这一材料是根据美国空军根据协议号 FA8650-16-2-6702 的研究而发起的。所表达的意见是作者的观点, 不反映国防部及其组成部分的官方观点或政策。美国政府有权为政府目的复制和分发重印, 尽管有任何版权批注。本研究所使用的受试者自愿、完全知情同意, 按 32 CFR 219 和多迪 3216.02_AFI 40-402 的要求获得。

以下 fMRI 测试协议符合莱特州立大学机构审查委员会提供的指导原则. 
 1. 控制组 <ol> <li> 仔细考虑并确定控制组 先验 。设计控制组允许对假设 (es) 进行评估, 并考虑由反馈所创建的其他因素 (如实践或期望) 的影响, 显示 44 . </li>
</ol> 2。硬件安装程序 <ol> <li> 在参与者进入 MRI 室之前准备所有硬件, 使用与传统 fMRI 相同的过程. </li>
	<li> 将 MR 兼容显示和响应设备系统连接到刺激计算机 (PC). </li>
	<li> 通过 MRI 孔或周围的 mr 兼容响应设备和耳机路由缆线. </li>
	<li> 将 TR 触发器输出从 MRI 连接到刺激 PC. 
	注意: 在某些设置中, 这可能连接到 MR 兼容的响应设备硬件, 然后连接到刺激计算机。这是同步刺激和数据采集的当务之急. </li>
	<li> 定位 MR 兼容显示器, 使其能够通过贴在磁头线圈上的镜子对参与者可见. </li>
</ol> 3。参与者定位 注意: 参与者应以与传统的 fMRI 相同的方式, 以类似的方式定位到扫描仪表上. 
<ol> <li> 使参与者在扫描仪表上的仰卧位置躺下。要求他们保持头部内的线圈. </li>
	<li> 将耳机放在参与者和 #39 的头部, 并确保耳朵被覆盖。如果需要额外的听力保护, 在定位耳机之前插入耳塞. </li>
	<li> 将垫子放在参与者和 #39 的膝盖下, 以增加舒适度. </li>
	<li> 将磁头线圈的上半身锁定到位. </li>
	<li> 将镜像粘贴到磁头线圈上. </li>
	<li> 将响应设备放置在参与者和 #39 的手中. </li>
	<li> 地标 nasion 相对于扫描仪的参与者和 #39 的位置. </li>
	<li> 将地标位置移动到 MRI 孔的中心. </li>
	<li> 确认参与者可以使用镜像查看整个显示。要求参与者根据需要调整镜像. </li>
</ol> 4。本地化目标区域 <ol> <li> 执行和 #34; 功能和 #34; 定位。使用功能本地化的大脑活动来定义感兴趣的目标区域 (ROI) 11 . 
	注意: 这一运行是以类似的方式执行传统的 fMRI。但是, 目标 ROI 也可以使用单独的解剖或标准化的地图集来定义, 以消除执行功能定位器的需要。
	<ol> <li> 向参与者提供脚本和/或可视任务说明. 
		注意: 这些说明应简洁, 但包含足够的信息, 使参与者能够成功执行在功能定位过程中执行的任务。在这里, 指示通知参与者一个点将在屏幕上, 他们可能会听到在耳机的声音。他们的目标是放松和关注点. </li>
		<li> 开始同步管理声音刺激 (, 如 双边连续白噪声 29 ) 和通过推 #34 进行数据获取; 扫描和 #34; MR 扫描仪上的按钮. 
		注意: 这是通过编程的表现的刺激使用 TR 触发器从 fMRI 获得。TR 触发器是通过 fMRI 协议控制的, 但是这可能会受到 MRI 制造商和安装的软件包的影响。任何视觉, 触觉, 和/或听觉刺激可以提供执行其他任务和/或目标的其他地区。
		<ol> <li> 将任务刺激 (白噪声) 与匹配的控制刺激 (无噪音) 的传递替换为阻塞模式。使用控制刺激激活在任务刺激中激活的不需要的网络/系统. 
			注意: 这种交替发生的同步刺激的 fMRI 获得和监测 TR 脉冲. </li>
			<li> 利用梯度回波成像脉冲序列采集全脑回波平面图像; 脉冲序列的示例参数包括在相位和频率方向上的 64 x 64 元素的采集矩阵, 41 片平行于前合-后合平面, 3.75 x 3.75 x 3 mm 3 体素大小, 0.5 mm 切片间隙, 启用脂肪抑制, TR/TE = 2000/20 ms, 和一个翻转角度 = 90 & #176;. </li>
		</ol> </li> </ol> </li> <li> 使用多变量统计信息在功能定位器期间收集的 fMRI 数据计算激活映射。
	注意: 以下步骤是对传统 fMRI 进行的处理的变体。已删除或简化了某些步骤以减少处理时间。
	<ol> <li> 使用从标准预处理技术创建的自定义软件 12 、 45 对数据进行预处理。
		<ol> <li> 使用高斯低通内核执行3D 空间过滤 (全角 half-maximum 4.5 mm). </li>
			<li> 通过三线性插值将每个卷的质心与功能定位器的第一个音量对齐, 从而校正平移运动. </li>
			<li> 使用与 #963 的高斯低通内核执行时间过滤; = 3 s. </li>
		</ol> </li> <li> 创建一个模型来预测任务的神经响应; 这是以与传统 fMRI 相同的方式执行的。
		<ol> <li> 创建一个心理模型, 它描述每个时间点 46 的活动和 rest 状态。这个模型的时间点在任务期间与价值 #39; 1 和 #39; 和控制与 #39; 0 和 #39;. </li>
			<li> Convolve 具有预定义的血流动力学响应函数 (HRF) 46 的心理模型, 以预测该任务的 fMRI (神经) 响应. </li>
		</ol> </li> <li> 使用一般线性模型 (GLM) 将每个体素中的 fMRI 数据与神经模型的时间函数相匹配。这将导致 和 #946; 参数映射, 它将使用标准统计转换转换为 t 或 z 统计映射 (激活映射). </li>
