Name: assessing pupil-linked changes in locus coeruleus-mediated arousal elicited by trigeminal stimulation
Uploaded: 2019-11-26T12:30:00
Description: Watch this Scientific Journal Video about Assessing Pupil-linked Changes in Locus Coeruleus-mediated Arousal Elicited by Trigeminal Stimulation at JoVE.com

这项研究得到了比萨大学的资助。我们感谢保罗·奥尔西尼先生、弗朗切斯科·蒙塔纳里先生和克里斯蒂娜·普奇夫人的宝贵技术援助，并感谢国际投资公司S.R.R.L.公司为玛丽亚·保拉·特拉蒙蒂·范托齐博士提供奖学金。最后，我们感谢 OCM 项目公司准备硬颗粒并执行硬度和弹簧恒定测量。

所有步骤都遵循比萨大学道德委员会的指导方针。
1. 参与者招聘
<ol><li>根据研究的具体目标（即正常受试者和/或患者、男性和/或女性、年轻人和/或老年人）招募受试者群体。</li>
</ol>2. 材料准备
<ol><li>准备软颗粒;使用市售口香糖（材料表;初始硬度=20肖氏OO）。</li>
	<li>准备硬颗粒;使用硅橡胶颗粒（材料表;恒定硬度= 60肖氏OO）<s...

<ol>
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本研究提出的方案涉及感觉运动三叉体活动对认知性能的急性影响以及LC在这一过程中的作用。这个话题有一定的相关性，考虑到1）在老化期间，咀嚼活性的恶化与认知衰减32，33，34;维护口腔健康的人不太容易出现神经退行性现象;2） 闭塞和牙齿提取诱导神经退行性作用在动物在海马和皮质水平35，<...

在人类中，众所周知，咀嚼会加速认知处理1，2，并改善觉醒3，4，注意5，学习和记忆6，7。这些影响与缩短皮质事件相关电位8的延迟和增加几个皮质和皮下结构2，9的灌注增加有关。
在颅神经内，最相关的信息维持皮质去同步和唤醒是由三叉纤维10携带，可能是由于强烈的三叉线连接到上升的视网膜激活系统（ARAS）11 。在ARAS结构中，球位（LC）接收三元位输入11，调制唤醒12，13，其活动与学生大小14，15，16，17，18。虽然LC休息活动与认知表现之间的关系是复杂的，但与任务相关的LC活动的增强导致觉醒相关的19名学生心肌炎20和增强认知性能21。LC活动与学生大小之间存在可靠的共性差异，后者目前被认为是22、23、24、25、26的中央诺瑞纳活性的代理。
传感器运动三叉叶分支的不对称激活诱导学生不对称（双子座）27，28，确认三角-科鲁里亚连接的强度。如果LC参与咀嚼对认知表现的刺激作用，它可能会影响平行的任务相关的突起症，这是在任务期间LC相发生数激活的指标。它还可能影响性能，因此，咀嚼引起的性能变化与支数症之间存在相关性。此外，如果三叉体效应是特定的，咀嚼效果应该大于那些由另一个有节奏的运动任务引起的。为了检验这些假设，特此提出了两个实验方案。它们基于认知表现和学生大小的综合测量，在短暂的咀嚼活动前后进行。这些协议利用一个测试，包括查找数字关注矩阵29中显示的目标数，以及非目标数字。此测试验证专注和认知性能。
这些协议的总体目标是说明三元刺激会引起认知性能的具体变化，这不能具体归因于电机命令的生成，并且与LC介导的与学生相关的变化有关觉醒。该协议的应用扩展到所有行为条件，其中可以测量性能，并怀疑 LC 的参与。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Anti-stress ball</td>
			<td>Artengo, Decathlon, France</td>
			<td>TB600</td>
			<td></td>
		</tr>
		<tr>
			<td>Chewing gum</td>
			<td>Vigorsol, Perfetti, Italy</td>
			<td>Commercially available product</td>
			<td></td>
		</tr>
		<tr>
			<td>Infrared Camera-Wearable pupillometer</td>
			<td>Pupil Labs, Berlin, Germany</td>
			<td>Pupil Labs headset</td>
			<td></td>
		</tr>
		<tr>
			<td>Pupillographer</td>
			<td>CSO, Florence, Italy</td>
			<td>MOD i02, with chin support</td>
			<td></td>
		</tr>
		<tr>
			<td>Silicon rubber</td>
			<td>Prochima, Italy</td>
			<td>gls50</td>
			<td></td>
		</tr>
		<tr>
			<td>Software for pupil detection - wearable pupillometer</td>
			<td>Pupil Labs, Berlin, Germany</td>
			<td>Pupil Labs headset</td>
			<td></td>
		</tr>
		<tr>
			<td>Tangram Puzzle</td>
			<td>Citt&#224; del Sole srl, Milano, Italy</td>
			<td>Tangram Puzzle</td>
			<td></td>
		</tr>
		<tr>
			<td>Wearable pupillometer</td>
			<td>Pupil Labs, Berlin, Germany</td>
			<td>Pupil labs model</td>
			<td>Dimension of the frame: 13.5 x 15.5cm</td>
		</tr>
	</tbody>
</table>

