一种啮齿类动物脑内微透析记录的清醒活动装置的设计和应用研究  

Design and application of a rodent awake activity device for brain microdialysis recording

作　　者：张钰 周澧 张玲[1,2,3,4] 孙秀萍 李祥磊[1,2,3,4] 秦川 ZHANG Yu;ZHOU Li;ZHANG Ling;SUN Xiuping;LI Xianglei;QIN Chuan(Institute of Laboratory Animal Sciences,Chinese Academy of Medical Sciences(CAMS),Comparative Medicine Center,Peking Union Medical College(PUMC),Beijing 100021,China;National Human Diseases Animal Model Resource Center,Beijing 100021;NHC Key Laboratory of Human Disease Comparative Medicine,Beijing 100021;Beijing Engineering Research Center for Experimental Animal Models of Human Critical Diseases,Beijing 100021)

机构地区：[1]中国医学科学院医学实验动物研究所,北京协和医学院比较医学中心,北京100021 [2]国家人类疾病动物模型资源库,北京100021 [3]国家卫生健康委员会人类疾病比较医学重点实验室,北京100021 [4]北京市人类重大疾病实验动物模型工程技术研究中心,北京100021

出　　处：《中国实验动物学报》2022年第3期350-357,共8页Acta Laboratorium Animalis Scientia Sinica

基　　金：中国科协第五届青年人才托举计划(2019QNRC001);中国医学科学院医学与健康科技创新工程项目(2021-I2M-1-034);国家自然科学基金项目(31970510)。

摘　　要：目的通过自行设计制作啮齿动物脑内微透析记录的清醒活动装置,并进行小鼠清醒活动的脑内微透析采样应用和验证。方法设计并利用相关材料自行制作啮齿动物脑内微透析记录的清醒活动装置,并通过小鼠黑质-纹状体神经环路的多巴胺相关神经递质进行测定和验证。向C57BL/6小鼠的黑质、纹状体埋置微透析探针,手术7 d后进行小鼠清醒活动下的黑质、纹状体的多巴胺(DA)及其代谢中间产物二羟基苯乙酸(DOPAC)两种神经递质的微透析采样。微透析流速为2μL/min,每20 min收集一管样品,共收集200 min,分析两种化学物质浓度随时间的变化。结果(1)啮齿动物脑内微透析记录的清醒活动装置可保证动物在装置内清醒活动的同时可顺利进行微透析采样。(2)啮齿动物脑内样品的可检测到稳定的神经递质含量,样品测得小鼠的多巴胺浓度为黑质(0.11±0.01)nmol/L,纹状体(0.22±0.05)nmol/L;小鼠的DOPAC浓度为黑质(3.38±0.59)nmol/L,纹状体(20.35±6.87)nmol/L。各记录时段之间无明显差异。结论啮齿动物脑内微透析记录的清醒活动装置设计实用、稳定,具有较好的时间、空间应用效果。已申请并授权获批国家专利两项:发明专利(202110525342.5),实用新型专利(202121027247.4)。Objective To design and construct a rodent awake activity device for brain microdialysis recording and achieve the application and verification of brain microdialysis sampling in mice.Methods The rodent awake activity device for brain microdialysis recording was designed and constructed using relevant materials.Dopamine-related neurotransmitters in the substantia nigra-striatum neural circuit in mice were measured and verified by this device.Microdialysis probes were implanted into the substantia nigra and striatum.Recording and microdialysis sampling were performed for 7 days after operation.Dopamine and its metabolic intermediate DOPAC were analyzed in the substantia nigra and striatum.The microdialysis flow rate was 2μL/minute.Samples were collected every 20 minutes for 200 minutes to analyze DA and DOPAC concentration changes over time.Results(1)Mouse brain samples showed stable levels of neurotransmitters.(2)Dopamine concentrations were(0.11±0.01)nmol/L in the substantia nigra and(0.22±0.05)nmol/L in the striatum.DOPAC concentrations were(3.38±0.59)nmol/L in the substantia nigra and(20.35±6.87)nmol/L in the striatum.No significant differences were found between each sampling period.Conclusions The rodent awake activity device for brain microdialysis recording is practical and stable with good applications in time and space dimensions.Two national patents have been applied for and authorized:Patent for invention(202110525342.5)and Patent for utility models(202121027247.4).

关 键 词：神经递质 微透析 清醒活动装置 多巴胺 纹状体 黑质 

分 类 号：Q95-33[生物学—动物学]