	</ol> </li> <li> 使用覆盖在平均功能磁共振成像图像上的激活映射来确定将在其中派生后续神经的反馈信号的区域. 
	注意: 这是使用自定义软件执行的。要删除全局和不的更改, 还可以定义第二个 ROI。
	<ol> <li> 使用鼠标滑块滚轮或切片滑动条在切片中查找在 f 中可见的解剖标记。MRI 数据, 如侧脑室额角的下表面 12 . </li>
		<li> 使用阈值滑动条来阈值激活映射, 以显示目标区域中功能定位器中最强劲的体激活。
		<ol> <li> 通过选择阈值 先验 或手动调整阈值来执行此操作. </li>
		</ol> </li> <li> 使用鼠标左键选择单个体, 并在选定阈值的上方激活, 并在目标区域内添加到 ROI 中. 
		注意: 可以从一个或多个切片中选择体. </li>
	</ol> </li> </ol> 5. fMRI 测试 <ol> <li> 使用具有交替任务和控制条件的车厢模型进行神经运行。
	<ol> <li> 实现一个任务条件, 其中参与者提高或降低目标区域的活动, 而控制方向对实现所需的结果至关重要. 
		注: 例如, 大脑的许多区域在耳鸣患者中是活跃的, 因此, 减少活动可能会鼓励正常的神经模式. </li>
		<li> 将任务条件替换为一个控制条件, 参与者通过放松和清除他们的思想来返回活动休息. </li>
		<li> 为参与者提供一个在两种情况下都要使用的正念任务的脚本示例, 作为启动对所需状态的大脑活动的辅助工具。指导参与者执行正念任务, 推动大脑活动朝向所需状态。
		<ol> <li> 在耳鸣示例中, 指示参与者将注意力从听觉系统转移到其他感官系统, 以减少听觉活动. </li>
		</ol> </li> <li> 比较基准计算 注意: 由于在每次运行之前都对 MRI 硬件组件进行了调整, 因此基线用于在向参与者提交反馈之前对数据进行规范化。基线平均值是为目标区域确定的, 使用在每个 fMRI 测试开始时获得的一个或多个卷的平均值, 运行 12 , 47 。
		<ol> <li> 指示参与者在扫描开始时显示的倒计时中放松. #160; </li> </ol> </li>
	</ol> </li> <li> 通过按 #34; 扫描和 #34; MRI 扫描仪上的按钮, 开始同步刺激演示和数据获取。使用梯度召回回波 MRI 脉冲序列收集回声平面图像, 其方法与步骤4.1.2 中的功能定位器所规定的相同. </li>
	<li> 获取基线卷。
	<ol> <li> 直观显示倒计时计时器和空白反馈显示器. </li>
		<li> 使用自定义软件在获取过程中处理数据。
		<ol> <li> 使用高斯低通内核执行3D 空间过滤 (全角 half-maximum 4.5 mm). </li>
			<li> 使用每个卷的质心对平移运动进行校正; 每个卷都通过三线插值注册到功能定位器的第一个卷. </li>
			<li> 在时间和空间上计算来自目标 ROI 的平均信号. </li>
			<li> 对每个卷中目标 ROI 中的所有体的信号进行求和. </li>
			<li> 通过将总和除以 roi 中的体数, 为每个卷创建 roi 平均值. </li>
			<li> 平均来自基线卷的总和. </li>
		</ol> </li> </ol> </li> <li> 获取神经卷 <ol> <li> 在使用自定义软件获取过程中对数据进行预处理。
		<ol> <li> 使用高斯低通内核执行3D 空间过滤 (全角 half-maximum 4.5 mm). </li>
			<li> 通过三线性插值将每个卷的质心与功能定位器的第一个音量对齐, 从而校正平移运动. </li>
		</ol> </li> <li> 计算反馈信号。在 fMRI 测试中, 每个获得的体积都有一个反馈信号。这是向参与者提供的帮助学习意志控制的信息。
		<ol> <li> 平均从目标 ROI 中所有体的 fMRI 信号创建单个值. </li>
			<li> 计算当前 roi 平均值和 roi 基线平均值之间的百分比变化。(可选) 此信号可以通过依赖于参与者和 #39 性能的因素进行缩放. </li>
			<li> 计算反馈信号由世俗地过滤 (高斯低通核与 3 s 的斯格码组成仅过去组分) 当前百分比变动与反馈信号从早先神经容量. </li>
		</ol> </li> <li> 显示反馈信号。
		<ol> <li> 通过温度计样式的条形图显示当前的反馈信号, 其中条形的高度与反馈值成正比 18 、 19 、 21 , 34 . </li>
			<li> 在反馈显示上为参与者覆盖说明. 
			注意: 这些说明很简单, 应该指导参与者放松, 或提高或降低活动 ( 即 温度计栏). </li>
		</ol> </li> <li> 可选择提供额外的刺激。额外的视觉, 听觉, 或触觉刺激可以同时提供反馈. </li>
	</ol> </li> </ol> 6。评估自我调节目标 ROI 的能力. 
 注意: 在神经完成后, 需要对每个培训的目标区域自我的能力进行量化. 
<ol> <li> 分析反馈信号 12 中的 intra-subject 更改。
	<ol> <li> 创建一个表示神经的 rest 和任务条件的心理模型. 
		注意: 此模型与预定义的 HRF 积, 以生成神经模型。该进程与为功能定位程序所描述的过程相同. </li>
		<li> 使用 GLM 将反馈信号时间序列与神经模型相匹配。这将导致 and #946; 参数转换为 t- 或 z 统计代表的能力, 自我. </li>
	</ol> </li> <li> 执行学科比较. 
	注意: 可以通过适当的统计分析 ( 例如, #160; 配对 t 测试或 ANOVAs), 将自我调节性能的统计数据代表在不同的运行和组之间进行比较。这些测试评估在训练和组之间自我目标区域的能力的变化, 并可用于评估研究和 #39 的假说. </li>
</ol>

