Determining how abused drugs affect neurological function requires us to advance our basic understanding of the structure and operation of the synaptic cleft. Traditional tools in neuroscience, which measure averaged properties of bulk samples, have contributed an immense amount of information toward this goal. To understand fully the synapse, however, we must develop new tools of unprecedented sensitivity, tools that are capable of analyzing single synaptic components and observing conformational changes during vesicle fusion at the synapse. The objective of this proposal is to develop such tools and techniques. We plan to use capillary electrophoresis, optical trapping, controlled vesicle fusion, fluorescence resonance energy transfer, biosensor detection, laser induced fluorescence, and single-molecule detection to investigate: (a) the contents of individual synaptic vesicles, and (b) the structure and function of individual proteins involved in the process of vesicle fusion with the cell membrane in the synaptic cleft. In addition to yielding valuable information about the synaptic vesicle cycle and exocytosis, the purpose of this research program is to establish a valuable set of techniques that can be used by the neuroscience community in their investigations of synaptic transmission.

Agency
National Institute of Health (NIH)
Institute
National Institute on Drug Abuse (NIDA)
Type
Research Project (R01)
Project #
5R01DA009873-05
Application #
6164449
Study Section
Special Emphasis Panel (ZRG1-MDCN-1 (01))
Program Officer
Colvis, Christine
Project Start
1996-06-01
Project End
2002-02-28
Budget Start
2000-03-10
Budget End
2001-02-28
Support Year
5
Fiscal Year
2000
Total Cost
$128,977
Indirect Cost
Name
Stanford University
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
009214214
City
Stanford
State
CA
Country
United States
Zip Code
94305
Whelan, Rebecca J; Zare, Richard N (2003) Single-cell immunosensors for protein detection. Biosens Bioelectron 19:331-6
Whelan, Rebecca J; Wohland, Thorsten; Neumann, Lars et al. (2002) Analysis of biomolecular interactions using a miniaturized surface plasmon resonance sensor. Anal Chem 74:4570-6
Wilson, C F; Simpson, G J; Chiu, D T et al. (2001) Nanoengineered structures for holding and manipulating liposomes and cells. Anal Chem 73:787-91
Peleg, G; Ghanouni, P; Kobilka, B K et al. (2001) Single-molecule spectroscopy of the beta(2) adrenergic receptor: observation of conformational substates in a membrane protein. Proc Natl Acad Sci U S A 98:8469-74
Chiu, D T; Wilson, C F; Ryttsen, F et al. (1999) Chemical transformations in individual ultrasmall biomimetic containers. Science 283:1892-5
Lillard, S J; Chiu, D T; Scheller, R H et al. (1998) Separation and characterization of amines from individual atrial gland vesicles of Aplysia californica. Anal Chem 70:3517-24
Chiu, D T; Lillard, S J; Scheller, R H et al. (1998) Probing single secretory vesicles with capillary electrophoresis. Science 279:1190-3
Jardemark, K; Orwar, O; Jacobson, I et al. (1997) Patch clamp detection in capillary electrophoresis. Anal Chem 69:3427-34
Chiu, D T; Hsiao, A; Gaggar, A et al. (1997) Injection of ultrasmall samples and single molecules into tapered capillaries. Anal Chem 69:1801-7
Orwar, O; Jardemark, K; Jacobson, I et al. (1996) Patch-clamp detection of neurotransmitters in capillary electrophoresis. Science 272:1779-82

Showing the most recent 10 out of 12 publications