Name: Establishment of a Mouse Model with Cough Hypersensitivity via Inhalation of Citric Acid (Video) | JoVE Journal
Uploaded: 2025-01-10T13:40:00.000+00:00
Duration: 5 min 43 s
Description: Learn to establish a mouse model with cough hypersensitivity via citric acid inhalation for chronic cough research. This protocol enables mechanistic studies and therapeutic development using whole body plethysmography.

ai_agent_IP

ai_agent

maintenance

instantTrialDurationMinutes

isBiopharma

enableNewMyJoveSectionDesign

educationFeatures

seoMetaApi

ai_copilot_institution_blacklist

ai_chat_institution_blacklist

freeAccessWindow

ssoRoleCapture

scheduledMaintenanceBanner

joveLabsFeatures

disable_autoplay_for_bots

extended_trial_duration

5 minutes

This study was supported by the National Natural Science Foundation of China (NSFC 82100034), Guangzhou Science and Technology Planning Project (202102010168).

All animal experiment procedures were approved by the Laboratory Animal Ethics Committee of the First Affiliated Hospital of Guangzhou Medical University (20230656). Adult male specific-pathogen-free C57BL/6 mice, aged 8-10 weeks and weighing 20-25 g, were used in this study. The details of the reagents and equipment used are listed in the&#160;Table of Materials.
1. Chemical reagent preparation
<ol>
	<li>Citric acid (CA) solution preparation
	<ol>
		<li>For cough assessment, prepare a 0.4 M solution by dissolving 3.84 g of citric acid powder in normal saline to a final volume of 50 mL.</li>
		<li>For animal treatment, prepare a 0.1 M solution by dissolving 9.6 g of citric acid powder in normal saline to a final volume of 500 mL.</li>
	</ol>
	</li>
	<li>Capsaicin solution preparation
	<ol>
		<li>Dissolve 30.5 mg of capsaicin powder in PBS containing 10% ethanol and 10% Tween-80 to make a 10 mM stock solution.</li>
		<li>Dilute the stock solution 1:100 in normal saline to make a 100 &#181;M working solution.</li>
	</ol>
	</li>
	<li>Methacholine solution preparation
	<ol>
		<li>Dissolve 100 mg of methacholine powder in PBS to a final volume of 2 mL to make a 50 mg/mL solution.</li>
		<li>Dilute this solution with PBS to obtain concentrations of 25 mg/mL, 12.5 mg/mL, 6.25 mg/mL, and 3.125 mg/mL.</li>
	</ol>
	</li>
</ol>
2. Animal preparation
<ol>
	<li>Housing conditions
	<ol>
		<li>House mice in cages with free access to food and water, maintained at a temperature of 22 &#177; 1 &#176;C with a standard 12-h light/dark cycle. Experimental manipulations were performed after 1 week of acclimation to the feeding environment.</li>
	</ol>
	</li>
	<li>Acclimation
	<ol>
		<li>Allow mice to acclimate to the feeding environment for 1 week before conducting experimental manipulations.</li>
	</ol>
	</li>
</ol>
3. Development of the model
<ol>
	<li>Group assignment
	<ol>
		<li>Randomly divide the mice into a model group and a control group, with 8 mice in each group.</li>
	</ol>
	</li>
	<li>Exposure setup
	<ol>
		<li>Place the mice in two independent chambers (as shown in&#160;Figure 1), each connected to an ultrasonic nebulizer.</li>
		<li>Expose the model group to 0.1 M citric acid (CA) aerosol and the control group to 0.9% normal saline (NS) aerosol at an atomization rate of 3 mL/min for 2 h per day over 2 weeks (as shown in Figure 2). Return the mice to their cages after each exposure.</li>
	</ol>
	</li>
	<li>Cough sensitivity assessment
	<ol>
		<li>After the last exposure, assess cough sensitivity by evaluating reflexive cough (challenged with NS, CA (0.4 M), and capsaicin (100 &#181;M)) and spontaneous cough (without any challenge). 
		NOTE: To monitor dynamic changes in cough sensitivity during model development, assess cough sensitivity on days 0, 3, 7, and 10 using a CA (0.4 M) challenge.</li>
	</ol>
	</li>
</ol>
4. Cough sensitivity assessment
<ol>
	<li>Instrument preparation
	<ol>
		<li>Use a non-invasive whole-body plethysmography (WBP) system to measure cough sensitivity. Connect the chambers, flow transducers, bias flow, and other components according to the application manual (see Table of Materials).</li>
