阿拉巴马大学伯明翰学院发展资助计划提供支持。

<html> 1。心理生理学 BIOPAC系统，旗下的生理监测系统（参见具体设备表），非标设备在大多数成像设施。附表15-30分钟参与者的到来之前设立的生理监测和本议定书中所描述的其他设备（图1）。 <ol><li>连接控制室的电脑操作系统AcqKnowledge（BIOPAC系统公司）生理监测软件使用标准的以太网交叉电缆（CBLETH2）BIOPAC MP150（MP150WSW）。 </li><li> BIOPAC隔离数字接口（STP100C）连接到控制室的电脑操作演示（神经行为系统，INC;奥尔巴尼，加利福尼亚）软件，采用一个DB25的男/女带状电缆。 </li><li>连接BIOPAC金沙江放大器（EDA - 100C - MRI）的射频干扰滤波器（MRIRFIF），使用屏蔽延长线（MECMRI 3）内部控制室。 </li><li>连接射频干扰的屏蔽延长线滤波器（MRIRFIF）（MECMRI 1）在MRI扫描室。 </li><li>碳纤维导线（铅108）连接到无线电半透明电极（EL508）屏蔽延长线连接。注:在紧张的螺旋捻的线索减少工件在皮肤电导，可以在扫描过程中创建的数据。 </li><li>将无线电半透明电极（EL508）参与者的左手中指和无名指末节。 </li><li>由于扫描设备的性质，核磁共振室的室内温度往往低于21 ° C封面用毛毯，以保持手的温度的参与者。 </li></ol> 2。行为反应（操纵杆） <ol><li>操纵杆的fORP接口单元（目前的设计公司，费城，PA）使用迷你USB电缆连接控制室的计算机操作演示软件（神经行为系统，INC;奥尔巴尼，加利福尼亚）。 </li><li>连接光缆在控制室fORP接口单元，然后通过波导到核磁共振室的电缆。 </li><li>光缆连接致辞兼容的操纵杆。 </li><li>的直接参与者，放置在一个舒适和容易到达的位置操纵杆。 </li></ol> 3。刺激呈现<ol><li>国际金融机构SA（Invivo公司，佛罗里达州奥兰多市）控制室控制台（图1）外部VGA和音频端口连接控制室的计算机操作演示软件。 </li><li>检查国际金融机构控制室控制台和国际金融机构的外设接口单元内MRI室，以及外围接口单元和音频/视频显示单元之间的连接之间的光纤电缆连接。 </li><li>将头线圈等，参与者可以通过连接到头部线圈的镜子查看显示器背后的音频/视频显示单元。 </li><li>国际金融机构系统的MR兼容立体声耳机，使用乙烯管，连接的音频/视频显示单元的声学接口盒。 </li><li>校准使用声压级米的听觉刺激量。 </li></ol> 4。实验过程<ol><li>通知与会者2铃声将在研究过程中几次，铃声的音量会有所不同的上方和下方的感知阈值（图2）。 </li><li>直接参与者，立即推动操纵杆盒上的一个按钮后，听证会或者音，然后更新他们的期望，通过移动操纵杆来控制一个在0至100分的评分栏的位置（图3）接收的UCS。 </li><li>指示参加他们的UCS的预期率在连续的规模从0到100。告知他们的0个评分表明他们一定不会提出的UCS，50的收视率表明，他们是不确定的UCS是否将提交，和100的收视率表明，他们有一定的UCS将提交。直接参与者使用上规模的其他值来表示中间的期望。然后，让学员练习使用操纵杆，使收视率。 </li><li>让参与者差分恐惧制约的程序，使用2个音（700和1300赫兹，20多岁的ITI的10秒持续时间）作为条件刺激（CS）和一个响亮的白噪声（100分贝500毫秒）作为UCS。 </li><li>目前60试验的CS +（coterminating与UCS）和60项试验的CS -（没有的UCS）一个伪秩序等，呈现连续不超过2试验相同的CS。 </li><li>抵消作为CS +和CS -跨参与者的色调。 </li><li>调节的CS +和CS独立的音量。调整后续试验相同的CS CS量。如果按下一个按钮（即认为试验），减少CS量5分贝。如果没有按下一个按钮增加音量5分贝（即以下unperceived审判）。 </li></ol><html> ve_title"扫描程序&gt; 5。 <ol><li>收集T1加权标准的高分辨率结构图像（例如MPRAGE）作为功能数据的解剖参考。 </li><li>收集大胆在调节过程中的全脑功能磁共振成像。三十六，4mm厚的片，应足以弥补相对标准的成像参数（如TR = 2000毫秒，TE = 30ms的，FOV = 24厘米，64x64的矩阵）的大脑。刺激呈现同步使用的功能磁共振成像触发盒的功能磁共振成像收购。 </li></ol> 6。 SCR的数据采集和分析<ol><li>样品在2000赫兹，使用AcqKnowledge软件和MR兼容BIOPAC生理监测系统，在第1条所述的皮肤电导。 </li><li>应用1 Hz的无限脉冲响应（IIR）的皮肤电导数据低通数字滤波器，以减少在成像产生的工件（见图4）。 </li><li>重新取样，在250赫兹的皮肤电导数据。 </li><li>皮肤电导响应发病水平的差异，以响应峰值计算的SCR。 </li><li> SCR的数据可以转化为规范化的响应振幅分布统计分析前的平方根。 </li></ol> 7。 UCS的预期数据采集与分析<ol><li>样品（40赫兹）和记录UCS预期的数据，使用演示软件。 </li><li>计算平均响应（1S样品）在最后一秒的CS演示UCS的预期。 </li></ol> 8。功能磁共振成像数据采集与分析<ol><li>完整的标准预处理的脑成像数据，使用功能成像分析软件（如AFNI 18）（如切片定时校正，图像配准，空间平滑） 。 </li><li>创建标准滋扰（如运动）和刺激的CS +和CS -，以及UCS的感知和unperceived试验基于回归。 </li><li>创建一个电机响应的参考波形作为一个滋扰回归量电机活动相关按下按钮响应。 <ol class="lalpha"><li>创建一棒功能，​​按下按钮响应时间的代码。 </li><li>卷积与规范的血流动力学响应函数（HRF）按下按钮坚持功能。 </li></ol></li><li>创建电机响应的参考波形作为一个滋扰回归量操纵杆反应有关的运动活动。 <ol class="lalpha"><li>创建一棒功能，​​在斜坡UCS的预期收视率的变化（如坡度绝对值&gt; 10）的时间代码。 </li><li>卷积操纵杆斜坡坚持规范HRF功能。 </li></ol></li><li>执行第一层次的分析，利用一切刺激和滋扰回归。 </li><li>执行第二个层次的重复测量方差分析，以确定激活其中的一个观念的主要作用，CS型或CS X型感知互动的主要影响的地区。 </li></ol> 9。代表性的成果: 这里介绍的方法UCS的预期收视率相对较高，在通常认为CS +试验，并在认为CS -试验（图5）10,15,19收视率低的结果。这些结果表明，参与者都知道CS - UCS的突发。在unperceived试验，UCS的预期收视率通常保持预CS评级不变。通常在这些unperceived CS +和CS -试验UCS预期下降近50个说明参与者是不确定的UCS是否将提交 10,15,19（图5） 。这无法产生UCS的预期收视率差的unperceived CS +和CS -表示，参与者都无法表达的应变意识unperceived空调试验（图6）。相比之下，学习相关的变化，在SCR已经观察到在感知和unperceived空调试验 10,15,19 。具体来说，可控硅均大于的感知，感知的CS + CS -。同样的，更大的可控硅unperceived政务司司长期间已被证明+比unperceived CS -试验10,15,19（图6）。两者合计，这些行为和自主的数据表明，没有应急意识unperceived试验认为试验与应急意识，恐惧条件和恐惧条件。功能成像研究，使用这种方法已经证明对知觉学习相关的海马激活，但不unperceived空调试验 15（图7） 。相反，差的杏仁核活动是观察感知和unperceived空调试验 15 。这些研究结果是一致认为海马的支持与应急意识的过程，而杏仁核华润表达支持与不认识。 / 3083/3083fig1.jpg"ALT ="图1"/&gt; 图1图的基本设备，为刺激呈现和行为/心理生理反应监测。演示软件是用于目前的视听刺激和监视UCS的预期收视率，用右手移动操纵杆。 AcqKnowledge软件和BIOPAC设备用来监视从左侧皮肤电导。固体（BIOPAC），单虚线（IFIS音像），双虚线线（光纤操纵杆）描绘了不同的刺激呈现和反应监测系统的电缆。黑色箭头表示信息流的方向。 <img src="/files/ftp_upload/3083/3083fig2.jpg" alt="图2"/> 图2:空调的刺激。目前CS +和CS -一个伪秩序等，呈现连续不超过2试验相同的CS。改变CS +和CS -独立音量。如果CS被视为（按下一个按钮表示），减少对随后的审判相同的CS CS量5分贝。如果CS是unperceived（表示没有按下按钮），提高CS量5分贝相同的CS随后的审讯。 <img src="/files/ftp_upload/3083/3083fig3.jpg" alt="图3"/> 图3。UCS预期评级规模。指示参加率在0至100分的期望UCS的介绍。评分为0表示肯定不会提出的UCS，100的收视率表明确定性的UCS将提交，和50的评级反映了是否将提交的UCS的不确定性。中级评分应该用来指示UCS的预期层次。 <img src="/files/ftp_upload/3083/3083fig4.jpg" alt="图4"/> 图4。原材料和过滤的皮肤电导数据的比较。一）原料皮电导数据收集过程中的fMRI。 b）皮肤电导数据后1Hz的IIR低通滤波器的应用程序。 <img src="/files/ftp_upload/3083/3083fig5.jpg" alt="图5"/> 图5。UCS预期收视率。通常，与会者报告认为CS +试验和低知觉CS试验预期高UCS的预期。在unperceived CS +和CS试验UCS预期没有什么不同。 <img src="/files/ftp_upload/3083/3083fig6.jpg" alt="图6"/> 图6。UCS的预期和SCR。 UCS的预期的差异通常认为CS +和CS试验观察表明参与者都知道刺激应急。 unperceived试验，UCS的预期收视率通常不有所不同，参与者都无法表达自己的应急意识。相比之下，调节可控硅的差异通常是观察认识和unperceived空调试验。这样的结果反映（即认为试验）和无（unperceived试验IE）应急意识了解到恐惧的表达。 <img src="/files/ftp_upload/3083/3083fig7.jpg" alt="图7"/> 图7。海马和杏仁核的功能磁共振成像。海马的反应通常大于CS -感知，但没有unperceived空调试验的CS +。差分杏仁核的反应通常是感知和unperceived空调试验观察。这些研究结果是一致认为海马的支持与应急意识的过程，而杏仁核恐惧表达支持与不认识。

