这项研究是由美国国立卫生研究院（批准1UM1HL112856）的支持。

所有的程序和协议批准了伊利诺伊大学芝加哥分校的机构动物护理和使用委员会。这里描述的实验是急性非存活实验中，也没有使用眼软膏。无菌条件下维持，只有当外科手术器械在DIH 2 O洗涤，用70％的乙醇发生大鼠在实验结束时牺牲经由第IV氯胺酮/赛拉嗪的药物过量发生。 1.准备仪器和化学品<ol><li>制备0.05M的5-羟色胺的盐酸在PBS原液。然后稀释股票与PBS到最终5-HT的0.203毫米的浓度。 </li><li>稀屈大麻酚在芝麻油在20微克/微升的浓度。 </li><li>切聚乙烯20厘米长（PE）的-50油管（ID0.58毫米，外径0.965毫米）。在管道的一端，切割用剪刀斜面尖端，而在另一方端，插入一个23克针。此23克针连接到1毫升注射器，并填写了0.3毫升50 U / ml肝素。 </li><li>切成四片4-0编织丝线，并设置到侧。 </li><li>消毒手术剪，2弯曲格雷夫钳，并用70％乙醇3微夹钳在DIH 2 O. </li></ol> 2.导管股静脉协议导管从耶斯佩森等进行修改。21 <ol><li>麻醉一个的Sprague-Dawley大鼠腹膜内（IP）的氯胺酮/赛拉嗪（100毫克/公斤10毫克/公斤）。捏的大鼠的脚趾和观察到任何运动，以确认麻醉的一个适当的水平。剃左大腿的腹侧部分，而颈的腹面。如果由本地机构动物护理和使用委员会必要，应用眼膏，以防止干燥。 </li><li>安全大鼠在手术板仰卧位置。用手术剪剪去皮肤左后肢大腿。 </li><li>使用镊子，行为钝dissection个肤浅的肌肉暴露股静脉。使用镊子到静脉从股动脉中分离出来，并将其放置2个线程围绕股静脉。 </li><li>使用弯曲格雷夫钳拉静脉阻止血液流动。用22克注射器针头，穿刺股静脉，然后插入PE-50管插入静脉的倾斜端。 </li><li>检查是否在PE-50管材是由回缩注射器的柱塞正确插入。血液应被视为进入的PE-50管材。配合周围的静脉和PE-50管2节使用2个线程。 </li></ol> 3.通过静脉输注5-HT引起的呼吸暂停<ol><li>周围放置老鼠的压电应变计来测量呼吸。 </li><li>用于经由放大器软件的电子放大器在计算机上的放大电平改变设置：扩增（100X）和带通滤波器（1-10赫兹）从所述应变仪获得的呼吸信号中，设置“highpas滤波器”在＆＃8220; AC @ 1 HZ“和”低通滤波器“，在”10赫兹“，并通过输入设定放大”10“为”初始增益“和”100“的”总收益“。 </li><li>由通过录音软件的计算机上打开“模拟输入”更改设置采样率。以使用模拟 - 数字转换器，“样品”在“1000赫兹”和数字化（500赫兹的采样速率）“跳过”由“1”（有效抽样率500赫兹）每隔一个的记录点，然后记录该信号使用刻录软件。 </li><li>取出1ml注射器从导管和插入一个500μl的精密玻璃注射器填充有0.203毫5-HT的进入导管。 </li><li>放置500微升精密玻璃注射器插入输液泵。注入12.5微克/千克每350微升/ kg的5-羟色胺的溶液在63毫升/小时的速度。执行多个输液，观察呼吸暂停大鼠，这被看作一个SA暂停在计算机显示器上所记录的呼吸呼吸信号（≥2.5秒）。 </li><li>在继续进行颈部手术，监测呼吸模式，并检查疼痛反射从脚趾捏在大鼠。如呼吸不规则，或如果从脚趾捏痛反射，管理IP氯胺酮/甲苯​​噻嗪（100毫克/公斤：5毫克/千克），然后再确定麻醉的适当水平。 </li></ol> 4.头颈外科杂志揭露节状神经节<ol><li>安全大鼠在手术板仰卧位置。使用手术剪在颈部正中纵向切。 </li><li>在颈阔肌（使用2微夹具保持这一肌肉清晰的手术部位的）使用钝性分离，露出胸骨舌骨肌和omohyoideus肌肉。分开这些肌肉，露出颈内动脉及其分支之一，翼腭动脉。 </li><li>观察的迷走神经，因为它沿内颈动脉运行，然后沿第<html> Ë翼腭动脉。观察一下迷走神经和翼腭动脉，随着舌咽神经（颅神经IX）与副神经（脑神经XI），进入后撕裂孔在颅底。 </li><li>注意，结状神经节被显示为迷走神经在进入后撕裂孔权之前的膨胀。还要注意，咽和喉上神经分支来的结状神经节的断前部和后部方面，分别为5 </li><li>使用格雷夫镊子，从动脉分开的迷走神经，并将其放置一块螺纹围绕迷走神经。放置在线程上的夹具上应用迷走神经轻微张力，和清洁任何结缔组织的结状神经节注入时提供较小的阻力。 </li><li>重复5-HT引起的呼吸暂停在第3节规定，确认无损伤做是为了结状神经节。 </li></ol>书名“&gt; 5。Intranodose神经节注射屈大麻酚<ol><li>补贴有特制28克半英寸的注射器针头斜面尖端5微升屈大麻酚的香油35°10微升气密精密玻璃注射器。 </li><li>放置在线程上的微型钳施加于迷走神经张力轻微，并刺破结状神经节，注意不要刺穿它。压下注射器缓慢（≥60秒）注入注射器的全部内容。注意，某些在注射器内容将漏出的神经节。 </li><li>在第3节规定，重复5-HT引起的呼吸暂停。 </li></ol>

