Abstract
In the present protocol, we demonstrate a high-performance liquid chromatography (HPLC) system that enables detection of very low amounts of γ-aminobutyric acid (GABA) (0.03 pmol) and glutamate (0.8 pmol). The HPLC system consists of two pumps, an electrochemical detector, a high-pressure six-way switching valve, a guard column, a microbore column, and a column oven. A microdialysis probe was implanted in the right parietal cortex in rats. Dialysates were collected every 5 min and were split into two equal aliquots for separate analysis of GABA and glutamate. After derivatization with o-phthalaldehyde (OPA), samples were isocratically separated and purified by the guard column. To make the peak of GABA or glutamate appear in an opportune place in a chromatogram, a six-way switching valve was used to control the eluate containing GABA or glutamate to be led to the microbore column and electrochemical detector. By the use of this system, decrease in extracellular concentration of GABA, which precedes the appearance of electrical discharge initiated by hyperbaric oxygen (HBO 2) exposure, was detected by microdialysis at the time resolution of 5 min.
| Original language | English |
|---|---|
| Pages (from-to) | 61-66 |
| Number of pages | 6 |
| Journal | Brain Research Protocols |
| Volume | 14 |
| Issue number | 2 |
| DOIs | |
| Publication status | Published - Feb 2005 |
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Keywords
- Hyperbaric oxygen
- Microdialysis
- Neurotransmitter
- Seizure
ASJC Scopus subject areas
- Neuroscience(all)
Cite this
Measurement of GABA and glutamate in vivo levels with high sensitivity and frequency. / Zhang, Shan; Takeda, Yoshimasa; Hagioka, Shingo; Takata, Ken; Aoe, Hisami; Nakatsuka, Hideki; Yokoyama, Masataka; Morita, Kiyoshi.
In: Brain Research Protocols, Vol. 14, No. 2, 02.2005, p. 61-66.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Measurement of GABA and glutamate in vivo levels with high sensitivity and frequency
AU - Zhang, Shan
AU - Takeda, Yoshimasa
AU - Hagioka, Shingo
AU - Takata, Ken
AU - Aoe, Hisami
AU - Nakatsuka, Hideki
AU - Yokoyama, Masataka
AU - Morita, Kiyoshi
PY - 2005/2
Y1 - 2005/2
N2 - In the present protocol, we demonstrate a high-performance liquid chromatography (HPLC) system that enables detection of very low amounts of γ-aminobutyric acid (GABA) (0.03 pmol) and glutamate (0.8 pmol). The HPLC system consists of two pumps, an electrochemical detector, a high-pressure six-way switching valve, a guard column, a microbore column, and a column oven. A microdialysis probe was implanted in the right parietal cortex in rats. Dialysates were collected every 5 min and were split into two equal aliquots for separate analysis of GABA and glutamate. After derivatization with o-phthalaldehyde (OPA), samples were isocratically separated and purified by the guard column. To make the peak of GABA or glutamate appear in an opportune place in a chromatogram, a six-way switching valve was used to control the eluate containing GABA or glutamate to be led to the microbore column and electrochemical detector. By the use of this system, decrease in extracellular concentration of GABA, which precedes the appearance of electrical discharge initiated by hyperbaric oxygen (HBO 2) exposure, was detected by microdialysis at the time resolution of 5 min.
AB - In the present protocol, we demonstrate a high-performance liquid chromatography (HPLC) system that enables detection of very low amounts of γ-aminobutyric acid (GABA) (0.03 pmol) and glutamate (0.8 pmol). The HPLC system consists of two pumps, an electrochemical detector, a high-pressure six-way switching valve, a guard column, a microbore column, and a column oven. A microdialysis probe was implanted in the right parietal cortex in rats. Dialysates were collected every 5 min and were split into two equal aliquots for separate analysis of GABA and glutamate. After derivatization with o-phthalaldehyde (OPA), samples were isocratically separated and purified by the guard column. To make the peak of GABA or glutamate appear in an opportune place in a chromatogram, a six-way switching valve was used to control the eluate containing GABA or glutamate to be led to the microbore column and electrochemical detector. By the use of this system, decrease in extracellular concentration of GABA, which precedes the appearance of electrical discharge initiated by hyperbaric oxygen (HBO 2) exposure, was detected by microdialysis at the time resolution of 5 min.
KW - Hyperbaric oxygen
KW - Microdialysis
KW - Neurotransmitter
KW - Seizure
UR - http://www.scopus.com/inward/record.url?scp=13844280900&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=13844280900&partnerID=8YFLogxK
U2 - 10.1016/j.brainresprot.2004.03.005
DO - 10.1016/j.brainresprot.2004.03.005
M3 - Article
C2 - 15721811
AN - SCOPUS:13844280900
VL - 14
SP - 61
EP - 66
JO - Brain Research Protocols
JF - Brain Research Protocols
SN - 1385-299X
IS - 2
ER -