这项研究得到了美国国立卫生研究院国家耳聋和其他沟通障碍研究所的F31DC017053-01A1（Lenell，PI）和K23DC014517（Johnson，PI）的资助。

本研究是根据纽约大学医学院机构动物护理和使用委员会进行的。
1. 解剖大鼠喉部
<ol><li>根据机构批准的协议对大鼠实施安乐死。将腹颈部从下颌骨剃到手洗，并用酒精拭子擦拭，以防止组织标本中的毛皮污染。</li>
	<li>在具有10倍放大倍率的解剖镜下，通过用手术刀创建中线颈部切口来切除整个喉部，直到气管暴露。</li>
	<li>将腹侧外侧喉肌分开，使用镊子和解剖剪刀或手术刀暴露喉部。</li>
	<li>将气管尾部切开到第三气管环，并在舌骨上做一个切口，用解剖剪刀切除整个喉部。</li>
	<li>使用显微切割工具（镊子、销和微剪刀）从喉部取出外源性喉组织（食道、甲状腺和外源性喉肌）。</li>
	<li>使用微剪刀，使用后腱肌之间的中线作为标志，将喉背侧的肛门一分为二。用微剪刀将喉部的侧壁固定以暴露声带，然后通过声带前部之间的甲状腺软骨中线将腹侧一分为二（图1）。 注意:此步骤可以是可选的;可以跳过它以保持喉部完整。喉部的两切允许通过分别使用同一喉部的右侧和左侧来进行多种免疫染色技术。</li>
	<li>在磷酸盐缓冲溶液（PBS）中冲洗每个半喉部约10秒，并用任务刮水器轻轻干燥，以减少冷冻过程中的冰晶形成。</li>
</ol>2. 固定和/或快速冷冻喉组织
注意:固定可能不适用于所有免疫染色方案。喉组织通常在解剖后立即被快速冷冻。跳过步骤2.1，无需固定即可快速冷冻喉部组织。
<ol><li>为了固定半喉，将组织置于装有4%甲醛的PBS离心管中，室温下在70rpm的轨道振荡器上1小时。将组织转移到干净的离心管中，并在PBS中冲洗3次20分钟。然后转移到干净的离心管中，并在4°C下浸没在20%蔗糖/ 5%甘油溶液中（约18小时或直到组织下沉）。 注意:甲醛是危险的，应与适当的个人防护设备一起在通风橱中使用。</li>
	<li>将所有半喉部均匀放置到充满最佳切割温度（OCT）化合物的冷冻模具中。对于半边，将声带内侧表面面向冷冻器底部的组织放置，并将声带的纵向面平行于冷冻器开口的下边缘。对于整个喉部，将后侧胪骨的组织朝向冷冻膜的底部，并且声带的纵向面平行于冷冻孔的下边缘。 注意:OCT化合物内一致的喉部方向对于大鼠声带的冷冻切除至关重要。一旦包埋并冷冻了半边，就必须解冻以改变其方向，从而引入多次解冻-冷冻循环导致组织损伤的风险。</li>
	<li>使用异戊烷（2-甲基丁烷）在被液氮包围的钢制烧杯中冷却的闪冻组织。 注:当烧杯的侧面和底部开始形成白色沉淀物时，异戊烷达到组织冷冻的最佳温度13。使用异戊烷是因为它具有比液氮更高的导热性，这有助于防止组织块在快速冷冻过程中开裂。有关OTC中冷冻组织的更详细说明，请参阅Kumar等人13。</li>
	<li>将每个模具包裹在预先标记的箔中，并放入单独的冷冻袋中以防止脱水，并立即储存在干冰上，直到转移到-80°C的冰箱中储存。</li>
</ol>3. 横截面上的冷冻切除术
<ol><li>将低温恒温器中的腔室温度设置为-20°C，该温度介于制造商手册推荐用于肌肉组织切片的温度范围（15-25°C）的中间。</li>
	<li>将低温恒温器截面厚度设置为 10 μm 厚截面。 注意:对于肌肉纤维横截面分析，10μm厚的切片是最佳的，可以对标记的肌肉纤维进行完整的染色和强大的成像强度，以进行纤维分型分析14，15，16。某些方案可能需要不同的切片厚度，具体取决于神经肌肉靶标。</li>
	<li>将组织转移到低温恒温器室，在低温恒温器标本盘（卡盘）上加入一层均匀的OCT化合物，并将嵌入的组织块放在标本盘上的OCT化合物的顶部。要获得用于甲状腺样动脉 （TA） 肌纤维分析的声带横截面，请将标本贴在卡盘上，使腹侧甲状腺软骨面向低温恒温器叶片，而类肾上腺软骨面向标本盘。 注意:重要的是要注意，这些地标在此阶段是不可见的，因为OCT化合物在冷冻时会变白和不透明。这种可见性的缺乏是为什么在闪光冻结阶段注意下颌的方向至关重要的原因。</li>
	<li>通过将标本头部推进100μm直到甲状腺软骨的腹侧部分出现来修剪OCT化合物。</li>
	<li>然后修剪并跟踪从甲状腺软骨开始的30μm切片，直到固有层，内侧TA肌肉和外侧TA肌肉暴露。 注意:从甲状腺软骨开始，每100μm一次，应跟踪并注意喉部标志，以确保切片的角度不倾斜。 图2 表示横截面上以10倍放大倍率下的两组喉部特征。</li>
	<li>一旦达到目标TA肌肉，在10μm处收集带正电荷的载玻片上的切片。</li>
	<li>将切片储存在4°C的PBS中以保持水分，直到它们准备好染色。 注意:固定组织可以在PBS中储存长达一周，具体取决于IHC靶标，而未固定的组织应立即处理。</li>
</ol>4. 纵平面冷冻切除术
<ol><li>将低温恒温器室再次设置为-20°C，将截面厚度更改为30μm。 注意:对于 NMJ 分析，30−60 μm 的组织厚度可用于捕获喉肌内的几个完整的 NMJ，而不会碎裂神经末梢或运动终板11，12，17。</li>
	<li>为了获得用于TA肌肉NMJ分析的纵向声带切片，请将标本贴在卡盘上，使会厌朝向低温恒温器叶片，气管腔朝下朝向标本盘。</li>
	<li>通过将标本头部向前推进100μm直到甲状腺软骨出现来修剪OCT化合物。</li>
	<li>修剪和跟踪从甲状腺发病到TA肌肉的内侧和外侧分裂暴露的30μm切片。 注意:建议在纵平面上使用五组喉部标志，以跟踪组织向目标TA肌的深度进展。 图3 显示了纵平面上10倍放大镜下的喉部特征。</li>
	<li>一旦达到目标TA肌肉，在30μm处收集带正电荷的载玻片上的切片。</li>
	<li>将切片储存在4°C的PBS中以保持水分，直到它们准备好染色。</li>
</ol>

