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Title: 大鼠纹状体多巴胺分泌的电化学记录技术及其在体(in vivo)研究
Author: 姚伟
Degree Level: 博士
Issued Date: 2007-01-09
Degree Grantor: 中国科学院上海生命科学研究院
Place of Degree Grantor: 上海生命科学研究院
Supervisor: 周专
Keyword: 动作电位 ; 编码 ; 多巴胺 ; 纹状体
Alternative Title: Electrochemica technique of dopamine release and study in vivo
Major: 神经生物学
Abstract: 中枢神经系统(CNS)中信息交流依赖于突触传递,其主要由突触前分泌的神经递质所介导,它决定了大脑能否正常行使功能。实时监测神经递质在脑内的释放情况,对于了解神经递质分泌异常所导致的脑功能异常形成机制至关重要。在体 (in vivo) 微碳纤电极(CFE)电化学监测技术可以用于探测脑内易氧化类物质,其中包括去甲肾上腺素,肾上腺素和多巴胺(DA)三种儿茶酚胺类神经递质。DA是CNS中一种重要的神经递质,其主要由胞体分布在中脑黑质致密部和腹侧背盖区,轴突末梢主要分布在纹状体,伏隔核,海马,前额叶皮层等区域的DA能神经元末梢分泌。DA在运动,学习记忆,情绪,奖赏等正常生理功能中发挥着重要作用。DA分泌的异常是多种神经精神类疾病发生发展的一个重要病因,例如,帕金森病,药物成瘾,精神分裂症,抑郁症,亨廷顿舞蹈病等。在本研究中,为了获得更高效,更敏感的CFE探测脑内DA的分泌,我们改进了以往用导电胶制作的CFE (C_CFE),采用已二醇已醚(EGME)作为导电介质。与前者相比,用EGME制作的CFE(Y_CFE)所需制作材料少,制作方便,生物学性质稳定,探测DA敏感度高,可用于in vivo生物学实验。在我们以往的研究中,用4个参数n、m、f、d定义了动作电位(AP)复杂的发放模式F(n,m,f,d),其中设一串AP为一个“Burst”,在单个Burst内的AP数目为“n”,频率为“f”,在一次刺激中的Burst数目为“m”,两个相邻Burst之间的时间间隔为“d”。我们发现,除了AP频率外,对于一组给定的AP数,另一个AP编码的参数,AP的分组数(m),能有效调节大鼠肾上腺嗜铬细胞分泌,既“m”作用。然而,在体中“m”作用是否能有效地调节神经递质的分泌还不清楚。 我们用不同的AP编码模式电刺激麻醉状态下大鼠DA能神经元投射纤维前脑内侧束,并用Y_CFE在纤维末梢的投射区纹状体,实时记录了DA的分泌。另外,我们在纹状体脑片,in vivo伏隔核,海马和前额叶皮层观察了DA的分泌。 我们的工作表明:与C_CFE相比,Y_CFE制作所需材料少,制作方便,生物学性质稳定,探测DA敏感度高,可用于in vivo生物学实验。Y_CFE对DA测量的结果表明,在in vivo 情况下,“m”作用能效地调节DA分泌,而且随编码参数的不同,“m”作用也发生不同的变化。
English Abstract: Information communication in central nervous system (CNS) depends on synaptic transmission mainly mediated by presynaptic neurotransmitter release, which plays a crucial role in normal brain function. Real-time measurement of neurotransmitter release in brain is very important to study the mechanisms in which abnormality of neurotrnsmitter release leads to that of brain function. Carbon fiber microelectrode (CFE ) with electrochemical technique in vivo is employed to measure easily oxidized substances including noradrenaline, adrenaline and dopamine (DA) belonging to catecholamine. DA is an important neurotransmitter in CNS, which is released by DAnergic terminals distributing in striatum, nucleus accumbens, hippocampus and medial prefrontal cortex etc. DA cell bodys locate in substantia nigra and ventral tegmental area. DA is closely related with many normally physiological functions including movement, learning and memory, mood and reward. The abnormality of DA release is a critical cause of many kinds of nerve and psychiatric diseases, such as Parkinson`s disease, drug addiction, schizophrenia, depression and Huntington`s disease etc. In order to get more efficient, more sensitive CFE to detect DA release in brain, we improved traditional carbon glue-made CFE (C_CFE), and used ethylene glycol monothyl ether (EGME) as conduct material. Compared to C_CFE, EGME-made CFE (Y_CFE) is charaterised by fewer materials to be manufactured conveniently , more stable property of biology, and higher sensitivity, and can be applied to biological experiments. Our previous experiments showed that the complex firing pattern of action potential (AP) defined by a 4-parameter“code function”-F(n,m,f,d) effectively modulated neurotransmitter secretion in rat adrenal chromaffine cells (RACCs), where the four coding parameters (n,m,f,d) were defined as follows, a series of consecutive APs is defined as a “burst”; “n” is the number of APs in a burst; “m” is the number of bursts in the whole AP pattern; “f” is the frequency of APs in a burst; and “d” is the interval between two adjacent bursts (when m=1, “d” has no meaning).We found, in addition to AP frequency (f), for a given number of APs, another parameter of the AP code, the number of AP bursts (m) in which the set of APs occurs, effectively modulated cell secretion. However, it is not clear whether “m” effect can effectively modulate neurotransmitter release in vivo. In our study, we stimulated medial forebrain bundle of anaesthetized rats with different AP codes and recorded the real-time DA release in striatum with Y_CFE. On the other hand, we observed DA release in striatum slice, and nucleus accumbens, hippocampus and medial prefrontal cortex in vivo. Our work indicated that, Compared to C_CFE, EGME-made CFE (Y_CFE) is characterized by fewer materials to be manufactured conveniently , more stable property of biology, and higher sensitivity, and can be applied to biological experiments. Measurement of DA release showed that “m” effect effectively modulated DA release and were different with different AP codes in vivo.
Language: 中文
Content Type: 学位论文
Appears in Collections:神经所(总)_学位论文

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Recommended Citation:
大鼠纹状体多巴胺分泌的电化学记录技术及其在体(in vivo)研究.姚伟[d].中国科学院上海生命科学研究院,2007.20-25
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