作者没有什么可透露的。

本文讨论的 fMRI 测试协议可以适应任何区域的大脑, 并讨论了一个单变量, 基于的方法, 以神经。这可以通过编程其他功能的本地化任务来激活其他区域来实现。通过将这些任务集成到自定义神经软件中, 我们开发了一个非常简单的过程。但是, 有一个限制: 目标区域必须在功能上定义。此时, 我们的团队开发的软件不会在功能和解剖图像之间进行任何注册。因此, 其他 ROI 选择方法 (如 atlas-based roi) 此时无法实现。此外 , 对刺激和神经的参数 (例如 ,块持续时间、块数和包括 TR 在内的成像参数 ) 可以很容易作者操纵。此外, 在没有神经的情况下, 传输运行以评估自我目标 ROI 的能力是可以实现的。我们开发的软件不提供神经利用多元模式35,48或大脑区域之间的连接49。
FMRI 测试比其他形式的神经提供了显著的优势, 但也有其局限性。功能磁共振成像测试的主要优势是空间分辨率优于所有其他形式的测试, 如脑电图 (EEG) 为基础的神经。增强的空间分辨率使特定的大脑结构/功能在整个大脑中成为目标50。目前, 这是无法实现的其他疗法, 如药物治疗, 这是系统性的。然而, fMRI 测试的主要缺点是时间延迟。不仅采样率比脑电图慢得多 (高达3的数量级减慢), 与 fMRI 信号相关的血流动力学滞后进一步增加了这一延迟。尽管如此, 有压倒性的证据表明, 参与者可以克服这一延迟, 并在实践中学会控制大脑活动 (例如查看查看苏尔寿et al.11和 Scharnowski et al.50)。
fMRI 测试的普及程度正在上升, 但仍处于起步阶段。由于这一原因, 尚未采取共同的做法。所描述的协议细节方法是科学地被接受的。例如, 多种形式的反馈显示已在各种研究中使用, 包括温度计样式的条形图18,19,21,34。此外, 一个反馈信号, 作为百分比信号的变化与基线计算从目标区域也已广泛实施12,19,21,25,30,51,52。
控制大脑中的塑料效果提供了一种创新的治疗技术来治疗神经紊乱或大脑异常活动, 例如与上述的耳鸣相关的脑损伤。虽然将调节转化为行为效应的确切机制尚不得而知, 但 fMRI 测试已经与11相关。通过学习过程, 当一个人在任务相关的大脑网络中积极调节大脑活动时, 行为会得到加强。这种强化导致了神经机制的参与, 使网络更有效率地执行。这与其他测试技术 (如脑神经) 不谋而合, 那里的个人被训练来控制从头皮的本地区域测量的电信号频带53,54,55.其他人已经表明从突触可塑性, 导致提高突触效率12。还有一个假设建议, 细胞机制的学习可能涉及电压依赖性膜电导的变化, 这是表现为神经兴奋性的变化13。在任何情况下, 似乎 fMRI 测试导致细胞水平的变化, 并且个体可能学会对这些过程的一些控制。这种能力和这些变化可能是关键的学习和发展治疗脑损伤和神经系统紊乱。
fMRI 测试的一个重要方面是测量行为的改变。这是必要的许多假说预测行为的变化驱动的测试诱导神经变化。至少应在两个时间点收集这些评估: 在测试之前和之后。在耳鸣的情况下, 这些行为评估可能只包括主观问卷, 因为没有直接的措施, 耳鸣。对于其他神经系统疾病, 应进行文献复习, 以确定对所调查的特定假说的适当、合理和有记录的评估。一些假说需要在额外的时间点测量, 比如那些探索 fMRI 测试近, 短期和长期影响的研究。有些评估可能需要在测试之前进行培训, 以减少学习效果。其他假说甚至可能需要神经系统测试, 比如那些对脑代谢物、脑灌注或功能网络有兴趣的人。
fMRI 测试程序有两个关键阶段。第一个是确定一个大脑区域以神经为目标。在进行任何程序之前, 应进行彻底的文献复习, 以研究神经通路和与神经系统紊乱或脑损伤相关的重要结构/功能。因此, 关键的结构/功能应被仔细选择作为神经的目标。接下来, 应进行另一次文献复习, 以检查与此结构/功能相关的任务。此任务可能与无序相关联, 但应确认任务在指定的填充中激活所需的区域。在神经过程中, 这一目标区域将在第一届会议或每届会议上逐个选定。因此, 和 intra-subject 变异性可能是导致不可预知结果的重要因素。创建一个协议来选择目标区域并进行适当的人员培训是至关重要的。有两种定义目标 ROI 的方法: 解剖和功能。解剖定义利用结构 MRI 扫描, 严格定义目标区域从解剖,并可能使用标准的地图集。功能图像注册到结构图像, 目标区域转换为功能空间21,26。在函数方法中, 目标区域是通过执行功能本地化11、12、24、29、44所产生的激活映射来选择的。本文讨论了这种方法。
fMRI 测试的第二个关键阶段是控制组选择。对照组在确定 fMRI 测试的作用时至关重要, 应仔细考虑控制组的选择。以前的研究使用了广泛的控制。控制组的一个常见过程是在存在虚假反馈的情况下尝试意志控制。此反馈可以从实验组的参与者21,44提供, 从一个不参与所需进程的区域中获得, 参与者17,33,44, 或倒置52。其他研究使用了试图意志控制的控制组, 但未提供神经12、21、44、56。
先前的一项研究表明, 当受试者试图控制假反馈时, 在双侧脑岛、前扣带、辅助马达、背和侧前额区的活化作用与被动观察反馈显示57。这些发现牵连到一个广泛的额壁和 cingulo-盖网络是激活时, 有意图控制大脑活动。此外, 这些发现表明, 传统的控制组使用的测试实验将使用神经相关的一致性与认知控制, 即使在存在的虚假反馈。一个单独的荟萃分析揭示了前岛和基底神经节的活动, 这两个区域都是认知控制和其他高级认知功能的组成部分, 是试图意志控制58的关键因素。meta 分析的结果证实了先前的发现57。在一起, 这一证据表明, 这是至关重要的描绘成功的意志控制的效果和那些与尝试自我调节。因此, 包括不尝试自我规管的控制小组可能是重要的。
然而, 以前的研究中, 控制组收到的假功能磁共振成像信号显示, 目标 ROI 活动的差异, 从那些收到真正的反馈15,16,17,18,20,21,25,26,28,33,34,44, 暗示不包含反馈的培训策略在调整目标区域时并不有效。此外, 控制组收到相同的指示和相同的训练期间, 但没有收到反馈的大脑活动水平, 没有表现出类似的行为结果的实验组谁给神经12,18,21,32,44,59。这些结果表明, 经验的影响是由于 fMRI 测试诱导的学习, 而不是其他学习或非特异性的变化。因此, 必须制定具体的培训方案, 针对特定的神经系统, 以获得预期的效果。与各种对照组的研究结果表明, 行为训练, 实践, 感官反馈和生物反馈单独不产生等效的行为效果, 那些谁接受 fMRI 测试44。

