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Glutamate release from isolated synaptoneurosomes from the mouse cortex
JIN Guo-rong DING Cai-yun WANG Yi-min LIU Jian-rong WEI Hong-en*
Acta Anatomica Sinica ›› 2014, Vol. 45 ›› Issue (6) : 861-864.
Glutamate release from isolated synaptoneurosomes from the mouse cortex
Objective To detect the release of glutamate from the isolated synaptoneurosomes from the mouse cortex. Methods The mouse cortex was dissected, homogenized and filtered through three layers of 147 μm nylon meshes. The synaptoneurosomes were resuspended after centrifuged. An enzyme-linked fluorescence detection method was used to detect the glutamate release, which utilized glutamate dehydrogenase, and the reduction of nicotinamide adenine dinucleotide phosphate(NADP) to NADPH. The glutamate standard curve was set up and then the glutamate release from isolated synaptoneurosomes was tested and analyzed in three independent experiments. Results A good linear relationship was found between the glutamate concentrations and fluorescence values. The release of glutamate from synaptoneurosomes significantly increased after depolarization agent KCl added and gradually reached a plateau. Glutamate release was increased more when Triton X-100 was added. The inactive synaptoneurosomes after one freeze-thaw cycle had a different curve of glutamate release. Conclusion The release of excitatory neurotransmitter glutamate from synaptoneurosomes of the mouse cortex isolated using the improved method is well studied.
Synaptoneurosome / Glutamate / Release / Enzymatic reaction / Mouse
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