The effects of alcohol on cerebellar synaotic transmission and plasticity

酒精对小脑突触传递和可塑性的影响

• 批准号: 7753907
• 负责人: Christian Robert Hansel
• 金额: $ 23.17万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-01-01 至 2011-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7753907
• 关键词: AMPA Receptors; Acute; Affect; Alcohol consumption; Alcohols; Area; Bathing; Brain; Calcium; Calcium Channel; Calcium Signaling; Calcium Spikes; Cause of Death; Cells; Cerebellar cortex structure; Cerebellum; Collaborations; Complex; Data; Death Rate; Dendrites; Dose; Ethanol; Excitatory Synapse; Fiber; Financial Support; Hand; Hippocampus (Brain); Image; Imaging Techniques; Impairment; In Vitro; Laboratories; Lead; Learning; Left; Location; Long-Term Depression; Long-Term Potentiation; Maps; Measures; Mediating; Memory; Metabotropic Glutamate Receptors; Mibefradil; Modeling; Monitor; Motor; Movement; N-Methyl-D-Aspartate Receptors; NMDA receptor antagonist; National Institute on Alcohol Abuse and Alcoholism; Neurons; New Mexico; Nimodipine; Organism; Output; Patch-Clamp Techniques; Phospholipase; Physiologic pulse; Physiology; Plastics; Play; Preparation; Probability; Process; Protein Kinase C; Protein phosphatase; Protocols documentation; Purkinje Cells; Rattus; Receptor Activation; Reporting; Research; Rest; Role; Signal Pathway; Signal Transduction; Slice; Specificity; Speed; Structure; Synapses; Synaptic Transmission; Synaptic plasticity; System; Techniques; Tilia; Time; Traffic accidents; Universities; Update; Work; alcohol effect; alcohol research; base; charge coupled device camera; data acquisition; design; environmental change; experience; innovation; interest; motor impairment; motor learning; novel; patch clamp; postsynaptic; presynaptic; public health relevance; receptor; receptor function; research study; response; tool; transmission process; voltage

DESCRIPTION (provided by applicant): Plasticity in brain motor systems allows fine adjustments of motor coordination that can develop over long periods of time, but can also occur within minutes. Rapid motor learning rests on synaptic plasticity at excitatory synaptic inputs to cerebellar Purkinje cells. Synaptic gain changes such as long-term potentiation (LTP) and long-term depression (LTD) at parallel fiber (PF) and climbing fiber (CF) synapses onto Purkinje cells provide a cellular basis for cerebellar motor learning. Alcohol is known to interfere with synaptic transmission and plasticity, but acute alcohol effects on cerebellar synaptic plasticity have not been studied so far, although it is well known that acute consequences of alcohol consumption include the impairment of motor coordination and the ability to fine-tune movements. Here, we propose to examine the effects of acute ethanol application on cerebellar synaptic plasticity in rat brain slices using whole-cell patch-clamp recordings as well as microfluorometric imaging techniques. Aside from our preliminary experiments performed in preparation for this application, we have no previous experience in alcohol-related research. However, we have characterized cellular mechanisms underlying cerebellar synaptic plasticity (e.g. Hansel et al., 2001; Jvrntell and Hansel, 2006). This application for financial support from the National Institute on Alcohol Abuse and Alcoholism (NIAAA) is motivated by our hope that we now have the required experience and tools at hand to study how ethanol interferes with cerebellar learning mechanisms. Our first steps into the field of alcohol research will be facilitated by a collaboration with the laboratory of Prof. C.F. Valenzuela (University of New Mexico at Albuquerque, USA), who has long-standing experience in this field. Here, we suggest three specific aims: a) to examine acute ethanol effects on cerebellar synaptic transmission and synaptic plasticity, b) to examine ethanol effects on NMDA receptors, metabotropic glutamate receptors and voltage-dependent calcium channels in Purkinje cells and c) to use calcium imaging techniques, in combination with somato-dendritic patch-clamp techniques, to monitor ethanol effects on the spatio-temporal map of dendritic calcium spikes in Purkinje cell dendrites. These dendritic calcium spikes, which are crucial for cerebellar plasticity, are likely triggered by voltage-gated calcium currents and NMDA receptors. Both have been described as ethanol targets in other types of neurons, and therefore dendritic calcium spikes might be affected by ethanol as well. Our proposal has an innovative and exploratory character, as we plan to use a novel combination of imaging (ultra high-speed calcium imaging) and patch-clamp techniques (triple patching with one somatic and two dendritic locations) to be able to characterize dendritic calcium spike activity, which has not been done so far in alcohol research. We believe that this type of research is important to better understand the cellular basis of acute effects of alcohol consumption. One of these is impaired motor coordination, which contributes to the high rate of deaths caused by alcohol-related traffic accidents. PUBLIC HEALTH RELEVANCE: The cerebellum is a brain structure involved in the fine-adjustment of motor coordination and in motor learning. These processes can be impaired as a consequence of acute alcohol consumption, and yet acute alcohol effects on cerebellar synaptic transmission and on plasticity within cerebellar networks have not been studied so far. Here, we propose to examine alcohol effects on forms of cerebellar synaptic plasticity as well as the cellular causes for such effects using electrophysiological recording techniques and fluorometric calcium imaging techniques in rat brain slices.

