This proposal describes a research program in glutamate bioanalysis with ultrasensitivity and simultaneous temporal and spatial resolution. The following aims will be pursued in this proposal: (1) development of glutamate (glu) monitoring techniques for ultrasensitive, ultrafast, and highly selective extracellular glu detection with 0.1 um spatial resolution, (2) development of simultaneous glu and calcium determination methodologies for real-time monitoring, (3) study of glu-calcium hypothesis in ischemic stroke on a subcellular and molecular basis, (4) investigation of multiquantal glu release at single synaptic sites. These goals will be reached with significant bioanalytical development and detailed biological and biomedical studies. The proposed research will capitalize upon the group's recent advancement in nanotechnology for ultrasmall biochemical sensors and in optical detection and imaging at the single molecule level. By the completion of this study, the PI hopes to have gained significant insight into the synaptic release mechanisms and a better understanding of ischemic injury and cell death on a subcellular basis. In addition, tools will be developed to make in-vivo measurements on living samples with a level of resolution heretofore only available in the frontier of physical sciences. The ability to monitor biological events at the subcellular level will not only enable the group to test glu-calcium hypothesis in ischemic stroke and to investigate multiquantal release from single synaptic sites, but also provide new opportunities in a wide range of biomedical studies related to glu and calcium ions.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS039891-02
Application #
6394343
Study Section
Special Emphasis Panel (ZRG1-BMT (01))
Program Officer
Behar, Toby
Project Start
2000-04-10
Project End
2003-03-31
Budget Start
2001-04-01
Budget End
2002-03-31
Support Year
2
Fiscal Year
2001
Total Cost
$214,113
Indirect Cost
Name
University of Florida
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
073130411
City
Gainesville
State
FL
Country
United States
Zip Code
32611
Qhobosheane, Monde; Wu, Donghai; Gu, Yunrong et al. (2004) A two-dimensional imaging biosensor to monitor enhanced brain glutamate release stimulated by nicotine. J Neurosci Methods 135:71-8
Qhobosheane, Monde; Zhang, Peng; Tan, Weihong (2004) Assembly of silica nanoparticles for two-dimensional nanomaterials. J Nanosci Nanotechnol 4:635-40
Zhao, Xiaojun; Tapec-Dytioco, Rovelyn; Wang, Kemin et al. (2003) Collection of trace amounts of DNA/mRNA molecules using genomagnetic nanocapturers. Anal Chem 75:3476-83
Li, Jianwei Jeffery; Tan, Weihong (2003) A real-time assay for DNA sticky-end pairing using molecular beacons. Anal Biochem 312:251-4
Qhobosheane, Monde; Liu, Xiaojing; Gu, Yunrong et al. (2003) Two-dimensional imaging biosensor for the monitoring of lactate released from brain slices. Appl Spectrosc 57:689-96
Li, Jianwei J; Fang, Xiaohong; Tan, Weihong (2002) Molecular aptamer beacons for real-time protein recognition. Biochem Biophys Res Commun 292:31-40
Fang, Xiaohong; Mi, Yanming; Li, Jianwei Jeffery et al. (2002) Molecular beacons: fluorogenic probes for living cell study. Cell Biochem Biophys 37:71-81
Tapec, Rovelyn; Zhao, Xiaojun Julia; Tan, Weihong (2002) Development of organic dye-doped silica nanoparticles for bioanalysis and biosensors. J Nanosci Nanotechnol 2:405-9
Perlette, J; Tan, W (2001) Real-time monitoring of intracellular mRNA hybridization inside single living cells. Anal Chem 73:5544-50
Qhobosheane, M; Santra, S; Zhang, P et al. (2001) Biochemically functionalized silica nanoparticles. Analyst 126:1274-8

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