神经紊乱对受影响的个人、他们的家庭和社会造成了严重的障碍。治疗神经紊乱可能是不存在的或有疑问的功效, 往往只是针对症状的紊乱。这种情况就是耳鸣--幻像的声音--没有得到美国食品和药物管理局 (FDA) 批准的治疗。耳鸣可以对一个人的生活产生深远的影响, 通过降低注意力或改变对实际声音的感知来干扰日常任务。此外, 受耳鸣影响的个人也可能经历疲劳、压力、睡眠问题、记忆力问题、抑郁、焦虑和烦躁不安1。确实存在的治疗方法, 如抗抑郁药和焦虑药物, 只会帮助管理相关的症状, 而不能治疗潜在的病因。这为这些疾病的创新治疗创造了一个关键的缺口。
在采集技术、计算能力和算法方面的改进已经彻底改变了功能性磁共振成像 (fMRI) 数据的测量和处理速度。这使得 real-time 功能磁共振成像的问世, 数据可以处理, 因为它是收集。早期应用的 real-time 功能磁共振成像是有限的2, 主要是由于无法快速完成的预处理步骤, 典型的离线分析, 如运动矫正。计算技术和算法的改进现在提高了 real-time fMRI3的速度、灵敏度和通用性, 允许在 real-time 中应用类似的离线预处理。这些发展导致了4主要应用领域 real-time fMRI: 术中手术指导4, 脑-计算机接口5,6, 适应当前脑状态的刺激7,神经培训8。
测试, 虽然不是最初的重点 real-time fMRI, 是一个不断增长的研究领域, 个人学习调节大脑活动特定通过实施的心理策略 (即想象的任务)。测试是一种操作性条件反射的形式9, 它已经被证明增加神经元的射击率和神经元活动在猴子10。此外, fMRI 测试已经与钉定时相关的可塑性, 这是神经变化发生在联想学习11。进一步的暗示表明, fMRI 测试通过长期的增强作用诱导了可塑性, 从而提高了突触效率12。另一个假设意味着技能学习的细胞机制, 如意志控制大脑活动, 并可能涉及电压依赖性的细胞膜电导的变化-表现为神经兴奋性的变化13。在任何情况下, 似乎 fMRI 测试影响大脑的神经水平。这些理论为使用 fMRI 测试治疗神经系统疾病提供了有力的理由。
fmri 测试, 不同于传统的 fmri, 提供了一个机会来研究大脑活动和行为之间的关系11,14。最近, 在涉及 fMRI 测试的研究中, 与前10年 (n = 16)11相比, 2011年发表的文章的数量几乎是 2012 (n = 30) 的两倍。第一个 fMRI 测试研究是由 Weiskopf 和同事在 2003年8进行的。本研究成功地展示了在前扣带皮层 (ACC) 中使用一个参与者的在线反馈和 fMRI 信号的自我调节的可行性。反馈显示延迟约两秒, 超过一个数量级比以前的研究快得多。第一个完整的研究是在2004年进行的, 6 参与者学会了控制 somatomotor 皮层的活动15。FMRI 测试在同一天完成了3次会议。通过在 single-subject 和组水平的训练过程中观察到在 somatomotor 皮层中对目标区域的空间选择性增加的活动。这个效果是没有观察到的控制组收到真实的功能磁共振成像信息从一个背景区域 (不相关的任务正在执行) 在前面的运行。研究人员已经表明, 人类可以学习意志控制的 fMRI 信号测量从许多大脑区域, 包括 ACC16, 杏仁核17, 前脑岛18,19, 听觉和注意相关网络20, 双边侧前额叶皮层21, 侧前额叶皮层12,22,23, 运动神经皮层24, 25,26,27,28, 主要听觉皮层29,30, 与情绪网络区域相关的区域31,32, 右下额上回33, 和视觉皮层34,35。
许多神经系统疾病的潜在机制是未知的。在耳鸣的例子中, 在大多数情况下没有明显的幻像源36,37,38。尽管如此, 证据表明, 一个中央机制可能是负责的耳鸣感知在某些个人, 证明了缺乏症状解决后, 完整的听觉神经解剖39。与耳鸣相关的多动症已在初级听觉皮层40、41、42中找到。进一步的证据表明, 耳鸣的影响进一步扩大到参与处理情感和注意状态的领域43。基于这些异常, fMRI 测试范式可以发展, 以诱导和控制神经机制, 鼓励正常的神经模式。

<table border="0" cellpadding="0" cellspacing="0" width="639">
	<tbody>
		<tr height="20">
			<td height="20" style="height:20px;width:187px;">3T MRI</td>
			<td style="width:136px;">GE Medical</td>
			<td style="width:93px;">750W Discovery</td>
			<td style="width:223px;">Data Acquisition Hardware</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">MR-Compatible Display System</td>
			<td>InVivo</td>
			<td>SensaVue</td>
			<td>Visual Stimuli Hardware</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">MR-Compatible Auditory System</td>
			<td>Resonance Technologies</td>
			<td>CinemaVision</td>
			<td>Auditory Stimuli Hardware</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Experimental Stimulus Software</td>
			<td>Neurobehavioral Systems</td>
			<td>Presentation</td>
			<td>Software to Control Stimuli Presentation</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Experimental Processing Software</td>
			<td>Mathworks</td>
			<td>MATLAB</td>
			<td>Software to Process Data</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Data Processing Software</td>
			<td>Microsoft</td>
			<td>Visual Studio C++</td>
			<td>Software to Process Data</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Response Pads</td>
			<td>Cedrus Corporation</td>
			<td>Lumina</td>
			<td>Hardware to Receive Participant Input</td>
		</tr>
	</tbody>
</table>

a protocol for the administration of real-time fmri neurofeedback training

神经疾病的特点是在大脑中的细胞、分子和电路功能异常。新的方法诱导和控制神经过程和纠正异常功能, 甚至转移功能从受损的组织到生理健康的脑区, 持有潜力, 以显著改善整体健康。在目前的神经干预的发展, 神经训练 (测试) 从功能磁共振成像 (fMRI) 具有完全无创, 力度, 和空间定位到目标大脑的优势区域, 也没有已知的副作用。此外, 测试技术, 最初开发使用 fMRI, 往往可以转化为演习, 可以在扫描仪以外的情况下进行, 没有医疗专业人员或尖端医疗设备的援助。在 fmri 测试, fmri 信号是测量从大脑的特定区域, 处理, 并提交给参与者在 real-time。通过训练, 自我导向的心理处理技术, 即调节这一信号及其潜在的神经关联, 被开发。FMRI 测试已经被用来训练意志控制的广泛的大脑区域, 对几个不同的认知, 行为和马达系统的影响。此外, 功能磁共振成像测试已经表明, 在广泛的应用, 如治疗神经紊乱和增加基线人类的表现的承诺。在这篇文章中, 我们提出了一个功能磁共振成像测试协议, 在我们的机构发展, 以调节健康和异常的大脑机能, 以及使用的方法, 以认知和听觉区域的大脑的例子。