描述(申请人提供)：大脑运动系统的可塑性允许运动协调的微调，这可能会在很长一段时间内发展，但也可能在几分钟内发生。快速运动学习依赖于小脑浦肯野细胞兴奋性突触输入的突触可塑性。平行纤维(PF)的长时程增强(LTP)和长时程抑制(LTD)以及浦肯野细胞的爬升纤维(CF)突触等突触增益变化为小脑运动学习提供了细胞学基础。酒精干扰突触传递和可塑性是已知的，但急性酒精对小脑突触可塑性的影响到目前为止还没有研究，尽管众所周知，饮酒的严重后果包括运动协调性和微调运动的能力受损。在这里，我们建议使用全细胞膜片钳记录和显微荧光成像技术来研究急性乙醇应用对大鼠脑片小脑突触可塑性的影响。除了我们为准备这项应用而进行的初步实验外，我们以前没有酒精相关研究的经验。然而，我们已经描述了小脑突触可塑性的细胞机制(例如Hansel等人，2001；Jvrntell和Hansel，2006)。这次申请国家酒精滥用和酒精中毒研究所(NIAAA)的财政支持是因为我们希望现在我们手头有必要的经验和工具来研究乙醇如何干扰小脑学习机制。我们进入酒精研究领域的第一步将通过与C.F.Valenzuela教授(美国新墨西哥大学阿尔伯克基分校)的实验室合作来推动，他在该领域拥有长期经验。在这里，我们提出了三个特定的目标：a)检测急性酒精对小脑突触传递和突触可塑性的影响；b)检测酒精对浦肯野细胞NMDA受体、代谢性谷氨酸受体和电压依赖性钙通道的影响；c)结合体细胞-树突状膜片钳技术，监测乙醇对浦肯野细胞树突中树突状钙峰时空图的影响。这些树突状钙峰对小脑的可塑性至关重要，可能是由电压门控钙电流和NMDA受体触发的。在其他类型的神经元中，两者都被描述为乙醇靶点，因此树突钙峰可能也会受到乙醇的影响。我们的建议具有创新性和探索性，因为我们计划使用成像(超高速钙成像)和膜片钳技术(一个体细胞和两个树突位置的三重贴片)的新组合来表征树突状钙峰活动，这在酒精研究中迄今尚未完成。我们认为，这种类型的研究对于更好地理解饮酒急性影响的细胞基础是重要的。其中之一是运动协调性受损，这是与酒精有关的交通事故导致的高死亡率的原因之一。 公共卫生相关性：小脑是参与运动协调和运动学习的微调的大脑结构。这些过程可能会因为急性饮酒而受损，但到目前为止，急性酒精对小脑突触传递和小脑网络可塑性的影响还没有研究过。在这里，我们建议使用大鼠脑片的电生理记录技术和荧光钙成像技术来研究酒精对小脑突触可塑性形式的影响以及这种影响的细胞原因。

项目成果

Purkinje cell NMDA receptors assume a key role in synaptic gain control in the mature cerebellum.

• DOI: 10.1523/jneurosci.4344-10.2010
• 发表时间: 2010-11-10
• 期刊: The Journal of neuroscience : the official journal of the Society for Neuroscience
• 作者: Piochon C;Levenes C;Ohtsuki G;Hansel C
• 通讯作者: Hansel C