Summary

Метод Узловые ганглиев инъекций в Спрэг Dawley крыс

Published: November 25, 2014
doi:

Summary

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.

Abstract

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.

Introduction

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.

Protocol

Все процедуры и протоколы были одобрены уходу и использованию комитета Институциональная животных из Университета Иллинойса в Чикаго. Эксперименты, описанные здесь, являются острые эксперименты, не выживания, и не было никакой пользы глазной мази. Поддержание стерильных условиях про…

Representative Results

Рисунок 1 представляет запись дыхательный образец у крыс, имевших вливание 5-НТ, чтобы вызвать апноэ до и после intranodose ганглиев инъекций дронабинолом. 5-HT активирует 5-HT 3 рецепторы на нодозного ганглиев, которые способствуют Bezold-Jarisch рефлекса брадикардия, гипотония и апноэ….

Discussion

Критические шаги для успешного введения нейрохимических минуте нодозного ганглиев являются: 1) выявление и очистки соединительной ткани и выключение нодозного ганглиев; 2) подтверждения целостности нодозного ганглиев перед инъекцией; 3) и с помощью небольшой иглы, чтобы впрыснуть в то?…

Divulgaciones

The authors have nothing to disclose.

Acknowledgements

Это исследование было поддержано Национальным институтом здравоохранения (грант 1UM1HL112856).

Materials

Name of Material/ Equipment Company Catalog Number Comments/Description
5-HT HCl MP Biomedicals 215376591 12.5 µg/kg per 350 µl/kg
Dronabinol (Marinol) 10 mg Capsules (80 µg/µl) AbbVie NDC 0051-0023-21 Dilute with sesame oil to 20 µg/µl
Sesame Oil Sigma-Aldrich S3547
Intramedic Polyethylene-50 BD 427411 Ordered from VWR (Cat. # 63019-047)
Graefe Forceps Roboz RS-5138 Two are needed
Johns Hopkins Bulldog Clamp Roboz RS-7441 Three are needed
Piezoelectric Strain Gauge Ambu 813255-100
Data Acquisition USB Subsystems DataWave Technologies NA
Sciworks Experimenter Software NA
CyberAmp Axon Instruments NA
Syringe, 500 µL, Model 1750 TLL Hamilton Company 81220
Syringe, 10 µL, Model 1801 RN 7659-01
Needle, 28 gauge, Small Hub RN 7803-02 Point Style 4, Angle 35, Length 0.5 in

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Citar este artículo
Calik, M. W., Radulovacki, M., Carley, D. W. A Method of Nodose Ganglia Injection in Sprague-Dawley Rat. J. Vis. Exp. (93), e52233, doi:10.3791/52233 (2014).

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