我们的团队已经证明了在18参与者的队列中从 fMRI 测试学到的左侧前额皮质 (DLPFC) 的控制显著增加。在意志控制12的定量值上, 对受试者进行了 one-way 方差分析。这项分析显示, 左 DLPFC 的控制显著增加了 5 x 6 min:24 运行的神经在 14 d (图 1中分别进行了五个独立的会话;F(468) = 2.216, p = 0.038, 度假设, 单尾)。在测试之前和之后执行的复杂多任务测试的性能变化, 与未接受神经使用 2 x 2 混合模型 ANOVAs 的组进行比较。经过 Bonferroni 纠正后的比较发现, 复杂的多任务测试的性能显著提高, 没有接受额外的培训 (p & #60; 0.005, 一尾), 而且这一增幅明显大于对照组执行类似的培训, 但没有提供额外的援助神经 (p & #60; 0.03, 度假设, 一尾)12。尽管实验组在训练中获得了对左 DLPFC 的控制, 但没有观察到一个高原。这意味着不需要最大程度的控制来产生行为结果, 并且在进一步培训12时可能会有更大的影响。此外 , 我们的团队发现功能磁共振成像测试结合n- 背部练习创建的大脑活动的焦点变化 , 限制在目标区域 , 不影响工作内存网络的向上或向组件 (图 2)22。
关于耳鸣, 一项先前的研究已经调查 fMRI 测试作为一种可能的治疗29。在这项研究中, 4 x 4 分钟的神经的运行完成了一个单一的培训课程。在单一功能磁共振成像测试会议之前和之后进行了耳鸣的行为评估。成功的意志下调的听觉皮层, 并导致显著减少听觉活化。本研究表明 fMRI 测试在治疗耳鸣方面的前景, 然而, 仅对六参与者进行了研究, 对照组未被用于比较。此外, 不进行统计分析, 包括行为数据。在这项研究的基础上扩大可能会发现耳鸣患者的新治疗机会。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/55543/55543fig1.jpg"> 
图 1: 增加对左 DLPFC 的控制.每个神经运行的平均左 DLPFC 控制 (在不同的天执行) 由淡绿色圆圈表示。线性回归分析显示, 在训练中的控制显著增加 (暗绿色线; β = 1.078, p & #60; 0.033)。误差线代表 1 SEM. 舍伍德的未修改工作et al.12, 在 "创作共享" 归属许可下重印。<a href="http://ecsource.jove.com/files/ftp_upload/55543/55543fig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/55543/55543fig2.jpg"> 
图 2: 学习左 DLPFC 控制的本地化效果.(A) 从n回功能定位仪中选择的 fMRI 测试的体素包含概率。在测试靶区中最常见的是浅蓝色体, 暗蓝色体的频率较低, 不包括透明体。(B) 基于培训课程 (红-黄) 的主要效果的方差分析结果。这一结果显示出一个大的重叠与左 DLPFC roi 目标为测试。轴向切片在放射学大会在坐标 z = 22, 26, 30, 34 和38毫米 (从左到右) 显示。舍伍德的未修改工作et al.22, 在 "创作共享" 归属许可下重印。<a href="http://ecsource.jove.com/files/ftp_upload/55543/55543fig2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>

Watch this Scientific Journal Video about A Protocol for the Administration of Real-Time fMRI Neurofeedback Training at JoVE.com

A Protocol for the Administration of Real-Time fMRI Neurofeedback Training

诱导和/或控制神经可塑性的能力可能是未来治疗神经系统紊乱和脑损伤恢复的关键。本文提出了神经训练与功能磁共振成像技术在人脑功能调节中的应用。

实时 fMRI 神经训练管理协议

Neurologic disorders are characterized by abnormal cellular-, molecular-, and circuit-level functions in the brain. New methods to induce and control ...

a-protocol-for-administration-real-time-fmri-neurofeedback

Research

JoVE Journal

Neuroscience

10.9K 次观看.  Wright State University. 诱导和/或控制神经可塑性的能力可能是未来治疗神经系统紊乱和脑损伤恢复的关键。本文提出了神经训练与功能磁共振成像技术在人脑功能调节中的应用。

视频: A Protocol for the Administration of Real-Time fMRI Neurofeedback Training

Combining Transcranial Magnetic Stimulation and fMRI to Examine the Default Mode Network

The default mode network is a group of brain regions that are active when an individual is not focused on the outside world and the brain is at "wakeful rest."1,2,3 It is thought the default mode network corresponds to self-referential or "internal mentation".2,3
It has been hypothesized that, in humans, activity within the default mode network is correlated with certain pathologies (for instance, hyper-activation has been linked to schizophrenia 4,5,6 and autism spectrum disorders 7 whilst hypo-activation of the network has been linked to Alzheimer's and other neurodegenerative diseases 8). As such, noninvasive modulation of this network may represent a potential therapeutic intervention for a number of neurological and psychiatric pathologies linked to abnormal network activation. One possible tool to effect this modulation is Transcranial Magnetic Stimulation: a non-invasive neurostimulatory and neuromodulatory technique that can transiently or lastingly modulate cortical excitability (either increasing or decreasing it) via the application of localized magnetic field pulses.9
In order to explore the default mode network's propensity towards and tolerance of modulation, we will be combining TMS (to the left inferior parietal lobe) with functional magnetic resonance imaging (fMRI). Through this article, we will examine the protocol and considerations necessary to successfully combine these two neuroscientific tools.

In this article, we examine the methodology and considerations relevant to the combination of TMS and fMRI to examine the effects of brain stimulation on the default network.

结合经颅磁刺激和功能磁共振成像检查默认模式网络

The default mode network is a group of brain regions that are active when an individual is not focused on the outside world and the brain is at "wakeful ...

科研

神经科学

Organotypic Slice Culture of GFP-expressing Mouse Embryos for Real-time Imaging of Peripheral Nerve Outgrowth

For many purposes, the cultivation of mouse embryos ex vivo as organotypic slices is desirable. For example, we employ a transgenic mouse line (tauGFP) in which the enhanced version of the green fluorescent protein (EGFP) is exclusively expressed in all neurons of the developing central and peripheral nervous system1, allowing the possibility to both film the innervation of the forelimb and to manipulate this process with pharmacological and genetic techniques2. The most critical parameter in the successful cultivation of such slice cultures is the method by which the slices are prepared. After extensive testing of a variety of methods, we have found that a vibratome is the best possible device to slice the embryos such that they routinely result in a culture that demonstrates viability over a period of several days, and most importantly, develops in an age-specific manner. For mid-gestation embryos, this includes the normal outgrowth of spinal nerves from the spinal cord and the dorsal root ganglia to their targets in the periphery and the proper determination of skeletal and muscle tissue. 
In this work, we present a method for processing whole embryos of embryonic day (E) E10 to E12 into 300 - 400 micrometer slices for cultivation in a standard tissue culture incubator, which can be studied for up to two days after slice preparation. Critical for the success of this approach is the use of a vibratome to slice each agarose-embedded embryo. This is followed by the cultivation of the slices upon Millicell culture membrane inserts placed upon a small volume of medium, resulting in an interface culture technique. One litter with an average of 7 embryos routinely produces at least 14 slices (2-3 slices of the forelimb region per embryo), which varies slightly due to the age of the embryos as well as to the thickness of the slices. About 80% of the cultured slices show nerve outgrowth, which can be measured througout the culturing period2. Representative results using the tauGFP mouse line are demonstrated.

