这项工作得到了京都大学白尾项目、挑战研究补助金（先锋）（No.22K18644）、科学研究补助金 （C） （No. 22K12745）、科学研究补助金 （B） （No. 21H02851） 和科学研究补助金 （A） （No. 19H01039） 的支持。感谢 意得辑 （www.editage.jp） 的英文编辑工作。

所有实验均经 EHUB 动物实验委员会（No.2022-003、2023-104）批准，并按照 EHUB 发布的《实验室灵长类动物护理和使用指南》进行。实验使用了 9 只普通狨猴 （Callithrix jacchus，6 只雄性和 3 只雌性，2-12 岁，体重 330-490 g）。
1. 动物
<ol><li>将狨猴安置在配备巢箱、木制栖木和其他丰富装置的单个笼子中。</li>
	<li>将房间保持在 12 小时的光暗循环下，温度和湿度分...

用于狨猴声音感知比较研究的无创脑电图

作者没有需要披露的利益冲突。

关于麻醉的注意事项 氯胺酮和甲苯噻嗪给药都已被尝试过，虽然它们是镇痛剂，因此适用于长时间的痛苦任务，但狨猴在没有吸氧的情况下往往会出现血氧水平降低44。简而言之，alfaxalon 可能最适合剃须或制作面膜等无痛任务。此外，对于仅需 10-15 分钟的剃须，吸入麻醉将是最合适的。由于异氟醚持续时间短且浓度低 （约 1%），因此在插管期间未使用。
<p cla...