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The authors declare that they have no competing financial interests.

的关键步骤成功注射神经化学物质的进节状神经节是：1）识别和清洁结缔组织断节状神经节; 2）确认注射前的结状神经节的完整性; 3）和使用小型量规针以微妙注入，但不能完全通过删截，所述结状神经节。 迷走神经支配许多器官在颈部和腹部，并传递重要的信息，如心脏速率，血压，支气管发炎，和胃肠膨胀到中枢神经系统。迷走神经的结状神经节包含受体氨基酸，单胺?...

Afferent signaling via the vagus nerve (cranial nerve X) transmits important general visceral information to the central nervous system (CNS) from baro-, chemo-, hepatic osmo-, cardiac, pulmonary, and gastric receptors located in the organs of the abdomen and thorax. The vagus nerve communicates information from stimuli such as heart rate, blood pressure, bronchopulmonary irritation, and gastrointestinal distension to the nucleus of solitary tract (NTS) of the medulla. The cell bodies of the pseudounipolar neurons of the vagus nerve are located in the nodose and petrosal ganglia, of which the majority is found in the former. Nodose ganglion cells contain a wealth of receptors for amino acids, monoamines, neuropeptides, and other neurochemicals that when activated, can modify afferent vagus nerve activity.1 Numerous innervations of the afferent vagus nerves coupled with the diversity of receptors located on the nodose ganglia illustrate the biological importance of this cranial nerve, and systemic drugs that do not cross into the CNS targeted at receptors on nodose ganglia can be used to treat various diseases, such as sleep apnea, gastroesophageal reflux disease, or chronic cough.2-4
The ease of access to the nodose ganglia lends itself to experimental manipulation by midline longitudinal incision made at the neck. The vagus nerve emerges from the posterior lacerated foramen at the base of the skull, and immediately displays a swelling of the nerve that is the nodose ganglion. The nodose ganglion is easily recognizable due to two nerve branches that arise from it: anteriorly the pharyngeal branch; and posteriorly superior laryngeal branch.5 Previous experimental manipulations of the nodose ganglia involved electrophysiological recordings,6 injections of immunohistochemical or immunofluorescent compounds for nerve tracings,7-10 superfusion or injections of neuroexcitotoxins,11-13 injections of adeno-associated virus to knockdown receptors,14,15 and injections of receptor-specific neurochemicals to change the activity of the vagus nerve.16,17
Systemic injections of neurochemicals are problematic in that systemic treatment affects both peripheral and central nervous systems. Thus, systemic treatment does not isolate the effect of neurochemicals on afferent vagal nerve activity. This protocol describes a method using readily available equipment of intranodose injections in the Sprague-Dawley rat that modulates vagus nerve activity without affecting the central nervous system. Stimulation of serotonin type 3 (5-HT3) receptors on nodose ganglia by intravenous (IV) infusion of serotonin (5-HT) induces the Bezold-Jarisch reflex, a vagal response trifecta of bradycardia, hypotension, and apnea, which can be abolished by supranodose vagatomy.11,17-19 Apnea is easily measured by placing a respiratory transducer around the abdomen of the rat.17,18 Cannabinoids decrease 5-HT-induced current in nodose ganglia cells,20 and intranodose ganglia injections of dronabinol attenuate 5-HT-induced apnea.