We present a method to prepare organotypic slices of mid-gestation mouse embryos for the cultivation and time-lapse imaging of peripheral nerve outgrowth.

器官型切片文化绿色荧光蛋白表达的实时成像周围神经生长的小鼠胚胎

For many purposes, the cultivation of mouse embryos ex vivo as organotypic slices is desirable. For example, we employ a transgenic mouse line (tauGFP) in ...

The NeuroStar TMS Device: Conducting the FDA Approved Protocol for Treatment of Depression

The Neuronetics NeuroStar Transcranial Magnetic Stimulation (TMS) System is a class II medical device that produces brief duration, pulsed magnetic fields. These rapidly alternating fields induce electrical currents within localized, targeted regions of the cortex which are associated with various physiological and functional brain changes.1,2,3
In 2007, O'Reardon et al., utilizing the NeuroStar device, published the results of an industry-sponsored, multisite, randomized, sham-stimulation controlled clinical trial in which 301 patients with major depression, who had previously failed to respond to at least one adequate antidepressant treatment trial, underwent either active or sham TMS over the left dorsolateral prefrontal cortex (DLPFC). The patients, who were medication-free at the time of the study, received TMS five times per week over 4-6 weeks.4
The results demonstrated that a sub-population of patients (those who were relatively less resistant to medication, having failed not more than two good pharmacologic trials) showed a statistically significant improvement on the Montgomery-Asberg Depression Scale (MADRS), the Hamilton Depression Rating Scale (HAMD), and various other outcome measures. In October 2008, supported by these and other similar results5,6,7, Neuronetics obtained the first and only Food and Drug Administration (FDA) approval for the clinical treatment of a specific form of medication-refractory depression using a TMS Therapy device (FDA approval K061053).
In this paper, we will explore the specified FDA approved NeuroStar depression treatment protocol (to be administered only under prescription and by a licensed medical profession in either an in- or outpatient setting).

In this article, we examine the methodology and considerations relevant to the FDA approved depression treatment protocol using the Neuronetics NeuroStar TMS device.

NeuroStar TMS的设备进行美国FDA批准用于治疗抑郁症的议定书"

The Neuronetics NeuroStar Transcranial Magnetic Stimulation (TMS) System is a class II medical device that produces brief duration, pulsed magnetic ...

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 ...

Recording Human Electrocorticographic (ECoG) Signals for Neuroscientific Research and Real-time Functional Cortical Mapping

Neuroimaging studies of human cognitive, sensory, and motor processes are usually based on noninvasive techniques such as electroencephalography (EEG), magnetoencephalography or functional magnetic-resonance imaging. These techniques have either inherently low temporal or low spatial resolution, and suffer from low signal-to-noise ratio and/or poor high-frequency sensitivity. Thus, they are suboptimal for exploring the short-lived spatio-temporal dynamics of many of the underlying brain processes. In contrast, the invasive technique of electrocorticography (ECoG) provides brain signals that have an exceptionally high signal-to-noise ratio, less susceptibility to artifacts than EEG, and a high spatial and temporal resolution (i.e., &lt;1 cm/&lt;1 millisecond, respectively). ECoG involves measurement of electrical brain signals using electrodes that are implanted subdurally on the surface of the brain. Recent studies have shown that ECoG amplitudes in certain frequency bands carry substantial information about task-related activity, such as motor execution and planning1, auditory processing2 and visual-spatial attention3. Most of this information is captured in the high gamma range (around 70-110 Hz). Thus, gamma activity has been proposed as a robust and general indicator of local cortical function1-5. ECoG can also reveal functional connectivity and resolve finer task-related spatial-temporal dynamics, thereby advancing our understanding of large-scale cortical processes. It has especially proven useful for advancing brain-computer interfacing (BCI) technology for decoding a user's intentions to enhance or improve communication6 and control7. Nevertheless, human ECoG data are often hard to obtain because of the risks and limitations of the invasive procedures involved, and the need to record within the constraints of clinical settings. Still, clinical monitoring to localize epileptic foci offers a unique and valuable opportunity to collect human ECoG data. We describe our methods for collecting recording ECoG, and demonstrate how to use these signals for important real-time applications such as clinical mapping and brain-computer interfacing. Our example uses the BCI2000 software platform8,9 and the SIGFRIED10 method, an application for real-time mapping of brain functions. This procedure yields information that clinicians can subsequently use to guide the complex and laborious process of functional mapping by electrical stimulation.
Prerequisites and Planning:
Patients with drug-resistant partial epilepsy may be candidates for resective surgery of an epileptic focus to minimize the frequency of seizures. Prior to resection, the patients undergo monitoring using subdural electrodes for two purposes: first, to localize the epileptic focus, and second, to identify nearby critical brain areas (i.e., eloquent cortex) where resection could result in long-term functional deficits. To implant electrodes, a craniotomy is performed to open the skull. Then, electrode grids and/or strips are placed on the cortex, usually beneath the dura. A typical grid has a set of 8 x 8 platinum-iridium electrodes of 4 mm diameter (2.3 mm exposed surface) embedded in silicon with an inter-electrode distance of 1cm. A strip typically contains 4 or 6 such electrodes in a single line. The locations for these grids/strips are planned by a team of neurologists and neurosurgeons, and are based on previous EEG monitoring, on a structural MRI of the patient's brain, and on relevant factors of the patient's history. Continuous recording over a period of 5-12 days serves to localize epileptic foci, and electrical stimulation via the implanted electrodes allows clinicians to map eloquent cortex. At the end of the monitoring period, explantation of the electrodes and therapeutic resection are performed together in one procedure.
In addition to its primary clinical purpose, invasive monitoring also provides a unique opportunity to acquire human ECoG data for neuroscientific research. The decision to include a prospective patient in the research is based on the planned location of their electrodes, on the patient's performance scores on neuropsychological assessments, and on their informed consent, which is predicated on their understanding that participation in research is optional and is not related to their treatment. As with all research involving human subjects, the research protocol must be approved by the hospital's institutional review board. The decision to perform individual experimental tasks is made day-by-day, and is contingent on the patient's endurance and willingness to participate. Some or all of the experiments may be prevented by problems with the clinical state of the patient, such as post-operative facial swelling, temporary aphasia, frequent seizures, post-ictal fatigue and confusion, and more general pain or discomfort.
At the Epilepsy Monitoring Unit at Albany Medical Center in Albany, New York, clinical monitoring is implemented around the clock using a 192-channel Nihon-Kohden Neurofax monitoring system. Research recordings are made in collaboration with the Wadsworth Center of the New York State Department of Health in Albany. Signals from the ECoG electrodes are fed simultaneously to the research and the clinical systems via splitter connectors. To ensure that the clinical and research systems do not interfere with each other, the two systems typically use separate grounds. In fact, an epidural strip of electrodes is sometimes implanted to provide a ground for the clinical system. Whether research or clinical recording system, the grounding electrode is chosen to be distant from the predicted epileptic focus and from cortical areas of interest for the research. Our research system consists of eight synchronized 16-channel g.USBamp amplifier/digitizer units (g.tec, Graz, Austria). These were chosen because they are safety-rated and FDA-approved for invasive recordings, they have a very low noise-floor in the high-frequency range in which the signals of interest are found, and they come with an SDK that allows them to be integrated with custom-written research software. In order to capture the high-gamma signal accurately, we acquire signals at 1200Hz sampling rate-considerably higher than that of the typical EEG experiment or that of many clinical monitoring systems. A built-in low-pass filter automatically prevents aliasing of signals higher than the digitizer can capture. The patient's eye gaze is tracked using a monitor with a built-in Tobii T-60 eye-tracking system (Tobii Tech., Stockholm, Sweden). Additional accessories such as joystick, bluetooth Wiimote (Nintendo Co.), data-glove (5th Dimension Technologies), keyboard, microphone, headphones, or video camera are connected depending on the requirements of the particular experiment.
Data collection, stimulus presentation, synchronization with the different input/output accessories, and real-time analysis and visualization are accomplished using our BCI2000 software8,9. BCI2000 is a freely available general-purpose software system for real-time biosignal data acquisition, processing and feedback. It includes an array of pre-built modules that can be flexibly configured for many different purposes, and that can be extended by researchers' own code in C++, MATLAB or Python. BCI2000 consists of four modules that communicate with each other via a network-capable protocol: a Source module that handles the acquisition of brain signals from one of 19 different hardware systems from different manufacturers; a Signal Processing module that extracts relevant ECoG features and translates them into output signals; an Application module that delivers stimuli and feedback to the subject; and the Operator module that provides a graphical interface to the investigator.
A number of different experiments may be conducted with any given patient. The priority of experiments will be determined by the location of the particular patient's electrodes. However, we usually begin our experimentation using the SIGFRIED (SIGnal modeling For Realtime Identification and Event Detection) mapping method, which detects and displays significant task-related activity in real time. The resulting functional map allows us to further tailor subsequent experimental protocols and may also prove as a useful starting point for traditional mapping by electrocortical stimulation (ECS).
Although ECS mapping remains the gold standard for predicting the clinical outcome of resection, the process of ECS mapping is time consuming and also has other problems, such as after-discharges or seizures. Thus, a passive functional mapping technique may prove valuable in providing an initial estimate of the locus of eloquent cortex, which may then be confirmed and refined by ECS. The results from our passive SIGFRIED mapping technique have been shown to exhibit substantial concurrence with the results derived using ECS mapping10.
The protocol described in this paper establishes a general methodology for gathering human ECoG data, before proceeding to illustrate how experiments can be initiated using the BCI2000 software platform. Finally, as a specific example, we describe how to perform passive functional mapping using the BCI2000-based SIGFRIED system.