<ol>
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没有利益冲突的声明。

这里描述的恐惧调理方法调查知道和不知道的恐惧记忆过程的神经机制提供了一种手段。这种方法需要的行为，植物神经和fMRI数据的同步监测的优势。监测行为（即UCS的预期）和植物神经反应（即可控硅）是这种方法的关键组成部分。 UCS的预期提供了一种手段，以评估应变意识，而SCR CR的表达提供了一个指数。总之，这些行为和植物神经反应，可用于在超阈CS +和CS试验演示调查恐惧条件和没有?...

<table border="1"><tbody><tr><td>设备</td><td>公司</td><td>编号</td></tr><tr><td>综合功能成像系统（IFIS - SA） </td><td> Invivo公司，佛罗里达州奥兰多市</td><td></td></tr><tr><td>主控制单元（位于中控室） 外围接口单元（设在MRI室） 音频/视频显示单元（位于在MRI室），包括: <ul><li> 6.4"（对角线）的LCD屏幕<ul><li> 640 x 480分辨率和15 °的角度场</li></ul></li><li>声学接口盒<ul><li>提供气动立体声音响</li></ul></li><li> MR兼容立体声耳机</li></ul></td><td></td><td></td></tr><tr><td>生理监测系统</td><td> BIOPAC系统公司，Goleta，CA </td><td></td></tr><tr><td>数据采集和分析系统为Windows（MP150） 隔离数字接口（ 数字接口 ） 皮电反应（GSR）放大器 MRI有线/传感器放大器设置的过滤系统，包括: <ul><li> MRI延长线（商会过滤器） </li><li>射频干扰滤波器</li><li> MRI延长线（金沙江放大器来过滤） </li></ul> 附加组件: DB25的男/女带状电缆 一次性radiotranslucent电极 碳纤维引线</td><td></td><td valign="top"> MP150WSW STP100C EDA100C - MRI MECMRI，反 - MECMRI - 1 - MRIRFIF - MECMRI - 3 CBL110C EL508 LEAD108 </td></tr><tr><td>操纵杆</td><td>目前的设计公司，宾夕法尼亚州费城</td><td></td></tr><tr><td>传统摇杆</td><td></td><td> HH - JOY - 4 </td></tr><tr><td>传统fORP接口</td><td></td><td>金融情报机构- 005 </td></tr></tbody></table>

