1996 Fiscal Year Final Research Report Summary
ESTABLISHMENT OF AN ONLINE ASSAY SYSTEM OF INTRACEREBRALLY RELEASED GLUTAMATE BY MEANS OF MICROBIOCENSOR
| Project/Area Number |
06557038
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (A)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 試験 |
|---|
| Research Field |
内科学一般
|
|---|
| Research Institution | University of the Air |
|---|
Principal Investigator |
KITO Shozo University of the Air, Liberal Arts, Professor, 教養学部, 教授 (00010140)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
WATANABE Yasuhiro National Defense Medical College, Professor, 教授 (90127324)
SEMBA Jun'ichi Univ.of the Air, Liberal Arts, Associate Professor, 教養学部, 助教授 (30183429)
|
|---|
| Project Period (FY) |
1994 – 1996
|
| Keywords | glutamate / platinum black sensor / hydrogen peroxide / sensor output / ascorbic acid / platinum tube / glutamate oxidase / glutamine |
|---|
| Research Abstract |
This study was undertaken to assay concentrations of extracellular L-glutamate release from imtracerebral neurons in vivo with a small-volume online sensor made of platinum black consisted of microdialysis probe fixed at the manipulator.
As the first step, in vitro studies to get a linear calibration curve of L-glutamate determined with sensor output with use of the BAS100B/W workstation and low current module to increase the sensitivity and lower the noise level.
The generated hydrogen peroxide is oxidized electrochemically at a Pt electrode held at 600mV and the oxidized current was monitored as a linear relationship between sensor output and glutamate concentration.
As for in vivo experiments, we tried to remove extracellular ascorbic acid which interfere the assay system. We also performed experiments using carbon bulk censer with Os poly meditator instead of platinum black. In this method, the potential of the electrode can be held at 0 mV so that there is no interference by either catecholamine or ascorbic acid.
An another problem in vivo online measurement is that we can not avoid, contamination with glutamine from this assay system, we need a glutamate oxidase which is extremely specific to glutamate. Purification of this kind of enzyme is currently in progress. With this enzyme coming up, establishment of the whole assay system is completed.
|
