Determination Of Amino Acid Neurotransmitter Mediators In Rat Spinal Cord Synaptosomes

Objectives: To qualitatively and quantitatively detect the amino acid transmitters under different treatment conditions of synaptosomes,and to study the physiological activity of rat spinal cord synaptosomes obtained by Percoll density gradient centrifugation.Synaptic body morphology was observed by transmission electron microscopy to verify the integrity of the synaptosome.Methods: The prepared rat spinal synaptosomes were mixed and divided into three groups:normal incubation group,potassium chloride stimulation group,ultrasonic cell disruption fragmentation group;transmission electron microscope was used to observe the morphological changes of three groups of spinal cord synaptosomes under different treatment;use of high performance liquid chromatography to detect the type and content of amino acids in the suspension supernatant of spinal synaptosomes in normal incubation group,potassium chloride stimulation group,and ultrasonic cell disruptor group.Results:The morphology of synaptosome is complete,and the morphology of synaptosome in each treatment group is different.The neurotransmitters contained in rat spinal synaptosomes were aspartic acid,glycine,glutamic acid,and r-aminobutyric acid,although taurine was detected in the spinal homogenate supernatant,no taurine was detected in synaptosomes;aspartate and glutamate released significantly increased(P <0.01)in the potassium chloride stimulation group,with statistical significance,glycine and r-aminobutyric acid showed an upward trend,but no statistically significant.In addition,we detected asparagine in rat spinal synaptosomes with statistical significance between the two groups;we used ultrasonic cell disruption system to disrupt synaptosomes and detect amino acid transmitters at the same protein concentration significantly lower than the other two groups,this phenomenon needs further study.Conclusions: The neurotransmitters contained in rat spinal synaptosomes were aspartic acid,glycine,glutamic acid,and r-aminobutyric acid and rat spinal synaptosomes have physiological activity.