《Chinese Journal of Rehabilitation Theory and Practice》 ›› 2019, Vol. 25 ›› Issue (6): 670-675.doi: 10.3969/j.issn.1006-9771.2019.06.010

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Preparation of Rat Spinal Synaptosome and Qualitative and Quantitative Test of Neurotransmitters

LIU Zheng1,2, WANG Jing3, SHEN Wei1,2, ZHAO Shu-xuan1,2, YUAN Wen-hua1,2, FENG Xiao-fei1,2, ZHOU Hai-yu1   

  1. 1.Department of Orthopedics, Second Hospital of Lanzhou University, Lanzhou, Gansu 730030, China;
    2.Key Laboratory of Bone and Joint Diseases of Gansu Province, Lanzhou, Gansu 730030, China;
    3.Department of Special Medicine, The 943th Hospital of the Chinese People's Liberation Army, Wuwei, Gansu 733000, China
  • Received:2019-01-02 Revised:2019-02-01 Online:2019-06-25
  • Contact: ZHOU Hai-yu. E-mail: gslzzhy2004@163.com
  • Supported by:
    Natural Science Foundation of Gansu Province (No. 17JR5RA189)

Abstract: Objective To prepare rat spinal synaptosomes, and to conduct qualitative and quantitative analysis of various neurotransmitters in rat spinal synaptosomes to explore the physiological activities.Methods Rat spinal synaptosomes were prepared by discontinuous Percoll density gradient centrifugation and their morphology and structure were observed under transmission electron microscopy. The properties and content of neurotransmitters in rat spinal synaptosomes were determined by ultra performance liquid chromatography-tandem mass spectrometry.Results Under the experimental conditions, rat spinal synaptosomes were found to contain dopamine, norepinephrine, serotonin, homovanillic acid, 3,4-dihydroxybenzene, 5-hydroxyindoleacetic acid, methoxytyramine, tryptophan, γ-aminobutyric acid, kynurenine, 3-hydroxy kynurenine and 3-hydroxy-2-aminobenzoic acid, and so on. The corresponding neurotransmitter contents were detected, and the corresponding linear relationship and correlation coefficient were obtained.Conclusion Discontinuous Percoll density gradient centrifugation could prepare rat spinal synaptosomes and detect neurotransmitters and their contents in rat spinal synaptosomes.

Key words: spinal synaptosomes, neurotransmitters, ultra performance liquid chromatography, mass spectrometer, rats

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