<table border="0" cellpadding="0" cellspacing="0" width="660">
	<tbody>
		<tr height="53">
			<td height="53" style="height:53px;width:191px;">Name of Material/ Equipment</td>
			<td style="width:155px;">Company</td>
			<td style="width:141px;">Catalog Number</td>
			<td style="width:175px;">Comments/Description</td>
		</tr>
		<tr height="53">
			<td height="53" style="height:53px;width:191px;">5-HT HCl</td>
			<td style="width:155px;">MP Biomedicals</td>
			<td style="width:141px;">215376591</td>
			<td style="width:175px;">12.5 &#181;g/kg per 350 &#181;l/kg</td>
		</tr>
		<tr height="53">
			<td height="53" style="height:53px;width:191px;">Dronabinol (Marinol) 10 mg Capsules (80 &#181;g/&#181;l)</td>
			<td style="width:155px;">AbbVie</td>
			<td style="width:141px;">NDC 0051-0023-21</td>
			<td style="width:175px;">Dilute with sesame oil to 20 &#181;g/&#181;l</td>
		</tr>
		<tr height="53">
			<td height="53" style="height:53px;width:191px;">Sesame Oil</td>
			<td style="width:155px;">Sigma-Aldrich</td>
			<td style="width:141px;">S3547</td>
			<td style="width:175px;"></td>
		</tr>
		<tr height="53">
			<td height="53" style="height:53px;width:191px;">Intramedic Polyethylene-50</td>
			<td style="width:155px;">BD</td>
			<td style="width:141px;">427411</td>
			<td style="width:175px;">Ordered from VWR (Cat. # 63019-047)</td>
		</tr>
		<tr height="53">
			<td height="53" style="height:53px;width:191px;">Graefe Forceps</td>
			<td style="width:155px;">Roboz</td>
			<td style="width:141px;">RS-5138</td>
			<td style="width:175px;">Two are needed</td>
		</tr>
		<tr height="53">
			<td height="53" style="height:53px;width:191px;">Johns Hopkins Bulldog Clamp</td>
			<td style="width:155px;">Roboz</td>
			<td style="width:141px;">RS-7441</td>
			<td style="width:175px;">Three are needed</td>
		</tr>
		<tr height="53">
			<td height="53" style="height:53px;width:191px;">Piezoelectric Strain Gauge</td>
			<td style="width:155px;">Ambu</td>
			<td style="width:141px;">813255-100</td>
			<td style="width:175px;"></td>
		</tr>
		<tr height="53">
			<td height="53" style="height:53px;width:191px;">Data Acquisition USB Subsystems</td>
			<td rowspan="2" style="width:155px;">DataWave Technologies</td>
			<td style="width:141px;">NA</td>
			<td style="width:175px;"></td>
		</tr>
		<tr height="53">
			<td height="53" style="height:53px;width:191px;">Sciworks Experimenter Software</td>
			<td style="width:141px;">NA</td>
			<td style="width:175px;"></td>
		</tr>
		<tr height="53">
			<td height="53" style="height:53px;width:191px;">CyberAmp</td>
			<td style="width:155px;">Axon Instruments</td>
			<td style="width:141px;">NA</td>
			<td style="width:175px;"></td>
		</tr>
		<tr height="53">
			<td height="53" style="height:53px;width:191px;">Syringe, 500 &#181;L, Model 1750 TLL</td>
			<td rowspan="3" style="width:155px;">Hamilton Company</td>
			<td style="width:141px;">81220</td>
			<td style="width:175px;"></td>
		</tr>
		<tr height="53">
			<td height="53" style="height:53px;width:191px;">Syringe, 10 &#181;L, Model 1801 RN</td>
			<td style="width:141px;">7659-01</td>
			<td style="width:175px;"></td>
		</tr>
		<tr height="53">
			<td height="53" style="height:53px;width:191px;">Needle, 28 gauge, Small Hub RN</td>
			<td style="width:141px;">7803-02</td>
			<td style="width:175px;">Point Style 4, Angle 35, Length 0.5 in</td>
		</tr>
	</tbody>
</table>