		<li>Perform a calibration process to ensure accurate measurements by clicking on the Calibrate button. A status screen will display the progress, and once the status bar reaches 100%, the calibration is complete.</li>
	</ol>
	</li>
	<li>Cough detection
	<ol>
		<li>Create a cough study for mice and configure the parameters for 1 min of acclimation, 10 min of response time, and 10 min of chemical solution delivery.</li>
		<li>Place the conscious mice into individual chambers, ensuring there is only one mouse per chamber. 
		NOTE: Make sure the mouse is completely inside the chamber before closing the lid to avoid damaging its tail and digits.</li>
		<li>Add 1 mL of the chemical solutions (NS, CA, or capsaicin) to the nebulizer. 
		NOTE: The number of cough events-each consisting of three phases: inspiratory, compressive, and expulsive15 (as shown in Figure 3)-will be recorded once the study begins. After 10 min of recording, cough detection will be complete. Clean the chamber after detecting each animal to avoid cross-contamination.</li>
	</ol>
	</li>
</ol>
5. Airway Hyperresponsiveness (AHR) measurement
<ol>
	<li>Instrument preparation
	<ol>
		<li>Prepare instruments as described in step 4.1.</li>
	</ol>
	</li>
	<li>AHR measurement
	<ol>
		<li>Create a dose-response study for mice, configuring the measurement types, parameters, and task sequence: 1 min for acclimation, 30 s for methacholine solution delivery, 3 min for response time, and 1 min for recovery.</li>
		<li>Place the conscious mice into individual chambers, ensuring there is only one mouse per chamber.</li>
		<li>Add 50 &#181;L of the chemical solutions (PBS or methacholine) to the nebulizer and initiate the study.</li>
		<li>Record the value of Penh, an indicator of bronchoconstriction, in response to different concentrations of methacholine (0 mg/mL, 3.125 mg/mL, 6.25 mg/mL, 12.5 mg/mL, 25 mg/mL, 50 mg/mL).</li>
	</ol>
	</li>
</ol>
6. Bronchoalveolar lavage collection
<ol>
	<li>Sacrifice
	<ol>
		<li>Sacrifice the mice using hyperanesthesia with pentobarbital sodium (50 mg/kg) administered via intraperitoneal injection (following institutionally approved protocols).</li>
	</ol>
	</li>
	<li>Sample collection
	<ol>
		<li>Collect blood from the orbital sinus of the mouse into a 1.5 mL microcentrifuge tube, then place the tube on ice. Centrifuge the blood at 3000 x g for 10 min at 4&#176;C. Use a pipette to collect the supernatant, aliquot it, and store it at -80 &#176;C.</li>
		<li>Collect bronchoalveolar lavage fluid (BALF) by opening the chest to expose the trachea, inserting a 22 G indwelling needle into the trachea, then lavaging three times with 0.5 mL of pre-cooled PBS and collecting the fluid.</li>
		<li>After collection, centrifuge the BALF at 500 x g for 10 min at 4 &#176;C. Collect the supernatant, aliquot it, and store it at -80 &#176;C. Resuspend the pellet in 100 &#181;L of PBS and prepare cell smears for further analysis.</li>
		<li>Collect lung tissues by opening the chest to expose the lungs, removing the lung tissues, and placing them into cryogenic vials. Store the vials at -80 &#176;C for future studies.</li>
	</ol>
	</li>
</ol>
7. Quantitative RT-PCR
<ol>
	<li>Extract total RNA from lung tissue using TRIzol reagent. Perform cDNA synthesis using a First-strand cDNA synthesis kit, following the manufacturer&#39;s instructions (see Table of Materials).</li>
	<li>Conduct PCR in a real-time quantitative PCR detection system using SYBR green fluorescence. Normalize the quantification of relative SP and CGRP mRNA expression to the expression of &#946;-Actin9.</li>
</ol>
8. Statistical analysis
<ol>
	<li>Present data as the mean &#177; SEM. Use a t-test to compare data between two groups, considering P &#60; 0.05 as statistically significant. Conduct all statistical analyses using statistical and graphing software.</li>
</ol>