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作者没有什么可透露的。

准备用于神经肌肉分析的大鼠声带可能会带来各种挑战。喉部肌肉不仅小且被软骨包围，从而难以直接提取目标肌肉，而且在喉部解剖学标志物的深度上还发现了动物之间的高变异性。对于肌肉横截面方案，在腹侧甲状腺软骨最初出现后，完整的声带切片出现在21-85个切片（每节10μm）之间，这比NMJ分析方案纵平面中的63-126个截面（每节35μm）要少得多（表1）。
?...

大鼠喉是一个成熟的模型，用于研究结构和功能神经肌肉喉对发育，衰老，疾病和药理药物的适应性1，2，3，4，5。组织学方法的一致性对于这项工作至关重要，因为肌肉制备和分析涉及多种复杂性，以及与喉软骨内封装的肌肉的喉部大小，形状和形貌相关的挑战1，6，7，8，9，10，11.由于大鼠内在喉部肌肉的尺寸很小，系统地嵌入，冷冻和冷冻切除对于获得一致和准确的结果至关重要。例如，当在冠状平面上切割大鼠声带时，四个内在喉肌的神经肌肉接头（NMJ）位于组织深度小于1.8 mm以内11。因此，在冷冻切除过程中精确监测喉部肌肉解剖结构对于准确识别感兴趣的部分并防止组织过度切除至关重要。目标肌肉的过度切除可能导致 NMJs11 的数量和形貌识别不准确，或者如果目标肌肉由于特征性方向混淆而被丢弃，则可能导致样本量总体减少12。随着喉部肌肉研究及其各自适应性的新模型的开发，标准操作程序对于确保结果在研究中精确，可靠和可重复至关重要。
本文的目的是详细准备大鼠声带，以实现最佳纵向和横截面分析。描述了我们实验室中经常使用的详细方法，以在冷冻切除过程中识别目标肌肉标志物。虽然在几个实验室中使用了类似的方法，但这里提供了比文献中更多的细节，以确保新手研究人员实施时可靠和准确的复制。本教程的目标是为大鼠声带的免疫组织化学（IHC）评估提供标准方法，以提高实验室和研究的一致性。