investigating the neural mechanisms of aware and unaware fear memory with fmri

巴甫洛夫担心空调往往是结合使用功能磁共振成像（fMRI）技术，在人类联想学习1-5的神经基础研究。在这些研究中，重要的是要提供空调的行为的证据，以验证是学习相关的，与人类行为相关的大脑活动的差异。 恐惧条件的研究往往监察植物神经反应（如皮肤电导反应; SCR）作为学习和记忆 6-8的索引。此外，其他行为的措施，可以提供有价值的信息关注学习过程和/或其他认知功能的影响空调。例如，影响无条件刺激（UCS）的预期有上表达的条件反应（CR）和无条件反应（UCR）数9-14最近的研究已经在一个感兴趣的话题。 SCR和UCS预期措施最近已与功能磁共振成像结合调查知道和不知道害怕学习和记忆过程15的神经基础。虽然这些认知过程可以评估在一定程度以下的空调会议，评估后空调不能衡量审判试行的期望和易受干扰和遗忘，以及其他因素，可能会扭曲结果16,17 。 自主和行为反应，同时监测功能磁共振成像提供了一个调解的认知过程和行为/植物神经反应之间的复杂关系的神经基板可以评估机制。然而，在MRI环境监测的自主和行为反应，对一些实际问题。具体来说，1）标准的行为和生理监测设备是构建有色金属材料，可以不被安全地附近的磁共振成像扫描仪，2使用）时，此设备是以外的磁共振扫描室放置，投射到主体的电缆可以进行射频噪声，文物在脑图像，3）文物可产生皮肤内的电导信号开关在扫描过程中，4梯度）的行为反应的电机要求所产生的fMRI信号，可能需要从活动区别开来，涉及到利益的认知过程。这些问题都可以解决与生理监测设备的安装和额外的数据分析程序进行修改。在这里，我们提出了一个方法同时监测在功能磁共振成像的自主和行为反应，并证明这些方法的使用调查期间恐惧制约知道和不知道的记忆过程。

Watch this Scientific Journal Video about Investigating the Neural Mechanisms of Aware and Unaware Fear Memory with fMRI at JoVE.com

Investigating the Neural Mechanisms of Aware and Unaware Fear Memory with fMRI

描述的方法进行调查知道和不知道的记忆过程的神经机制，支持在恐惧条件。这种方法监测血氧水平依赖（BOLD）的功能磁共振成像，皮肤电导反应，和无条件刺激的预期，在巴甫洛夫恐惧的调节，以评估鲜明的记忆过程的神经关联。

调查的意识到，不知道用功能磁共振成像的恐惧记忆的神经机制

Pavlovian fear conditioning is often used in combination with functional magnetic resonance imaging (fMRI) in humans to investigate the neural substrates ...

investigating-neural-mechanisms-aware-unaware-fear-memory-with

Research

JoVE Journal

Neuroscience

12.8K 次观看.  University of Alabama at Birmingham. 描述的方法进行调查知道和不知道的记忆过程的神经机制，支持在恐惧条件。这种方法监测血氧水平依赖（BOLD）的功能磁共振成像，皮肤电导反应，和无条件刺激的预期，在巴甫洛夫恐惧的调节，以评估鲜明的记忆过程的神经关联。

视频: Investigating the Neural Mechanisms of Aware and Unaware Fear Memory with fMRI

Simultaneous fMRI and Electrophysiology in the Rodent Brain

To examine the neural basis of the blood oxygenation level dependent (BOLD) magnetic resonance imaging (MRI) signal, we have developed a rodent model in which functional MRI data and in vivo intracortical recording can be performed simultaneously. The combination of MRI and electrical recording is technically challenging because the electrodes used for recording distort the MRI images and the MRI acquisition induces noise in the electrical recording. To minimize the mutual interference of the two modalities, glass microelectrodes were used rather than metal and a noise removal algorithm was implemented for the electrophysiology data. In our studies, two microelectrodes were separately implanted in bilateral primary somatosensory cortices (SI) of the rat and fixed in place. One coronal slice covering the electrode tips was selected for functional MRI. Electrode shafts and fixation positions were not included in the image slice to avoid imaging artifacts. The removed scalp was replaced with toothpaste to reduce susceptibility mismatch and prevent Gibbs ringing artifacts in the images. The artifact structure induced in the electrical recordings by the rapidly-switching magnetic fields during image acquisition was characterized by averaging all cycles of scans for each run. The noise structure during imaging was then subtracted from original recordings. The denoised time courses were then used for further analysis in combination with the fMRI data. As an example, the simultaneous acquisition was used to determine the relationship between spontaneous fMRI BOLD signals and band-limited intracortical electrical activity. Simultaneous fMRI and electrophysiological recording in the rodent will provide a platform for many exciting applications in neuroscience in addition to elucidating the relationship between the fMRI BOLD signal and neuronal activity.

