大鼠脊髓突触体的制备及神经递质的定性和定量分析

doi:10.3969/j.issn.1006-9771.2019.06.010

Abstract

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

CLC Number:

R651.2

LIU Zheng, WANG Jing, SHEN Wei, ZHAO Shu-xuan, YUAN Wen-hua, FENG Xiao-fei, ZHOU Hai-yu. Preparation of Rat Spinal Synaptosome and Qualitative and Quantitative Test of Neurotransmitters[J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2019, 25(6): 670-675.

References

1 WhittakerV P, MichaelsonI A, KirklR J. The separation of synaptic vesicles from nerve-ending particles (“synaptosomes”) [J]. Biochem J, 1964, 90(2): 293-303.
2 WhittakerV P. Structure and Function of Synapses [M]. New York: Publishers New York, 1972: 87-88．
3 WilczyńskaK, SimonienkoK, KonarzewskaB, et al. Morphological changes of the brain in mood disorders [J]. Psychiatr Pol, 2018, 52(5): 797-805.
4 NgJ, PapandreouA, HealesS J, et al. Monoamine neurotransmitter disorders: clinical advances and future perspectives [J]. Nat Rev Neurol, 2015, 11(10): 567-584.
5 PerryM, LiQ, KennedyR T. Review of recent advances in analytical techniques for the determination of neurotransmitters [J]. Anal Chim Acta, 2009, 653(1): 1-22.
6 KandimallaR, ReddyP H. Therapeutics of neurotransmitters in Alzheimer's disease [J]. J Alzheimers Dis, 2017, 57(4): 1049-1069.
7 JocaS R, MoreiraF A, WegenerG. Atypical neurotransmitters and the neurobiology of depression [J]. CNS Neurol Disord Drug Targets, 2015, 14(8): 1001-1011.
8 Sanjari MoghaddamH, Zare-ShahabadiA, RahmaniF, et al. Neurotransmission systems in Parkinson's disease [J]. Rev Neurosci, 2017, 28(5): 509-536.
9 SmogorzewskiM J. Skin blood flow and vascular endothelium function in uremia [J]. J Ren Nutr, 2017, 27(6): 465-469.
10 ShahbazA, AzizK, Fransawy AlkomosM, et al. Pheochromocytoma secreting large quantities of both epinephrine and norepinephrine presenting with episodes of hypotension and severe electrolyte imbalance [J]. Cureus, 2018, 10(7): e3050.
11 BeckerC, JickS S, MeierC R. Selective serotonin reuptake inhibitors and cataract risk: a case-control analysis [J]. Ophthalmology, 2017, 124(11): 1635-1639.
12 ŞanlS N, TagueS E, LunteC. Analysis of amino acid neurotransmitters from rat and mouse spinal cords by liquid chromatography with fluorescence detection [J]. J Pharm Biomed Anal, 2015, 107: 217-222.
13 PintoM C, de PaivaM J, Oliveira-LimaO C, et al. Neurochemical study of amino acids in rodent brain structures using an improved gas chromatography-mass spectrometry method [J]. J Chem Neuroanat, 2014, 55: 24-37.
14 MoonJ M, ThapliyalN, HussainK K, et al. Conducting polymer-based electrochemical biosensors for neurotransmitters: a review [J]. Biosens Bioelectron. 2018, 102: 540-552.
15 HayamaT, SakaguchiY, YoshidaH, et al. Binary fluorous alkylation of biogenic primary amines with perfluorinated aldehyde followed by fluorous liquid chromatography-tandem mass spectrometry analysis [J]. Anal Chem, 2012, 84(19): 8407-8414.
16 HylandK. Clinical utility of monoamine neurotransmitter metabolite analysis in cerebrospinal fluid [J]. Clin Chem, 2008, 54(4): 633-641.
17 TenreiroP, RebeloS, MartinsF, et al. Comparison of simple sucrose and percoll based methodologies for synaptosome enrichment [J]. Anal Biochem, 2017, 517: 1-8.
18 ForgacsovaA, GalbaJ, GarrutoR M, et al. A novel liquid chromatography/mass spectrometry method for determination of neurotransmitters in brain tissue: application to human tauopathies [J]. J Chromatogr B Analyt Technol Biomed Life Sci, 2018, 1073: 154-162.
19 ZhangF, MaX L, WangY X, et al. Detection of four different amino acid neurotransmitters in cultured rat neurons and the culture medium by precolumn derivatization high-performance liquid chromatography [J]. Neuroreport, 2016, 27(7): 495-500.
20 PatinF, CrinièreL, FranciaT, et al. Low specificity of urinary 3-methoxytyramine in screening of dopamine-secreting pheochromocytomas and paragangliomas [J]. Clin Biochem, 2016, 49(15): 1205-1208.
21 GalE M, ShermanA D. L-kynurenine: its synthesis and possible regulatory function in brain [J]. Neurochem Res, 1980, 5(3): 223-239.
22 CalhounA, KoJ, GrossbergG T. Emerging chemical therapies targeting 5-hydroxytryptamine in the treatment of Alzheimer's disease [J]. Expert Opin Emerg Drugs, 2017, 22(1): 101-105.
23 SaxenaA R, ChamarthiB, WilliamsG H, et al. Predictors of plasma and urinary catecholamine levels in normotensive and hypertensive men and women [J]. J Hum Hypertens, 2014, 28(5): 292-297.
24 ProkofyevaE, WegenerA, ZrennerE. Cataract prevalence and prevention in Europe: a literature review [J]. Acta Ophthalmol, 2013, 91(5): 395-405.