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preparation of the rat vocal fold for neuromuscular analyses

本教程的目的是描述用于组织化学神经肌肉研究的大鼠声带的准备工作。该协议概述了大鼠喉部解剖，闪光冷冻和声带冷冻切除术的程序。本研究描述了如何在纵向和横截面上冷冻切除声带。该协议的新颖之处在于冷冻切除过程中的喉部跟踪，可确保准确识别内在喉部肌肉并减少组织丢失的机会。数字显示了两个平面的渐进式冷冻切除。对29只大鼠半喉部进行了冷冻切除术，并从甲状腺软骨的出现到包括整个声带的第一部分的外观进行了跟踪。两个平面上所有动物的完整声带都可视化了。从甲状腺软骨的外观到两个平面上的完整声带的距离存在很高的变异性。体重与喉部标志物的深度无关，这表明个体变异性以及与组织准备相关的其他因素可能是切片过程中标志物外观高变异性的原因。本研究详细介绍了一种方法，并提供了用于准备大鼠声带以进行组织化学神经肌肉检查的形态学数据。由于个体变异性高，在冷冻切除过程中应密切跟踪喉部标志物，以防止过度切除组织和组织丢失。使用一致的方法，包括足够的组织准备和对大鼠喉内标志物的认识，将有助于在研究中获得一致的结果，并帮助有兴趣使用大鼠声带作为模型的新研究人员研究喉神经肌肉机制。

代表性的结果是正在进行的声乐运动对喉神经肌肉系统影响的研究的一部分。对29只雄性Fischer 344/棕色挪威大鼠（12只9个月大，17只24个月大）进行称重和安乐死，吸入CO2 ，然后进行双侧开胸术。
这些程序遵循概述的协议来标记外侧和内侧TA肌肉的NMJ和纤维大小。使用喉部肌肉和周围软骨在纵向和横截面中跟踪喉标志物之间的距离，以确定冷冻切除术期间的进展（<stron...

Watch this Scientific Journal Video about Preparation of the Rat Vocal Fold for Neuromuscular Analyses at JoVE.com

Preparation of the Rat Vocal Fold for Neuromuscular Analyses

该协议描述了用于准备大鼠声带以进行组织化学神经肌肉研究的方法。

用于神经肌肉分析的大鼠声带的制备

The purpose of this tutorial is to describe the preparation of the rat vocal fold for histochemical neuromuscular study. This protocol outlines procedures ...

preparation-of-the-rat-vocal-fold-for-neuromuscular-analyses

Research

JoVE Journal

Neuroscience

2.6K 次观看.  New York University School of Medicine. 该协议描述了用于准备大鼠声带以进行组织化学神经肌肉研究的方法。

视频: Preparation of the Rat Vocal Fold for Neuromuscular Analyses

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 ...

科研

神经科学

Targeted Training of Ultrasonic Vocalizations in Aged and Parkinsonian Rats

Voice deficits are a common complication of both Parkinson disease (PD) and aging; they can significantly diminish quality of life by impacting communication abilities. 1, 2 Targeted training (speech/voice therapy) can improve specific voice deficits,3, 4 although the underlying mechanisms of behavioral interventions are not well understood. Systematic investigation of voice deficits and therapy should consider many factors that are difficult to control in humans, such as age, home environment, age post-onset of disease, severity of disease, and medications. The method presented here uses an animal model of vocalization that allows for systematic study of how underlying sensorimotor mechanisms change with targeted voice training. The ultrasonic recording and analysis procedures outlined in this protocol are applicable to any investigation of rodent ultrasonic vocalizations.
The ultrasonic vocalizations of rodents are emerging as a valuable model to investigate the neural substrates of behavior.5-8 Both rodent and human vocalizations carry semiotic value and are produced by modifying an egressive airflow with a laryngeal constriction.9, 10 Thus, rodent vocalizations may be a useful model to study voice deficits in a sensorimotor context. Further, rat models allow us to study the neurobiological underpinnings of recovery from deficits with targeted training.
To model PD we use Long-Evans rats (Charles River Laboratories International, Inc.) and induce parkinsonism by a unilateral infusion of 7 &mu;g of 6-hydroxydopamine (6-OHDA) into the medial forebrain bundle which causes moderate to severe degeneration of presynaptic striatal neurons (for details see Ciucci, 2010).11, 12 For our aging model we use the Fischer 344/Brown Norway F1 (National Institute on Aging).
Our primary method for eliciting vocalizations is to expose sexually-experienced male rats to sexually receptive female rats. When the male becomes interested in the female, the female is removed and the male continues to vocalize. By rewarding complex vocalizations with food or water, both the number of complex vocalizations and the rate of vocalizations can be increased (Figure 1).
An ultrasonic microphone mounted above the male's home cage records the vocalizations. Recording begins after the female rat is removed to isolate the male calls. Vocalizations can be viewed in real time for training or recorded and analyzed offline. By recording and acoustically analyzing vocalizations before and after vocal training, the effects of disease and restoration of normal function with training can be assessed. This model also allows us to relate the observed behavioral (vocal) improvements to changes in the brain and neuromuscular system.