We have developed a method for simultaneous functional magnetic resonance imaging and electrophysiological recording in the rodent brain, providing a platform for the investigation of the relationship between neural activity and the blood oxygenation level dependent (BOLD) MRI signal.

同时在啮齿动物脑功能磁共振成像和电

To examine the neural basis of the blood oxygenation level dependent (BOLD) magnetic resonance imaging (MRI) signal, we have developed a rodent model in ...

科研

神经科学

Physiological, Morphological and Neurochemical Characterization of Neurons Modulated by Movement

The role of individual neurons and their function in neuronal circuits is fundamental to understanding the neuronal mechanisms of sensory and motor functions. Most investigations of sensorimotor mechanisms rely on either examination of neurons while an animal is static1,2 or record extracellular neuronal activity during a movement.3,4 While these studies have provided the fundamental background for sensorimotor function, they either do not evaluate functional information which occurs during a movement or are limited in their ability to fully characterize the anatomy, physiology and neurochemical phenotype of the neuron. A technique is shown here which allows extensive characterization of individual neurons during an in vivo movement. This technique can be used not only to study primary afferent neurons but also to characterize motoneurons and sensorimotor interneurons. Initially the response of a single neuron is recorded using electrophysiological methods during various movements of the mandible followed by determination of the receptive field for the neuron. A neuronal tracer is then intracellularly injected into the neuron and the brain is processed so that the neuron can be visualized with light, electron or confocal microscopy (Fig. 1). The detailed morphology of the characterized neuron is then reconstructed so that neuronal morphology can be correlated with the physiological response of the neuron (Figs. 2,3). In this communication important key details and tips for successful implementation of this technique are provided. Valuable additional information can be determined for the neuron under study by combining this method with other techniques. Retrograde neuronal labeling can be used to determine neurons with which the labeled neuron synapses; thus allowing detailed determination of neuronal circuitry. Immunocytochemistry can be combined with this method to examine neurotransmitters within the labeled neuron and to determine the chemical phenotypes of neurons with which the labeled neuron synapses. The labeled neuron can also be processed for electron microscopy to determine the ultrastructural features and microcircuitry of the labeled neuron. Overall this technique is a powerful method to thoroughly characterize neurons during in vivo movement thus allowing substantial insight into the role of the neuron in sensorimotor function.

A technique is described to quantify the in vivo physiological response of mammalian neurons during movement and correlate the physiology of the neuron with neuronal morphology, neurochemical phenotype and synaptic microcircuitry.

由运动神经元调制生理，形态学和神经化学的表征

The role of individual neurons and their function in neuronal circuits is fundamental to understanding the neuronal mechanisms of sensory and motor ...

Extinction Training During the Reconsolidation Window Prevents Recovery of Fear

Fear is maladaptive when it persists long after circumstances have become safe. It is therefore crucial to develop an approach that persistently prevents the return of fear. Pavlovian fear-conditioning paradigms are commonly employed to create a controlled, novel fear association in the laboratory. After pairing an innocuous stimulus (conditioned stimulus, CS) with an aversive outcome (unconditioned stimulus, US) we can elicit a fear response (conditioned response, or CR) by presenting just the stimulus alone1,2 . Once fear is acquired, it can be diminished using extinction training, whereby the conditioned stimulus is repeatedly presented without the aversive outcome until fear is no longer expressed3. This inhibitory learning creates a new, safe representation for the CS, which competes for expression with the original fear memory4. Although extinction is effective at inhibiting fear, it is not permanent. Fear can spontaneously recover with the passage of time. Exposure to stress or returning to the context of initial learning can also cause fear to resurface3,4.
Our protocol addresses the transient nature of extinction by targeting the reconsolidation window to modify emotional memory in a more permanent manner. Ample evidence suggests that reactivating a consolidated memory returns it to a labile state, during which the memory is again susceptible to interference5-9. This window of opportunity appears to open shortly after reactivation and close approximately 6hrs later5,11,16, although this may vary depending on the strength and age of the memory15. By allowing new information to incorporate into the original memory trace, this memory may be updated as it reconsolidates10,11. Studies involving non-human animals have successfully blocked the expression of fear memory by introducing pharmacological manipulations within the reconsolidation window, however, most agents used are either toxic to humans or show equivocal effects when used in human studies12-14. Our protocol addresses these challenges by offering an effective, yet non-invasive, behavioral manipulation that is safe for humans.
By prompting fear memory retrieval prior to extinction, we essentially trigger the reconsolidation process, allowing new safety information (i.e., extinction) to be incorporated while the fear memory is still susceptible to interference. A recent study employing this behavioral manipulation in rats has successfully blocked fear memory using these temporal parameters11. Additional studies in humans have demonstrated that introducing new information after the retrieval of previously consolidated motor16, episodic17, or declarative18 memories leads to interference with the original memory trace14. We outline below a novel protocol used to block fear recovery in humans.

