Yang, Xiaofei; Pei, Jimin; Kaeser-Woo, Yea Jin et al. (2015) Evolutionary conservation of complexins: from choanoflagellates to mice. EMBO Rep 16:1308-17
Acuna, Claudio; Liu, Xinran; Gonzalez, Aneysis et al. (2015) RIM-BPs Mediate Tight Coupling of Action Potentials to Ca(2+)-Triggered Neurotransmitter Release. Neuron 87:1234-47
Burré, Jacqueline; Sharma, Manu; Südhof, Thomas C (2015) Definition of a molecular pathway mediating α-synuclein neurotoxicity. J Neurosci 35:5221-32
Bacaj, Taulant; Ahmad, Mohiuddin; Jurado, Sandra et al. (2015) Synaptic Function of Rab11Fip5: Selective Requirement for Hippocampal Long-Term Depression. J Neurosci 35:7460-74
Bacaj, Taulant; Wu, Dick; Burré, Jacqueline et al. (2015) Synaptotagmin-1 and -7 Are Redundantly Essential for Maintaining the Capacity of the Readily-Releasable Pool of Synaptic Vesicles. PLoS Biol 13:e1002267
Zhang, Bo; Chen, Lulu Y; Liu, Xinran et al. (2015) Neuroligins Sculpt Cerebellar Purkinje-Cell Circuits by Differential Control of Distinct Classes of Synapses. Neuron 87:781-96
Burré, Jacqueline; Sharma, Manu; Südhof, Thomas C (2014) α-Synuclein assembles into higher-order multimers upon membrane binding to promote SNARE complex formation. Proc Natl Acad Sci U S A 111:E4274-83
Tsetsenis, Theodoros; Boucard, Antony A; Araç, Demet et al. (2014) Direct visualization of trans-synaptic neurexin-neuroligin interactions during synapse formation. J Neurosci 34:15083-96

Showing the most recent 10 out of 31 publications