Voice disorders are debilitating in aging and Parkinson disease. The ultrasonic vocalizations of rats, also affected by these conditions, can be used to study these voice disorders, their neural substrates, and the nature of functional recovery with behavioral intervention.

老年帕金森病大鼠的超声波发声的有针对性的培训

Voice deficits are a common complication of both Parkinson disease (PD) and aging; they can significantly diminish quality of life by impacting ...

Subcutaneous Neurotrophin 4 Infusion Using Osmotic Pumps or Direct Muscular Injection Enhances Aging Rat Laryngeal Muscles

Laryngeal dysfunction in the elderly is a major cause of disability, from voice disorders to dysphagia and loss of airway protective reflexes. Few, if any, therapies exist that target age-related laryngeal muscle dysfunction. Neurotrophins are involved in muscle innervation and differentiation of neuromuscular junctions (NMJs). It is thought that neurotrophins enhance neuromuscular transmission by increasing neurotransmitter release. The neuromuscular junctions (NMJs) become smaller and less abundant in aging rat laryngeal muscles, with evidence of functional denervation. We explored the effects of NTF4 for future clinical use as a therapeutic to improve function in aging human laryngeal muscles. Here, we provide the detailed protocol for systemic application and direct injection of NTF4 to investigate the ability of aging rat laryngeal muscle to remodel in response to NTF4 application. In this method, rats either received NTF4 either systemically via osmotic pump or by direct injection through the vocal folds. Laryngeal muscles were then dissected and used for histological examination of morphology and age-related denervation.

Here, we present a protocol to describe the use of neurotrophin 4 (NTF4) systemically and directly to remodel rat aging laryngeal muscles.

使用渗透泵或直接肌肉注射的皮下神经营养因子4输液增强老化大鼠喉肌肉

Laryngeal dysfunction in the elderly is a major cause of disability, from voice disorders to dysphagia and loss of airway protective reflexes. Few, if ...

发育生物学

Transplantation of Olfactory Ensheathing Cells to Evaluate Functional Recovery after Peripheral Nerve Injury

Olfactory ensheathing cells (OECs) are neural crest cells which allow growth and regrowth of the primary olfactory neurons. Indeed, the primary olfactory system is characterized by its ability to give rise to new neurons even in adult animals. This particular ability is partly due to the presence of OECs which create a favorable microenvironment for neurogenesis. This property of OECs has been used for cellular transplantation such as in spinal cord injury models. Although the peripheral nervous system has a greater capacity to regenerate after nerve injury than the central nervous system, complete sections induce misrouting during axonal regrowth in particular after facial of laryngeal nerve transection. Specifically, full sectioning of the recurrent laryngeal nerve (RLN) induces aberrant axonal regrowth resulting in synkinesis of the vocal cords. In this specific model, we showed that OECs transplantation efficiently increases axonal regrowth.
OECs are constituted of several subpopulations present in both the olfactory mucosa (OM-OECs) and the olfactory bulbs (OB-OECs). We present here a model of cellular transplantation based on the use of these different subpopulations of OECs in a RLN injury model. Using this paradigm, primary cultures of OB-OECs and OM-OECs were transplanted in Matrigel after section and anastomosis of the RLN. Two months after surgery, we evaluated transplanted animals by complementary analyses based on videolaryngoscopy, electromyography (EMG), and histological studies. First, videolaryngoscopy allowed us to evaluate laryngeal functions, in particular muscular cocontractions phenomena. Then, EMG analyses demonstrated richness and synchronization of muscular activities. Finally, histological studies based on toluidine blue staining allowed the quantification of the number and profile of myelinated fibers.
All together, we describe here how to isolate, culture, identify and transplant OECs from OM and OB after RLN section-anastomosis and how to evaluate and analyze the efficiency of these transplanted cells on axonal regrowth and laryngeal functions.