Recording Human Electrocorticographic ECoG Signals for Neuroscientific Research and Real-time Functional Cortical Mapping

We present a method for collecting electrocorticographic signals for research purposes from humans who are undergoing invasive epilepsy monitoring. We show how to use the BCI2000 software platform for data collection, signal processing and stimulus presentation. Specifically, we demonstrate SIGFRIED, a BCI2000-based tool for real-time functional brain mapping.

神经科学的研究和实时功能皮层映射记录人力Electrocorticographic（ECOG）信号

Neuroimaging studies of human cognitive, sensory, and motor processes are usually based on noninvasive techniques such as electroencephalography (EEG), ...

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 ...

Microdialysis of Ethanol During Operant Ethanol Self-administration and Ethanol Determination by Gas Chromatography

Operant self-administration methods are commonly used to study the behavioral and pharmacological effects of many drugs of abuse, including ethanol. However, ethanol is typically self-administered orally, rather than intravenously like many other drugs of abuse. The pharmacokinetics of orally administered drugs are more complex than intravenously administered drugs. Because understanding the relationship between the pharmacological and behavioral effects of ethanol requires knowledge of the time course of ethanol reaching the brain during and after drinking, we use in vivo microdialysis and gas chromatography with flame ionization detection to monitor brain dialysate ethanol concentrations over time. 
Combined microdialysis-behavioral experiments involve the use of several techniques. In this article, stereotaxic surgery, behavioral training and microdialysis, which can be adapted to test a multitude of self-administration and neurochemical centered hypotheses, are included only to illustrate how they relate to the subsequent phases of sample collection and dialysate ethanol analysis. Dialysate ethanol concentration analysis via gas chromatography with flame-ionization detection, which is specific to ethanol studies, is described in detail. Data produced by these methods reveal the pattern of ethanol reaching the brain during the self-administration procedure, and when paired with neurochemical analysis of the same dialysate samples, allows conclusions to be made regarding the pharmacological and behavioral effects of ethanol.

A method to determine the time course of ethanol concentration in the brains of rats during operant ethanol self-administration is described. Gas chromatography with flame ionization detection is used to quantify ethanol in the dialysate samples, because it has the sensitivity required for the small volumes that are generated.

微透析的乙醇操作性乙醇的自我管理和乙醇的测定气相色谱

Operant self-administration methods are commonly used to study the behavioral and pharmacological effects of many drugs of abuse, including ethanol.  ...