[1]	LI Fang, HUO Su, DU Jubao, LIU Xiuzhen, LI Xiaoshuang, SONG Weiqun. Effect of transcranial direct current stimulation combined with task-oriented rehabilitation training on forelimb motor dysfunction in rats with spinal cord injury [J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2023, 29(7): 777-781.
[2]	HUANG Zhilin, XU Fashao, SHI Jing, HUANG Gan, LIU Meifang, ZHANG Xiahui. Establishment of rat model of dysphagia after stroke by thread embolism [J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2023, 29(10): 1147-1153.
[3]	QIN Yanqiang, DONG Hao, SUN Yingchun, CHENG Xiankuan, YAO Haijiang. Effects of different acupuncture schemes on neurotransmitters and related inflammatory factors in rats with post-stroke depression [J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2023, 29(1): 30-37.
[4]	MIAO Pei,ZHANG Tong,MI Haixia,ZHANG Weidong. Learning and memory ability and its mechanism in rats with focal cerebral ischemia induced by two filament-occluded methods [J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2022, 28(7): 789-796.
[5]	ZHOU Xiaojue,FENG Jing,PANG Rizhao,LIU Jie,ZHANG Anren. Every-other-day fasting attenuated inflammation in rats after spinal cord injury via the aryl hydrocarbon receptor/suppressor of cytokine signaling 2/nuclear transcription factor-κB signaling pathway [J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2022, 28(5): 544-551.
[6]	SONG Shaofei,HOU Yuanyuan,WANG Yunlei,ZHANG Tong. Effects of circadian misalignment induced by abnormal photoperiod on expression rhythm of clock genes and glucose uptake related genes in gastrocnemius in rats [J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2022, 28(5): 552-558.
[7]	LI Tong,FANG Zhipeng,SHAO Yuping,WANG Ping. Effect of aerobic exercise on learning and memory, and synaptic plasticity of hippocampal neurons for sleep-deprived rats [J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2022, 28(11): 1270-1277.
[8]	Jing-yi WANG,Jie YIN,Jian-cheng LIU,Ri-zhao PANG,Wen-chun WANG. Effect of Iridoid-rich Fraction from Valeriana Jatamansi Jones on Neuron Pyroptosis in Rats with Acute Spinal Cord Injury [J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2021, 27(6): 653-660.
[9]	Wen-mei ZHOU,Tao TAO,Shuang WU,Ting-long WANG,Zheng-yi YANG,Ying ZHANG. Effects of Enriched Environment on Neurological Function and Glucose Metabolism in Ischemic Penumbra in Cerebral Ischemia-reperfusion Injury Rats [J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2021, 27(5): 522-529.
[10]	Qiong-fen WANG,Feng-bo WANG,Ke WANG,Yong-qiang ZHONG,Jiao-jiao WANG. Effect of Electroacupuncture at Fengchi on Astrocytes and Neurons in Rats with Acute Cerebral Infarction [J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2021, 27(3): 302-309.
[11]	Xiang-zhe LI,Jie DING,Qing-hua WANG,Chuan-ming DONG,Tong WANG,Qin-feng WU. Effects of Body Weight-supported Treadmill Training on Neuropathic Pain and Expression of Glutamate Decarboxylase (GAD)-65/67 in Spinal Dorsal Horn of Rats with Spinal Cord Injury [J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2021, 27(2): 131-136.
[12]	Kun AI,Ming XU,Qiong LIU,Shi-feng DENG,Ji-sheng LIU,Fang QI,Xi-qin YI,Qi-rui QU,Hong ZHANG. Effect of Electroacupuncture on Content of Cyclic Adenosine Monophosphate and Protein Kinase A, and Phosphorylation of Myosin Light Chain Kinase in Detrusor of Rats with Detrusor Hyperreflex after Suprasacral Spinal Cord Injury [J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2021, 27(2): 137-144.
[13]	LING Meng-yu,YANG Yi-zhuo,LIU Shuai,YE Chao-qun. Effect of Exercise on Expression of NeuN and SynapsinI in Cortex and Cognitive Function in Rats after Cerebral Ischemia-reperfusion [J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2021, 27(11): 1272-1281.
[14]	PENG Zhi-feng, ZHANG Yi-ping, ZHANG Ji-hong. Effect of Early Exercise Combined with Pentylenetetrazol on Neurologic Function after Cerebral Ischemia-reperfusion in Rats and Its Mechanism [J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2021, 27(1): 27-30.
[15]	CHEN Xiao-ping,WANG Ming-yang,MA Deng-lei,GONG Shi-li,ZHANG Li,LI Ya-li,LI Lin,HU Chao-ying,ZHANG Lan. Effects of Analgecine on Apoptosis and Neuroinflammation in Cerebral Ischemia-reperfusion Injury Rats [J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2020, 26(9): 1038-1044.

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

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Metrics

Comments

Recommended 0