Olfactory ensheathing cells (OECs) are neural crest cells which allow growth of the primary olfactory neurons. This specific property can be used for cellular transplantation. We present here a model of cellular transplantation based on the use of OECs in a laryngeal nerve injury model.

嗅鞘细胞移植周围神经损伤后，以评估功能恢复

Olfactory ensheathing cells (OECs) are neural crest cells which allow growth and regrowth of the primary olfactory neurons. Indeed, the primary olfactory ...

A Murine Model of Cervical Spinal Cord Injury to Study Post-lesional Respiratory Neuroplasticity

A cervical spinal cord injury induces permanent paralysis, and often leads to respiratory distress. To date, no efficient therapeutics have been developed to improve/ameliorate the respiratory failure following high cervical spinal cord injury (SCI). Here we propose a murine pre-clinical model of high SCI at the cervical 2 (C2) metameric level to study diverse post-lesional respiratory neuroplasticity. The technique consists of a surgical partial injury at the C2 level, which will induce a hemiparalysis of the diaphragm due to a deafferentation of the phrenic motoneurons from the respiratory centers located in the brainstem. The contralateral side of the injury remains intact and allows the animal recovery. Unlike other SCIs which affect the locomotor function (at the thoracic and lumbar level), the respiratory function does not require animal motivation and the quantification of the deficit/recovery can be easily performed (diaphragm and phrenic nerve recordings, whole body ventilation). This pre-clinical C2 SCI model is a powerful, useful, and reliable pre-clinical model to study various respiratory and non-respiratory neuroplasticity events at different levels (molecular to physiology) and to test diverse putative therapeutic strategies which might improve the respiration in SCI patients.

Respiratory failure is the leading cause of death following a cervical spinal cord injury. Having a reproducible, quantifiable, and reliable pre-clinical animal model of respiratory failure induced by a partial cervical injury will help to understand the subsequent respiratory and non-respiratory neuroplasticity and allow testing putative repair strategies.

颈脊髓损伤的小鼠模型研究后皮损呼吸神经可塑性

A cervical spinal cord injury induces permanent paralysis, and often leads to respiratory distress. To date, no efficient therapeutics have been developed ...

医学

多层硅胶声带模型的 3D 打印

Manufacturing Process for Non-Adhesive Super-Soft Vocal Fold Models

This study aims to develop super-soft, non-sticky vocal fold models for voice research. The conventional manufacturing process of silicone-based vocal fold models results in models with undesirable properties, such as stickiness and reproducibility issues. Those vocal fold models are prone to rapid aging, leading to poor comparability across different measurements. In this study, we propose a modification to the manufacturing process by changing the order of layering the silicone material, which leads to the production of non-sticky and highly consistent vocal fold models. We also compare a model produced using this method with a conventionally manufactured vocal fold model that is adversely affected by its sticky surface. We detail the manufacturing process and characterize the properties of the models for potential applications. The outcomes of the study demonstrate the efficacy of the modified fabrication method, highlighting the superior qualities of our non-sticky vocal fold models.&#160;The findings contribute to the development of realistic and reliable vocal fold models for research and clinical applications.

作者聚焦：用于语音和机器人应用的合成声带模型的制造进展

This study demonstrates the manufacturing of non-sticky and super-soft vocal fold models by introducing a specific way to create the vocal fold layers, providing a detailed description of the manufacturing procedure, and characterizing the properties of the models.

非粘性超软人声折叠模型的制造工艺

This study aims to develop super-soft, non-sticky vocal fold models for voice research. The conventional manufacturing process of silicone-based vocal ...