assessing pupil-linked changes in locus coeruleus-mediated arousal elicited by trigeminal stimulation

目前的科学文献提供了证据，证明与咀嚼相关的三叉体感觉运动活动可能会影响觉醒、注意力和认知表现。这些影响可能是由于三叉星系统与上升性视网膜激活系统 （ARAS） 的广泛连接，而基拉图（LC）的角质神经元属于这个系统。LC神经元包含对整个大脑的投影，并且已知它们的放电随学生大小共同变化。LC 激活对于引发与任务相关的突起体炎是必要的。如果咀嚼对认知性能的影响由LC调节，那么有理由期望认知性能的变化与任务相关突起系统的变化相关。这里提出了两种新颖的协议来验证这一假设，并证明咀嚼效果不能归因于特定的运动激活。在这两种协议中，在特定任务期间观察到的性能和学生大小变化都会在 2 分钟之后记录在 2 分钟之后和半小时内记录：a） 无活动、b） 有节奏、双边握把、c） 双边咀嚼软颗粒和 d）双边咀嚼硬颗粒。第一个协议测量在数字矩阵中显示的目标数字的识别性能级别。由于学生大小记录由适当的学生表记录，该测速仪会妨碍视力以确保恒定的照明水平，因此在触觉任务期间会评估与任务相关的霉菌病。该协议的结果表明，1） 咀嚼引起的性能和任务相关的突起病的变化是相关的，2） 性能和突动都不能通过手柄增强。在第二个协议中，使用可穿戴的孔径计可以测量同一任务期间的学生大小变化和性能，从而获得关于LC参与认知活动三元效应的更有力的证据。这两种协议都运行在比萨大学ARAS的发现者朱塞佩·莫鲁齐教授的历史办公室。

图 4显示了将协议 1 应用于单个主体（46 岁，女性）时获得的结果的代表性示例。PI 在咀嚼 （T7） 后很快增加（从 1.73 麻木/s 到 2.27 麻木/s） 和软颗粒 （从 1.67 麻木/s 到 1.87 麻木/s） （图 4A）。然而，30分钟后（T37），增加的性能只坚持硬颗粒。另一方面，缺乏活动和握把运动对性能有负面影响，从1.73麻木/秒下降到1.67麻木/秒，从1.6麻木/...

Watch this Scientific Journal Video about Assessing Pupil-linked Changes in Locus Coeruleus-mediated Arousal Elicited by Trigeminal Stimulation at JoVE.com

Assessing Pupil-linked Changes in Locus Coeruleus-mediated Arousal Elicited by Trigeminal Stimulation

为了验证三元对认知表现的影响是否涉及位点性球菌活动，提出了两个协议，旨在评估咀嚼引起的性能和与任务相关的学生大小变化之间的可能相关性。这些协议可应用于怀疑原点贡献的条件。

评估由三角刺激引起的由Locus Coeruleus介导的觉醒中与学生相关的变化

Current scientific literature provides evidence that trigeminal sensorimotor activity associated with chewing may affect arousal, attention, and cognitive ...