a method of nodose ganglia injection in sprague-dawley rat

Afferent signaling via the vagus nerve transmits important general visceral information to the central nervous system from many diverse receptors located in the organs of the abdomen and thorax. The vagus nerve communicates information from stimuli such as heart rate, blood pressure, bronchopulmonary irritation, and gastrointestinal distension to the nucleus of solitary tract of the medulla. The cell bodies of the vagus nerve are located in the nodose and petrosal ganglia, of which the majority are located in the former. The nodose ganglia contain a wealth of receptors for amino acids, monoamines, neuropeptides, and other neurochemicals that can modify afferent vagus nerve activity. Modifying vagal afferents through systemic peripheral drug treatments targeted at the receptors on nodose ganglia has the potential of treating diseases such as sleep apnea, gastroesophageal reflux disease, or chronic cough. The protocol here describes a method of injection neurochemicals directly into the nodose ganglion. Injecting neurochemicals directly into the nodose ganglia allows study of effects solely on cell bodies that modulate afferent nerve activity, and prevents the complication of involving the central nervous system as seen in systemic neurochemical treatment. Using readily available and inexpensive equipment, intranodose ganglia injections are easily done in anesthetized Sprague-Dawley rats.

图1表示在大鼠中有输注5-HT的前后intranodose神经节注射屈大麻酚诱导呼吸暂停呼吸样本记录。 5-HT的激活的5-HT 3受体上的结状神经节，有助于心动过缓，低血压和呼吸暂停的的Bezold-Jarisch反射。11,17-19 Intranodose神经节注射屈大麻酚的激活抑制性的CB受体，或变构调节5-HT的3受体抑制16,17,20,22的迷走神经。对5-HT诱导的激发前（ 图1，上图）和后?...

Watch this Scientific Journal Video about A Method of Nodose Ganglia Injection in Sprague-Dawley Rat at JoVE.com

A Method of Nodose Ganglia Injection in Sprague-Dawley Rat

Afferent vagal signaling transmits important information to central nervous system from receptors located in organs of the abdomen and thorax. The nodose ganglia of vagus nerves contain many types of receptors that modulate vagal activity. This protocol describes a method of local injections of neurochemicals into the nodose ganglia.

结状神经节注射液只SD大鼠的方法

Afferent signaling via the vagus nerve transmits important general visceral information to the central nervous system from many diverse receptors located ...

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18.4K 次观看.  University of Illinois at Chicago. Afferent vagal signaling transmits important information to central nervous system from receptors located in organs of the abdomen and thorax. The nodose ganglia of vagus nerves contain many types of receptors that modulate vagal activity. This protocol describes a method of local injections of neurochemicals into the nodose ganglia.