Citric Acid-Induced Mouse Model for Cough Hypersensitivity

<ol><li>Morice, A. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Epidemiology+of+cough%5BTitle%5D)+AND+%22Pulm+Pharmacol+Ther%22%5BJournal%5D)">Epidemiology of cough.</a> Pulm Pharmacol Ther. 15 (3), 253-259 (2002).</li>
<li>Lai, K., Long, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Current+status+and+future+directions+of+chronic+cough+in+China%5BTitle%5D)+AND+%22Lung%22%5BJournal%5D)">Current status and future directions of chronic cough in China.</a> Lung. 198 (1), 23-29 (2020).</li>
<li>Zeiger, R. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Patient-reported+burden+of+chronic+cough+in+a+managed+care+organization%5BTitle%5D)+AND+%22J+Allergy+Clin+Immunol+Pract%22%5BJournal%5D)">Patient-reported burden of chronic cough in a managed care organization.</a> J Allergy Clin Immunol Pract. 9 (4), 1624-1637.e10 (2021).</li>
<li>Chamberlain, S. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+impact+of+chronic+cough%3A+A+cross-sectional+European+survey%5BTitle%5D)+AND+%22Lung%22%5BJournal%5D)">The impact of chronic cough: A cross-sectional European survey.</a> Lung. 193 (3), 401-408 (2015).</li>
<li>Morice, A. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Expert+opinion+on+the+cough+hypersensitivity+syndrome+in+respiratory+medicine%5BTitle%5D)+AND+%22Eur+Respir+J%22%5BJournal%5D)">Expert opinion on the cough hypersensitivity syndrome in respiratory medicine.</a> Eur Respir J. 44 (5), 1132-1148 (2014).</li>
<li>Mazzone, S. B., Undem, B. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Vagal+afferent+innervation+of+the+airways+in+health+and+disease%5BTitle%5D)+AND+%22Physiol+Rev%22%5BJournal%5D)">Vagal afferent innervation of the airways in health and disease.</a> Physiol Rev. 96 (3), 975-1024 (2016).</li>
<li>Shapiro, C. O., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Airway+sensory+nerve+density+is+increased+in+chronic+cough%5BTitle%5D)+AND+%22Am+J+Respir+Crit+Care+Med%22%5BJournal%5D)">Airway sensory nerve density is increased in chronic cough.</a> Am J Respir Crit Care Med. 203 (3), 348-355 (2021).</li>
<li>Ando, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Neural+correlates+of+cough+hypersensitivity+in+humans%3A+Evidence+for+central+sensitization+and+dysfunctional+inhibitory+control%5BTitle%5D)+AND+%22Thorax%22%5BJournal%5D)">Neural correlates of cough hypersensitivity in humans: Evidence for central sensitization and dysfunctional inhibitory control.</a> Thorax. 71 (4), 323-329 (2016).</li>
<li>Plevkova, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Animal+models+of+cough%5BTitle%5D)+AND+%22Respir+Physiol+Neurobiol%22%5BJournal%5D)">Animal models of cough.</a> Respir Physiol Neurobiol. 290, 103656(2021).</li>
<li>Smith, J. A., Hilton, E. C. Y., Saulsberry, L., Canning, B. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Antitussive+effects+of+memantine+in+guinea+pigs%5BTitle%5D)+AND+%22Chest%22%5BJournal%5D)">Antitussive effects of memantine in guinea pigs.</a> Chest. 141 (4), 996-1002 (2012).</li>