蝗虫是一种深层大脑结构，调节行为觉醒、注意力和认知表现。通过测量瞳孔大小，可以间接记录其活动。这种方法允许验证促进认知性能的刺激（如 Gemina 刺激）的用，随着瞳孔尺寸的直径变化调节 LC 活性。实验证据表明，神经退行性疾病与位子库瘤功能障碍有关。目前的方法将为将宝石输入调制到蝗虫库进行治疗目的铺平道路。与托马索·班菲一起演示这个程序的将是文森佐·德·西科，一位专业医生和眼科医生。为了进行实验，需要一种商业上可用的口香糖，称为软颗粒，以及一种硅橡胶颗粒，称为硬颗粒。还必须为触觉任务准备一个唐拉姆拼图。对于协议一，提供一篇论文，向主体显示三个 10 到 10 个数值矩阵，并要求主体在 15 秒内按顺序扫描矩阵行，同时使用铅笔勾选每个矩阵上方指示的尽可能多的目标数字，以测量主体的基线认知性能。然后手动评估性能指标、扫描速率和错误率。要测量基线瞳孔大小，请将主体从角膜测地仪的 56 毫米的最佳工作距离放在 56 毫米左右的瞳孔上，并在 40 lux 的恒定照明下获取左右瞳孔的单独相机拍摄。要评估在 Tangram 触觉任务期间的瞳孔大小，请将拼图中一块放入主体的手中，并要求主体在记录主体瞳孔尺寸的同时将棋子返回给橘子。要测量触觉任务期间的瞳孔大小，请获取照片，而主体在拼图表面探索开始时执行两个任务重复中的第二个。通过直接获取软件显示的值（以毫米为单位）来评估左右瞳孔尺寸。然后在触觉任务期间从瞳孔大小中减去瞳孔大小，以计算与任务相关的霉菌病，并获取右侧和左侧瞳孔的平均值。接下来，要求主体咀嚼一个自我管理的软颗粒，让主体自发地选择咀嚼速率和首选的咀嚼侧。一分钟后，为主体提供新的软颗粒，并要求主体切换咀嚼侧再咀嚼一分钟。紧接着，结束咀嚼练习，评估主体在矩阵测试中的表现，在测量瞳孔大小时，在休息和触觉任务期间。咀嚼练习结束后 30 分钟，在休息和触觉任务期间评估主体性能和瞳孔大小。对于协议二，为主体配备具有 3D 打印玻璃框架结构的可穿戴瞳孔眼动镜，并调整安装在框架上的两个红外摄像机的位置，使主体的眼睛与摄像机的视野对焦。使用安装在可穿戴瞳孔框架上的校准对数光传感器连续同时记录环境照明水平，在可穿戴瞳孔软件内以 120 赫兹采样率获取受器在其余位置的图像，持续 20 秒。要获取基线认知性能数据，请记录学生的大小，而主体执行 Spinnler-Tognoni 测试。手动提升左右瞳孔尺寸，在 Spinnler-Tognoni 测试期间，通过平均每个瞳孔的获取值。然后计算与任务相关的菌体，如所证明的，以获得左右瞳孔的平均值。接下来，要求主体咀嚼一个自我管理的软颗粒，让主体自发地选择咀嚼速率和咀嚼嘴侧。一分钟后，为主体提供新的软颗粒，并要求主体切换咀嚼侧，再咀嚼一分钟。在咀嚼练习结束时，立即评估在矩阵测试期间在性能和瞳孔尺寸方面处于休息的瞳孔大小。咀嚼练习结束后 30 分钟，在矩阵测试期间评估瞳孔大小和性能瞳孔大小。在这个具有代表性的示例中，在咀嚼了硬颗粒或软颗粒后，性能指数很快提高。然而，30 分钟后，性能的提高只持续对于硬颗粒。两种控制条件，如缺乏活动和手握运动，对性能有负面影响，30分钟后有恢复的趋势。在上一个图中显示的同一主题中，对任务相关的菌体疾病观察到了质量上类似的变化。在仪器的连续采集模式下，最终的单个样品代表最终平均值，因为瞳孔在关灯后五秒内达到稳定尺寸。在单个受试者中，在咀嚼硬颗粒和软颗粒后，观察到性能与任务相关的性病菌之间有很强的相关性。当对基线值进行相应的更改规范化时，相关性也很明显。通过将咀嚼引起的性能指数变化与执行同一矩阵测试期间观察到的肌病的变化关联，可以获得更强有力的证据，证明 LC 参与咀嚼对认知性能的刺激作用。使用该程序，仔细控制环境照明非常重要，因为外部光线可能是瞳孔尺寸的主要决定因素，混淆了结果。此过程可能会发现几个应用程序，例如，我们目前正在研究对闭塞亚米法的校正是否会通过影响位子库活动来提高性能。