灵长类动物使用物种特异性的发声来传达生物学上重要的信息，例如呼叫者的情绪状态或维持社会纽带的意图、捕食者的存在或其他危险情况。对声音丰富的非人类灵长类动物发声感知背后的神经机制的研究可能为我们提供关键线索，以更好地理解人类语言的进化起源。
普通狨猴是原产于南美洲的小型灵长类动物。近年来，狨猴越来越多地与猕猴一起被用作模式动物，因为它们的繁殖能力高，体型小，易于使用，以及有用的转基因技术的发展 1,2,3。除了作为疾病模型的实用性外，群体内丰富的声音交流是该物种的另一个独特特征 4,5,6,7。狨猴经常交换声音信号，与森林中看不见的同种动物交流。通过检查狨猴发声感知和产生所涉及的大脑活动，我们可以确定它们如何处理大脑中自身或同种呼叫的听觉信息，并确定涉及哪些神经回路。先前的研究表明，初级听觉皮层8、9、10、11、12 和额叶皮层13、14 的神经活动参与狨猴的声音产生。此外，这些兴奋和抑制的神经元反应受到初级听觉皮层中听觉-声音相互作用的调节 8,10。这些研究使用侵入性记录方法提供了单神经元水平的详细神经活动数据。许多研究进一步检查了狨猴发声产生所涉及的神经活动;然而，对声音感知的理解仍然知之甚少15,16。
几项无创脑成像研究阐明了狨猴发声处理的神经机制 17,18,19;它们的高空间分辨率是一个优势，但是，在扫描过程中使动物保持清醒状态需要先进的技术。然而，最近，Jafari 等人使用功能性磁共振成像 （fMRI） 确定了清醒狨猴参与声音感知的额颞区19。几乎所有阐明人类参与发声感知和产生的大脑功能的实验都是使用无创方法进行的，例如头皮脑电图 （EEG）、脑磁图 （MEG）20,21 和 fMRI 22,23,24。许多人类研究调查了使用 EEG 与声音感知相关的大脑活动。这些研究大多集中在情绪信息 25,26,27 和情绪词的显著性 28 上，结果揭示了声音感知过程中事件相关电位的变化29。使用颅内植入电极的人体皮层电图 （ECoG） 和单神经元记录仅在接受神经外科治疗的患者中进行了有限数量的实验30,31。
在了解人类发声感知和产生背后的独特神经机制时，将人类与猴子进行比较的进化观点非常重要。为了直接比较声音丰富的非人类灵长类动物（如狨猴）与人类的语音感知和发声所涉及的神经机制，使用相同的方法比较两个物种之间的数据非常重要。功能性 MRI 允许全脑成像并具有很高的空间分辨率。它的优点是可以记录垂直于颅骨或难以用 EEG 或 MEG 记录的深部区域的活动。然而，MRI 机器的安装和维护成本很高，并且由于设备的性质，可以呈现的刺激存在许多限制。相比之下，EEG、事件相关电位 （ERP） 和 MEG 具有较高的时间分辨率，使其可用于分析时间序列声音处理。特别是，脑电图具有移动性高、能够在各种实验环境中使用、成本相对较低以及只需要一名操作员的优势。
由于已经在人类身上获得了大量的脑电图数据，因此非人类灵长类动物需要使用非侵入性范式的脑电图测量方法。我们的研究小组开发了一种独特的无创脑电图记录方法，使用32 号管对猕猴和狨猴进行记录。在这里，我们报告了有关非人灵长类动物听觉处理的几项新发现 33,34,35,36,37。为了表征响应狨猴物种特异性发声的大脑活动，我们构建了一个实验系统，使用放置在头皮上的电极无创记录大脑活动。在这项研究中，我们描述了狨猴的脑电图测量方法。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Alfaxalone</td>
			<td>Meiji Animal Health</td>
			<td>Alfaxan</td>
			<td></td>
		</tr>
		<tr>
			<td>Amplifier</td>
			<td>Brain Products</td>
			<td>BrainAmp</td>
			<td></td>
		</tr>
		<tr>
			<td>Atropine</td>
			<td>Fuso Pharmaceutical Industries</td>
			<td>Atropine Sulfate Injection</td>
			<td></td>
		</tr>
		<tr>
			<td>Audio editor</td>
			<td>Adobe</td>
			<td>Adobe Audition</td>
			<td></td>
		</tr>
		<tr>
			<td>Data processing software</td>
			<td>MathWorks</td>
			<td>MATLAB</td>
			<td>version R2023a</td>
		</tr>
		<tr>
			<td>Data processing toolbox</td>
			<td>University of California-SanDiego</td>
			<td>EEGLAB</td>
			<td></td>
		</tr>
		<tr>
			<td>Data processing toolbox</td>
			<td>University of California-Davis</td>
			<td>ERPLAB</td>
			<td></td>
		</tr>
		<tr>
			<td>Electric shaver</td>
			<td>Panasonic</td>
			<td>ER803PPA</td>
			<td></td>
		</tr>
		<tr>
			<td>Electrode</td>
			<td>Unique Medical</td>
			<td>UL-3010</td>
			<td>AgCl coated (custom)</td>
		</tr>
		<tr>
			<td>Electrode gel</td>
			<td>Neurospec AG</td>
			<td>V16 SuperVisc</td>
			<td></td>
		</tr>
		<tr>
			<td>Electrode input box</td>
			<td>Brain Products</td>
			<td>EIB64-DUO</td>
			<td>64ch</td>
		</tr>
		<tr>
			<td>Glue</td>
			<td>3M</td>
			<td>Scotch 7005S</td>
			<td></td>
		</tr>
		<tr>
			<td>Hair removering cream</td>
			<td>Kracie</td>
			<td>epilat for sensitive skin</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Bussan Animal Health</td>
			<td>ds isoflurane</td>
			<td></td>
		</tr>
		<tr>
			<td>Liquid gum</td>
			<td>San-ei Yakuhin Boeki</td>
			<td>Arabic Call SS</td>
			<td>Gum arabic+water</td>
		</tr>
		<tr>
			<td>Liquid nutrition</td>
			<td>Nestl&#233; Health Science Company</td>
			<td>Isocal 1.0 Junior</td>
			<td>Polymeric formula</td>
		</tr>
		<tr>
			<td>Maropitant</td>
			<td>Zoetis&#160;</td>
			<td>Cerenia injectable solution</td>
			<td></td>
		</tr>
		<tr>
			<td>Monitor Camera</td>
			<td>Intel</td>
			<td>RealSense LiDAR Camera L515</td>
			<td></td>
		</tr>
		<tr>
			<td>Monkey pellets</td>
			<td>Oriental Yeast</td>
			<td>SPS</td>
			<td></td>
		</tr>
		<tr>
			<td>Primate chair</td>
			<td>Natsume Seisakusho</td>
			<td>Order made</td>
			<td></td>
		</tr>
		<tr>
			<td>Pulse oximeters</td>
			<td>Covident</td>
			<td>Nellcor</td>
			<td>PM10N</td>
		</tr>
		<tr>
			<td>Skin prepping pasta&#160;</td>
			<td>Mammendorfer Institut f&#252;r Physik und Medizin</td>
			<td>NeuPrep</td>
			<td></td>
		</tr>
		<tr>
			<td>Slicon tube</td>
			<td>AsONE</td>
			<td>&#934;4 x 7mm</td>
			<td></td>
		</tr>
		<tr>
			<td>Speaker</td>
			<td>Fostex</td>
			<td>PM0.3</td>
			<td></td>
		</tr>
		<tr>
			<td>Synchronization device</td>
			<td>Brain Vision</td>
			<td>StimTrak</td>
			<td></td>
		</tr>
		<tr>
			<td>Thermoplastic mask</td>
			<td>CIVCO</td>
			<td>MTAPU Type Uniframe Thermoplastic Mask 2.4mm</td>
			<td></td>
		</tr>
	</tbody>
</table>