Simultaneous Electroencephalography, Real-time Measurement of Lactate Concentration and Optogenetic Manipulation of Neuronal Activity in the Rodent Cerebral Cortex

Although the brain represents less than 5% of the body by mass, it utilizes approximately one quarter of the glucose used by the body at rest1. The function of non rapid eye movement sleep (NREMS), the largest portion of sleep by time, is uncertain. However, one salient feature of NREMS is a significant reduction in the rate of cerebral glucose utilization relative to wakefulness2-4. This and other findings have led to the widely held belief that sleep serves a function related to cerebral metabolism. Yet, the mechanisms underlying the reduction in cerebral glucose metabolism during NREMS remain to be elucidated.
	One phenomenon associated with NREMS that might impact cerebral metabolic rate is the occurrence of slow waves, oscillations at frequencies less than 4 Hz, in the electroencephalogram5,6. These slow waves detected at the level of the skull or cerebral cortical surface reflect the oscillations of underlying neurons between a depolarized/up state and a hyperpolarized/down state7. During the down state, cells do not undergo action potentials for intervals of up to several hundred milliseconds. Restoration of ionic concentration gradients subsequent to action potentials represents a significant metabolic load on the cell8; absence of action potentials during down states associated with NREMS may contribute to reduced metabolism relative to wake.
	Two technical challenges had to be addressed in order for this hypothetical relationship to be tested. First, it was necessary to measure cerebral glycolytic metabolism with a temporal resolution reflective of the dynamics of the cerebral EEG (that is, over seconds rather than minutes). To do so, we measured the concentration of lactate, the product of aerobic glycolysis, and therefore a readout of the rate of glucose metabolism in the brains of mice. Lactate was measured using a lactate oxidase based real time sensor embedded in the frontal cortex. The sensing mechanism consists of a platinum-iridium electrode surrounded by a layer of lactate oxidase molecules. Metabolism of lactate by lactate oxidase produces hydrogen peroxide, which produces a current in the platinum-iridium electrode. So a ramping up of cerebral glycolysis provides an increase in the concentration of substrate for lactate oxidase, which then is reflected in increased current at the sensing electrode. It was additionally necessary to measure these variables while manipulating the excitability of the cerebral cortex, in order to isolate this variable from other facets of NREMS.
	We devised an experimental system for simultaneous measurement of neuronal activity via the elecetroencephalogram, measurement of glycolytic flux via a lactate biosensor, and manipulation of cerebral cortical neuronal activity via optogenetic activation of pyramidal neurons. We have utilized this system to document the relationship between sleep-related electroencephalographic waveforms and the moment-to-moment dynamics of lactate concentration in the cerebral cortex. The protocol may be useful for any individual interested in studying, in freely behaving rodents, the relationship between neuronal activity measured at the electroencephalographic level and cellular energetics within the brain.

A procedure is described for manipulating the activity of cerebral cortical pyramidal neurons optogenetically while the electroencephalogram, electromyogram, and cerebral lactate concentration are monitored. Experimental recordings are performed on cable-tethered mice while they undergo spontaneous sleep/wake cycles. Optogenetic equipment is assembled in our laboratory; recording equipment is commercially available.

同时脑电图，实时测量的啮齿动物的大脑皮质中的神经元活动的乳酸浓度与Optogenetic的操作

Although the brain represents less than 5% of the body  by mass, it utilizes approximately one quarter of the glucose used by the body  at rest1. The ...

A Simple One-step Dissection Protocol for Whole-mount Preparation of Adult Drosophila Brains

There is an increasing interest in using Drosophila to model human brain degenerative diseases, map neuronal circuitries in adult brains, and study the molecular and cellular basis of higher brain functions. A whole-mount preparation of adult brains with well-preserved morphology is critical for such whole brain-based studies, but can be technically challenging and time-consuming. This protocol describes an easy-to-learn, one-step dissection approach of an adult fly head in less than 10 s, while keeping the intact brain attached to the rest of the body to facilitate subsequent processing steps. The procedure helps remove most of the eye and tracheal tissues normally associated with the brain that can interfere with the later imaging step, and also places less demand on the quality of the dissecting forceps. Additionally, we describe a simple method that allows convenient flipping of the mounted brain samples on a coverslip, which is important for imaging both sides of the brains with similar signal intensity and quality. As an example of the protocol, we present an analysis of dopaminergic (DA) neurons in adult brains of WT (w1118) flies. The high efficacy of the dissection method makes it particularly useful for large-scale adult brain-based studies in Drosophila.

A Simple One-step Dissection Protocol for Whole-mount Preparation of Adult Drosophila Brains

The adult Drosophila brain is a valuable system for studying neuronal circuitry, higher brain functions, and complex disorders. An efficient method to dissect whole brain tissue from the small fly head will facilitate brain-based studies. Here we describe a simple, one-step dissection protocol of adult brains with well-preserved morphology.

一个简单的一步解剖协议成人全安装准备<em&gt;果蝇</em&gt;脑

There is an increasing interest in using Drosophila to model human brain degenerative diseases, map neuronal circuitries in adult brains, and study the ...

Assay to Measure Nucleocytoplasmic Transport in Real Time within Motor Neuron-like NSC-34 Cells

Nucleocytoplasmic transport refers to the import and export of large molecules from the cell nucleus. Recently, a number of studies have shown a connection between amyotrophic lateral sclerosis (ALS) and impairments in the nucleocytoplasmic pathway. ALS is a neurodegenerative disease affecting the motor neurons and resulting in paralysis and ultimately in death, within 2-5 years on average. Most cases of ALS are sporadic, lacking any apparent genetic linkage, but 10% are inherited in a dominant manner. Recently, hexanucleotide repeat expansions (HREs) in the chromosome 9 open reading frame 72 (C9orf72) gene were identified as a genetic cause of ALS and frontotemporal dementia (FTD). Importantly, different groups have recently proposed that these mutants affect nucleocytoplasmic transport. These studies have mostly shown the final outcome and manifestations caused by HREs on nucleocytoplasmic transport, but they do not demonstrate nuclear transport dysfunction in real time. As a result, only severe nucleocytoplasmic transport deficiency can be determined, mostly due to high overexpression or exogenous protein insertion. 
This protocol describes a new and very sensitive assay to evaluate and quantify nucleocytoplasmic transport dysfunction in real time. The rate of import of a NLS-NES-GFP protein (shuttle-GFP) can be quantified in real time using fluorescent microscopy. This is performed by using an exportin inhibitor, thus allowing the shuttle GFP only to enter the nucleus. To validate the assay, the C9orf72 HRE translated dipeptide repeats, poly(GR) and poly(PR), which have been previously shown to disrupt nucleocytoplasmic transport, were used. Using the described assay, a 50% decrease in the nuclear import rate was observed compared to the control. Using this system, minute changes in nucleocytoplasmic transport can be examined and the ability of different factors to rescue (even partially) a nucleocytoplasmic transport defect can be determined.

This protocol describes a method to sensitively measure the nucleocytoplasmic transport rate within motor neuron-like NSC-34 cells by quantifying the real-time change in the nuclear import of a NLS-NES-GFP protein.

实时 fMRI 神经训练管理协议

摘要

探索更多视频

此视频中的章节