Conditioned fear can be diminished through an inhibitory process called extinction, but can resurface under conditions such as the passage of time or exposure to stress. Our protocol presents a novel way of preventing fear recovery by introducing extinction during the reconsolidation window (the re-storage phase of a reactivated memory).

灭绝培训在彭兰霞窗口无法还原的恐惧

Fear is maladaptive when it persists long after circumstances have become safe. It is therefore crucial to develop an approach that persistently prevents ...

Optical Recording of Suprathreshold Neural Activity with Single-cell and Single-spike Resolution

Signaling of information in the vertebrate central nervous system is often carried by populations of neurons rather than individual neurons. Also propagation of suprathreshold spiking activity involves populations of neurons. Empirical studies addressing cortical function directly thus require recordings from populations of neurons with high resolution. Here we describe an optical method and a deconvolution algorithm to record neural activity from up to 100 neurons with single-cell and single-spike resolution. This method relies on detection of the transient increases in intracellular somatic calcium concentration associated with suprathreshold electrical spikes (action potentials) in cortical neurons. High temporal resolution of the optical recordings is achieved by a fast random-access scanning technique using acousto-optical deflectors (AODs)1. Two-photon excitation of the calcium-sensitive dye results in high spatial resolution in opaque brain tissue2. Reconstruction of spikes from the fluorescence calcium recordings is achieved by a maximum-likelihood method. Simultaneous electrophysiological and optical recordings indicate that our method reliably detects spikes (&gt;97% spike detection efficiency), has a low rate of false positive spike detection (&lt; 0.003 spikes/sec), and a high temporal precision (about 3 msec) 3. This optical method of spike detection can be used to record neural activity in vitro and in anesthetized animals in vivo3,4.

Understanding the function of the vertebrate central nervous system requires recordings from many neurons because cortical function arises on the level of populations of neurons. Here we describe an optical method to record suprathreshold neural activity with single-cell and single-spike resolution, dithered random-access scanning. This method records somatic fluorescence calcium signals from up to 100 neurons with high temporal resolution. A maximum-likelihood algorithm deconvolves the underlying suprathreshold neural activity from the somatic fluorescence calcium signals. This method reliably detects spikes with high detection efficiency and a low rate of false positives and can be used to study neural populations in vitro and in vivo.

阈上的神经活动的单细胞和单穗的分辨率的光学的方法记录

Signaling of information in the vertebrate central nervous system is often carried by populations of neurons rather than individual neurons. Also ...

Co-analysis of Brain Structure and Function using fMRI and Diffusion-weighted Imaging

The study of complex computational systems is facilitated by network maps, such as circuit diagrams. Such mapping is particularly informative when studying the brain, as the functional role that a brain area fulfills may be largely defined by its connections to other brain areas. In this report, we describe a novel, non-invasive approach for relating brain structure and function using magnetic resonance imaging (MRI). This approach, a combination of structural imaging of long-range fiber connections and functional imaging data, is illustrated in two distinct cognitive domains, visual attention and face perception. Structural imaging is performed with diffusion-weighted imaging (DWI) and fiber tractography, which track the diffusion of water molecules along white-matter fiber tracts in the brain (Figure 1). By visualizing these fiber tracts, we are able to investigate the long-range connective architecture of the brain. The results compare favorably with one of the most widely-used techniques in DWI, diffusion tensor imaging (DTI). DTI is unable to resolve complex configurations of fiber tracts, limiting its utility for constructing detailed, anatomically-informed models of brain function. In contrast, our analyses reproduce known neuroanatomy with precision and accuracy. This advantage is partly due to data acquisition procedures: while many DTI protocols measure diffusion in a small number of directions (e.g., 6 or 12), we employ a diffusion spectrum imaging (DSI)1, 2 protocol which assesses diffusion in 257 directions and at a range of magnetic gradient strengths. Moreover, DSI data allow us to use more sophisticated methods for reconstructing acquired data. In two experiments (visual attention and face perception), tractography reveals that co-active areas of the human brain are anatomically connected, supporting extant hypotheses that they form functional networks. DWI allows us to create a "circuit diagram" and reproduce it on an individual-subject basis, for the purpose of monitoring task-relevant brain activity in networks of interest.