electroencephalography measurements in awake marmosets listening to conspecific vocalizations 

声音交流在灵长类动物的社交互动中起着至关重要的作用，尤其是在生存和社会组织中。人类以语言的形式发展出一种独特而先进的声音交流策略。要研究人类语言的进化，有必要研究人类发声处理背后的神经机制，并通过将它们与非人类灵长类动物的机制进行比较来了解大脑机制是如何进化的。在此，我们开发了一种无创测量清醒非人灵长类动物脑电图 （EEG） 的方法。这种记录方法允许在不伤害动物的情况下进行长期研究，重要的是，它允许我们直接将非人类灵长类动物的脑电图数据与人类数据进行比较，从而深入了解人类语言的进化。在目前的研究中，我们使用头皮脑电图记录方法来研究大脑活动对狨猴物种特异性发声的反应。本研究通过使用头皮脑电图捕捉狨猴在声音感知过程中的广泛神经表征，填补了现有知识的空白，从而提供了新的见解。

Primates use species-specific vocalizations to convey biologically important information, such as the caller&#39;s emotional state or intention to maintain social bonds, the presence of predators, or other dangerous situations. Investigation of the neural mechanisms underlying the perception of vocalization in vocal-rich nonhuman primates may provide us with critical clues to better understand the evolutionary origins of human language.
Common marmosets are small primates native to South America. In recent years, marmosets have been increasingly used as model animals, alongside macaque monkeys, because of their high reproductivity, ease of use owing to their small size, and the development of useful transgenic techniques1,2,3. In addition to their utility as disease models, rich vocal communication within groups is another unique characteristic of this species4,5,6,7. Marmosets routinely exchange vocal signals to communicate with invisible conspecifics in the forest. By examining the brain activity involved in vocal perception and production in marmosets, we can determine how they process the auditory information of their own or conspecific calls in the brain and identify which neural circuits are involved. Previous studies have demonstrated neural activity in the primary auditory cortex8,9,10,11,12 and frontal cortex13,14 involved in vocal production in marmosets. Furthermore, these excited and suppressed neuronal responses were modulated by auditory-vocal interactions in the primary auditory cortex8,10. These studies provided detailed neural activity data at the single-neuron level using invasive recording methods. Numerous studies have further examined the neural activity involved in marmoset vocal production; however, vocal perception remains poorly understood15,16.
Several noninvasive brain imaging studies have elucidated the neural mechanisms of vocal processing in marmosets17,18,19; their high spatial resolution is an advantage, however, keeping animals in the awake state during scanning requires advanced techniques. However, more recently, Jafari et al. identified frontotemporal regions involved in vocal perception in awake marmosets using functional magnetic resonance imaging (fMRI)19. Almost all experiments to elucidate the brain functions involved in vocal perception and production in humans have been conducted using noninvasive methods, such as scalp electroencephalography (EEG), magnetoencephalography (MEG)20,21, and fMRI22,23,24. Numerous studies in humans have investigated brain activity related to vocal perception using EEG. Most of these studies have focused on emotional information25,26,27 and the saliency of emotional words28, with the results revealing changes in event-related potentials during vocal perception29. Electrocorticography (ECoG) and single-neuron recordings using intracranially implanted electrodes in humans have only been conducted in a limited number of experiments in patients undergoing neurosurgical treatment30,31.
An evolutionary perspective comparing humans with monkeys is important when understanding the unique neural mechanisms underlying vocal perception and production that have developed in humans. To directly compare the neural mechanisms involved in speech perception and vocalization in vocal-rich nonhuman primates, such as the marmoset, with humans, it is important to compare data between the two species using the same method. Functional MRI allows whole-brain imaging and has a high spatial resolution. It has the advantage of recording activity perpendicular to the skull or in deep regions that are difficult to record with EEG or MEG. However, the MRI machine is expensive to install and maintain, and there are many restrictions on the stimuli that can be presented due to the nature of the device. In comparison, EEG, event-related potentials (ERPs), and MEG have a high temporal resolution, making them useful for analyzing time-series vocal processing. In particular, EEG has the advantages of high mobility and the ability to be used in a variety of experimental settings, relatively low cost, and the requirement for just a single operator.
Since a large amount of EEG data has already been obtained in humans, EEG measurement methods using non-invasive paradigms are needed for non-human primates. Our research group developed a unique noninvasive EEG recording method using tubes32 for macaques and marmosets. Here, we report several novel findings regarding auditory processing in nonhuman primates33,34,35,36,37. To characterize brain activity in response to species-specific vocalizations in marmosets, we constructed an experimental system to noninvasively record brain activity using electrodes placed on the scalp. In this study, we describe the EEG measurement method for marmosets.