工程学

In Vivo Evaluation of the Mechanical and Viscoelastic Properties of the Rat Tongue

The tongue is a highly innervated and vascularized muscle hydrostat on the floor of the mouth of most vertebrates. Its primary functions include supporting mastication and deglutition, as well as taste-sensing and phonetics. Accordingly, the strength and volume of the tongue can impact the ability of vertebrates to accomplish basic activities such as feeding, communicating, and breathing. Human patients with sleep apnea have enlarged tongues, characterized by reduced muscle tone and increased intramuscular fat that can be visualized and quantified by magnetic resonance imaging (MRI). The abilities to measure force generation and viscoelastic properties of the tongue constitute important tools for obtaining functional information to correlate with imaging data. Here, we present techniques for measuring tongue force production in anesthetized Zucker rats via electrical stimulation of the hypoglossal nerves and for determining the viscoelastic properties of the tongue by applying passive Lissajous force/deformation curves.

In Vivo Evaluation of the Mechanical and Viscoelastic Properties of the Rat Tongue

We describe a surgical procedure in an anesthetized rat model for determining the muscle tone and viscoelastic properties of the tongue. The procedure involves specific stimulation of the hypoglossal nerves and application of passive Lissajous force/deformation curves to the muscle.

大鼠舌的机械和粘弹性的体内评估

The tongue is a highly innervated and vascularized muscle hydrostat on the floor of the mouth of most vertebrates. Its primary functions include ...

Endoscopic Approach for Studying Laryngeal Dysfunction in Rodents

Minimally Invasive Murine Laryngoscopy for Close&#45;Up Imaging of Laryngeal Motion During Breathing and Swallowing

The larynx is an essential organ in mammals with three primary functions - breathing, swallowing, and vocalizing. A wide range of disorders are known to impair laryngeal function, which results in difficulty&#160;breathing (dyspnea), swallowing impairment (dysphagia), and/or voice impairment (dysphonia). Dysphagia, in particular, can lead to aspiration pneumonia and associated morbidity, recurrent hospitalization, and early mortality. Despite these serious consequences, existing treatments for laryngeal dysfunction are largely aimed at surgical and behavioral interventions that unfortunately do not typically restore normal laryngeal function, thus highlighting the urgent need for innovative solutions.
To bridge this gap, we have been developing an experimental endoscopic approach to investigate laryngeal dysfunction in murine (i.e., mouse and rat) models. However, endoscopy in rodents is quite challenging due to their small size relative to current endoscope technology, anatomical differences in the upper airway, and the necessity for anesthesia to optimally access the larynx. Here, we describe a novel transoral laryngoscopy approach that permits close-up, unobstructed video imaging of laryngeal motion in mice and rats. Critical steps in the protocol include precise anesthesia management (to prevent overdosing that abolishes swallowing and/or risks respiratory distress-related mortality) and micromanipulator control of the endoscope (for stable video recording of laryngeal motion by a single researcher for subsequent quantification).
Importantly, the protocol can be performed over time in the same animals to study the impact of various pathological conditions specifically on laryngeal function. A novel advantage of this protocol is the ability to visualize airway protection during swallowing, which is not possible in humans due to epiglottic inversion over the laryngeal inlet that obstructs the glottis from view. Rodents therefore provide a unique opportunity to specifically investigate the mechanisms of normal versus pathological laryngeal airway protection for the ultimate purpose of discovering treatments to effectively restore normal laryngeal function.

This protocol describes a serial transoral laryngoscopy approach for mice and rats that permits close-up, unobstructed video imaging of the larynx during breathing and swallowing using an optimized anesthetic regimen and finely tuned endoscopic manipulation techniques.

The larynx is an essential organ in mammals with three primary functions - breathing, swallowing, and vocalizing. A wide range of disorders are known to ...

用于神经肌肉分析的大鼠声带的制备

摘要

探索更多视频

此视频中的章节

由运动神经元调制生理，形态学和神经化学的表征

老年帕金森病大鼠的超声波发声的有针对性的培训

使用渗透泵或直接肌肉注射的皮下神经营养因子4输液增强老化大鼠喉肌肉

嗅鞘细胞移植周围神经损伤后，以评估功能恢复

颈脊髓损伤的小鼠模型研究后皮损呼吸神经可塑性

非粘性超软人声折叠模型的制造工艺

非粘性超软人声折叠模型的制造工艺

非粘性超软人声折叠模型的制造工艺

大鼠舌的机械和粘弹性的体内评估

Minimally Invasive Murine Laryngoscopy for Close-Up Imaging of Laryngeal Motion During Breathing and Swallowing