Research

JoVE Journal

Neuroscience

TMS: Using the Theta-Burst Protocol to Explore Mechanism of Plasticity in Individuals with Fragile X Syndrome and Autism

TMS：使用的Theta突发协议，以探索个人与脆性X综合征与孤独症的可塑性Mechasnism

Fragile X Syndrome (FXS), also known as Martin-Bell Syndrome, is a genetic abnormality found on the X chromosome.1,2 Individuals suffering from FXS ...

科研

神经科学

Assessing Neurodegenerative Phenotypes in Drosophila Dopaminergic Neurons by Climbing Assays and Whole Brain Immunostaining

Assessing Neurodegenerative Phenotypes in Drosophila Dopaminergic Neurons by Climbing Assays and Whole Brain Immunostaining

评估神经退行性表型<em&gt;果蝇</em&gt;爬坡试验和全脑多巴胺能神经元免疫组化染色

Drosophila melanogaster is a valuable model organism to study aging and pathological degenerative processes in the nervous system. The advantages of the ...

Deep Brain Stimulation with Simultaneous fMRI in Rodents

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 ...

Analysis of Gene Expression Changes in the Rat Hippocampus After Deep Brain Stimulation of the Anterior Thalamic Nucleus

基因表达的变化大鼠海马后，前丘脑核脑深部刺激分析

Deep brain stimulation (DBS) surgery, targeting various regions of the brain such as the basal ganglia, thalamus, and subthalamic regions, is an effective ...

Generation of Local CA1 &#947; Oscillations by Tetanic Stimulation

Generation of Local CA1 ÃƒÅ½Ã‚Â³ Oscillations by Tetanic Stimulation

本地CA1γ振荡通过强直刺激生成

Neuronal network oscillations are important features of brain activity in health and disease and can be modulated by a range of clinically used drugs. A ...

Effects of Transcranial Alternating Current Stimulation on the Primary Motor Cortex by Online Combined Approach with Transcranial Magnetic Stimulation

经颅磁刺激在线联合方法对经颅交流电刺激对原发性运动皮层的影响

Transcranial Alternating Current Stimulation (tACS) is a neuromodulatory technique able to act through sinusoidal electrical waveforms in a specific ...

Ocular Kinematics Measured by In Vitro Stimulation of the Cranial Nerves in the Turtle

Ocular Kinematics Measured by In Vitro Stimulation of the Cranial Nerves in the Turtle

体外刺激对龟脑神经的运动测量

After animals are euthanized, their tissues begin to die. Turtles offer an advantage because of a longer survival time of their tissues, especially when ...

Localization of the Locus Coeruleus in the Mouse Brain

The locus coeruleus (LC) is a major hub of norepinephrine producing neurons that modulate a number of physiological functions. Structural or functional ...

Assessing Early Stage Open-Angle Glaucoma in Patients by Isolated-Check Visual Evoked Potential

通过隔离检查视觉诱发潜力评估患者早期开放角青光眼

Recently, the isolated-check visual evoked potential (icVEP) technique was designed and has been reported to detect glaucomatous damage earlier and ...

Establishment of Acute Pontine Infarction in Rats by Electrical Stimulation

电刺激在大鼠中建立急性庞廷梗死

Pontine infarction is the most common stroke subtype in the posterior circulation, while there lacks a rodent model mimicking pontine infarction. Provided ...