We describe a novel approach for simultaneous analysis of brain function and structure using magnetic resonance imaging (MRI). We assess brain structure with high-resolution diffusion-weighted imaging and white-matter fiber tractography. Unlike standard structural MRI, these techniques allow us to directly relate anatomical connectivity to functional properties of brain networks.

使用功能磁共振成像和弥散加权成像脑结构与功能分析

The study of complex computational systems is facilitated by network maps, such as circuit diagrams. Such mapping is particularly informative when ...

Whole Cell Patch Clamp for Investigating the Mechanisms of Infrared Neural Stimulation

It has been demonstrated in recent years that pulsed, infrared laser light can be used to elicit electrical responses in neural tissue, independent of any further modification of the target tissue. Infrared neural stimulation has been reported in a variety of peripheral and sensory neural tissue in vivo, with particular interest shown in stimulation of neurons in the auditory nerve. However, while INS has been shown to work in these settings, the mechanism (or mechanisms) by which infrared light causes neural excitation is currently not well understood. The protocol presented here describes a whole cell patch clamp method designed to facilitate the investigation of infrared neural stimulation in cultured primary auditory neurons. By thoroughly characterizing the response of these cells to infrared laser illumination in vitro under controlled conditions, it may be possible to gain an improved understanding of the fundamental physical and biochemical processes underlying infrared neural stimulation.

Infrared nerve stimulation has been proposed as an alternative to electrical stimulation in a range of nerve types, including those associated with the auditory system. This protocol describes a patch clamp method for studying the mechanism of infrared nerve stimulation in a culture of primary auditory neurons.

全细胞膜片钳红外神经刺激机制的调查

It has been  demonstrated in recent years that pulsed, infrared laser light can be used to  elicit electrical responses in neural tissue, independent of ...

Optogenetic Perturbation of Neural Activity with Laser Illumination in Semi-intact Drosophila Larvae in Motion

Drosophila larval locomotion is a splendid model system in developmental and physiological neuroscience, by virtue of the genetic accessibility of the underlying neuronal components in the circuits1-6. Application of optogenetics7,8 in the larval neural circuit allows us to manipulate neuronal activity in spatially and temporally patterned ways9-13. Typically, specimens are broadly illuminated with a mercury lamp or LED, so specificity of the target neurons is controlled by binary gene expression systems such as the Gal4-UAS system14,15. In this work, to improve the spatial resolution to "sub-genetic resolution", we locally illuminated a subset of neurons in the ventral nerve cord using lasers implemented in a conventional confocal microscope. While monitoring the motion of the body wall of the semi-intact larvae, we interactively activated or inhibited neural activity with channelrhodopsin16,17 or halorhodopsin18-20, respectively. By spatially and temporally restricted illumination of the neural tissue, we can manipulate the activity of specific neurons in the circuit at a specific phase of behavior. This method is useful for studying the relationship between the activities of a local neural assembly in the ventral nerve cord and the spatiotemporal pattern of motor output.

Optogenetic Perturbation of Neural Activity with Laser Illumination in Semi-intact Drosophila Larvae in Motion

Here we describe a protocol for optogenetic manipulation of motoneuronal activity while monitoring changes in motor output (muscle contraction) in semi-intact Drosophila larvae using lasers within a conventional confocal microscope. This technique enables researchers to achieve local perturbation of neural activity in a few neuromeres to elucidate the dynamics of motor circuits.

光遗传学的扰动激光照射半完好的神经活动<em&gt;果蝇</em&gt;幼虫运动

Drosophila larval locomotion is a splendid model system in developmental and physiological neuroscience, by virtue of the genetic accessibility of the ...

Whole Mount Immunolabeling of Olfactory Receptor Neurons in the Drosophila Antenna

Odorant molecules bind to their target receptors in a precise and coordinated manner. Each receptor recognizes a specific signal and relays this information to the brain. As such, determining how olfactory information is transferred to the brain, modifying both perception and behavior, merits investigation. Interestingly, there is emerging evidence that cellular transduction and transcriptional factors are involved in the diversification of olfactory receptor neuron. Here we provide a robust whole mount immunological labeling method to assay in vivo olfactory receptor neuron organization. Using this method, we identified all olfactory receptor neurons with anti-ELAV antibody, a known pan-neural marker and Or49a-mCD8::GFP, an olfactory receptor neuron specifically expressed in Nba neuron using anti-GFP antibody.