作者聚焦：使用无创脑电图研究小型灵长类动物的声音信息表示及其精神疾病对声音信息的改变

清醒狨猴的脑电图测量：聆听同种发声 

We would like to know how vocal information is represented in the brain of small primates which are rich in vocal communications. We would like to illustrate how the representation will be altered by psychiatric disorders such as schizophrenia in the future. We compared non-invasive EEGs across four primate species, and found that the latency of auditory cortex to respond to sounds is longer in humans than in other primates.This would have a significant impact on the perception of complex sounds, such as calls and speech sounds. Few experimental methods allow direct comparison of brain activity obtained from human and animals. We hope to quantify the process of human language evolution by direct comparison, or directly comparing data among various animal species using the non-invasive measurement method.We hope to address this challenge with our protocol.

首先，我们绘制了狨猴中每种听觉刺激的平均事件相关电位 （ERP）（图 2）。听觉诱发电位 （AEP） 在 噪声 条件下很突出，反映了刺激的清晰开始（见 图 1D）。为了比较呼叫类型和噪声刺激之间的平均 ERP，我们应用了单因素方差分析 （ANOVA），将刺激作为 Cz 响应中的受试者间因子。我们发现刺激分别在刺激开始后 13-18 ms、28-36 ms 和 45-88 ms 对 C...

为了研究语言的进化，比较人类和非人类灵长类动物的大脑机制很重要。我们开发了一种无创测量清醒动物脑电图 （EEG） 的方法。它使我们能够直接长期比较人类和动物之间的脑电图数据，而不会伤害它们。

Vocal communication plays a crucial role in the social interactions of primates, particularly in survival and social organization. Humans have developed a ...

electroencephalography-measurements-awake-marmosets-listening-to

Research

JoVE Journal

Neuroscience

Electroencephalography Measurements in Awake Marmosets Listening to Conspecific Vocalizations 

310 Views.  Kyoto University. We would like to know how vocal information is represented in the brain of small primates which are rich in vocal communications. We would like to illustrate how the representation will be altered by psychiatric disorders such as schizophrenia in the future. We compared non-invasive EEGs across four primate species, and found that the latency of auditory cortex to respond to sounds is longer in humans than in other primates.This would have a significant impact on the perception of comp...

Video: Author Spotlight: Investigating Vocal Information Representation in Small Primates and Its Alteration by Psychiatric Disorders Using Noninvasive EEG