Whole Mount Immunolabeling of Olfactory Receptor Neurons in the Drosophila Antenna

Herein we describe the process of whole mount immunostaining of Drosophila antennae, which enables us to better understand the molecular mechanisms involved in the diversification of olfactory receptor neurons (ORN)s.

嗅觉受体神经元在全山免疫标记<em&gt;果蝇</em&gt;天线

Odorant molecules bind to their target receptors in a precise and coordinated manner. Each receptor recognizes a specific signal and relays this ...

Deep Brain Stimulation with Simultaneous fMRI in Rodents

In order to visualize the global and downstream neuronal responses to deep brain stimulation (DBS) at various targets, we have developed a protocol for using blood oxygen level dependent (BOLD) functional magnetic resonance imaging (fMRI) to image rodents with simultaneous DBS. DBS fMRI presents a number of technical challenges, including accuracy of electrode implantation, MR artifacts created by the electrode, choice of anesthesia and paralytic to minimize any neuronal effects while simultaneously eliminating animal motion, and maintenance of physiological parameters, deviation from which can confound the BOLD signal.&#160;Our laboratory has developed a set of procedures that are capable of overcoming most of these possible issues. For electrical stimulation, a homemade tungsten bipolar microelectrode is used, inserted stereotactically at the stimulation site in the anesthetized subject. In preparation for imaging, rodents are fixed on a plastic headpiece and transferred to the magnet bore. For sedation and paralysis during scanning, a cocktail of dexmedetomidine and pancuronium is continuously infused, along with a minimal dose of isoflurane; this preparation minimizes the BOLD ceiling effect of volatile anesthetics. In this example experiment, stimulation of the subthalamic nucleus (STN) produces BOLD responses which are observed primarily in ipsilateral cortical regions, centered in motor cortex. Simultaneous DBS and fMRI allows the unambiguous modulation of neural circuits dependent on stimulation location and stimulation parameters, and permits observation of neuronal modulations free of regional bias. This technique may be used to explore the downstream effects of modulating neural circuitry at nearly any brain region, with implications for both experimental and clinical DBS.

This protocol describes a standard method for simultaneous functional magnetic resonance imaging and deep brain stimulation in the rodent. The combined use of these experimental tools allows for the exploration of global downstream activity in response to electrical stimulation at virtually any brain target.

深部脑刺激与同步功能磁共振成像在鼠害

In order to visualize the global and downstream neuronal responses to deep brain stimulation (DBS) at various targets, we have developed a protocol for ...

Time-dependent Increase in the Network Response to the Stimulation of Neuronal Cell Cultures on Micro-electrode Arrays

Micro-electrode arrays (MEAs) can be used to investigate drug toxicity, design paradigms for next-generation personalized medicine, and study network dynamics in neuronal cultures. In contrast with more traditional methods, such as patch-clamping, which can only record activity from a single cell, MEAs can record simultaneously from multiple sites in a network, without requiring the arduous task of placing each electrode individually. Moreover, numerous control and stimulation configurations can be easily applied within the same experimental setup, allowing for a broad range of dynamics to be explored. One of the key dynamics of interest in these in vitro studies has been the extent to which cultured networks display properties indicative of learning. Mouse neuronal cells cultured on MEAs display an increase in response following training induced by electrical stimulation. This protocol demonstrates how to culture neuronal cells on MEAs; successfully record from over 95% of the plated dishes; establish a protocol to train the networks to respond to patterns of stimulation; and sort, plot, and interpret the results from such experiments. The use of a proprietary system for stimulating and recording neuronal cultures is demonstrated. Software packages are also used to sort neuronal units. A custom-designed graphical user interface is used to visualize post-stimulus time histograms, inter-burst intervals, and burst duration, as well as to compare the cellular response to stimulation before and after a training protocol. Finally, representative results and future directions of this research effort are discussed.

Mouse neuronal cells cultured on multi-electrode arrays display an increase in response following electrical stimulation. This protocol demonstrates how to culture neurons, how to record activity, and how to establish a protocol to train the networks to respond to patterns of stimulation.

时间依赖性增加对微电极阵列刺激神经元细胞培养物的网络响应

Micro-electrode arrays (MEAs) can be used to investigate drug toxicity, design paradigms for next-generation personalized medicine, and study network ...

调查的意识到，不知道用功能磁共振成像的恐惧记忆的神经机制

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