<li>Hewitt, M. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Pharmacology+of+bradykinin-evoked+coughing+in+guinea+pigs%5BTitle%5D)+AND+%22J+Pharmacol+Exp+Ther%22%5BJournal%5D)">Pharmacology of bradykinin-evoked coughing in guinea pigs.</a> J Pharmacol Exp Ther. 357 (3), 620-628 (2016).</li>
<li>Zhong, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Antitussive+activity+of+the+Schisandra+chinensis+fruit+polysaccharide+%28SCFP-1%29+in+guinea+pigs+models%5BTitle%5D)+AND+%22J+Ethnopharmacol%22%5BJournal%5D)">Antitussive activity of the Schisandra chinensis fruit polysaccharide (SCFP-1) in guinea pigs models.</a> J Ethnopharmacol. 194, 378-385 (2016).</li>
<li>Zhong, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Effects+of+Schisandra+chinensis+extracts+on+cough+and+pulmonary+inflammation+in+a+cough+hypersensitivity+guinea+pig+model+induced+by+cigarette+smoke+exposure%5BTitle%5D)+AND+%22J+Ethnopharmacol%22%5BJournal%5D)">Effects of Schisandra chinensis extracts on cough and pulmonary inflammation in a cough hypersensitivity guinea pig model induced by cigarette smoke exposure.</a> J Ethnopharmacol. 165, 73-82 (2015).</li>
<li>Wei, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Effects+of+Shiwei+longdanhua+formula+on+LPS-induced+airway+mucus+hypersecretion%2C+cough+hypersensitivity%2C+oxidative+stress+and+pulmonary+inflammation%5BTitle%5D)+AND+%22Biomed+Pharmacother%22%5BJournal%5D)">Effects of Shiwei longdanhua formula on LPS-induced airway mucus hypersecretion, cough hypersensitivity, oxidative stress and pulmonary inflammation.</a> Biomed Pharmacother. 163, 114793(2023).</li>
<li>Chen, L., Lai, K., Lomask, J. M., Jiang, B., Zhong, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Detection+of+mouse+cough+based+on+sound+monitoring+and+respiratory+airflow+waveforms%5BTitle%5D)+AND+%22PLoS+One%22%5BJournal%5D)">Detection of mouse cough based on sound monitoring and respiratory airflow waveforms.</a> PLoS One. 8 (3), e59263(2013).</li>
<li>Gibson, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Treatment+of+unexplained+chronic+cough%3A+Chest+guideline+and+expert+panel+report%5BTitle%5D)+AND+%22Chest%22%5BJournal%5D)">Treatment of unexplained chronic cough: Chest guideline and expert panel report.</a> Chest. 149 (1), 27-44 (2016).</li>
<li>Nakaji, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Airway+remodeling+associated+with+cough+hypersensitivity+as+a+consequence+of+persistent+cough%3A+An+experimental+study%5BTitle%5D)+AND+%22Respir+Investig%22%5BJournal%5D)">Airway remodeling associated with cough hypersensitivity as a consequence of persistent cough: An experimental study.</a> Respir Investig. 54 (6), 419-427 (2016).</li>
<li>Xu, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Association+of+cough+hypersensitivity+with+tracheal+trpv1+activation+and+neurogenic+inflammation+in+a+novel+guinea+pig+model+of+citric+acid-induced+chronic+cough%5BTitle%5D)+AND+%22J+Int+Med+Res%22%5BJournal%5D)">Association of cough hypersensitivity with tracheal trpv1 activation and neurogenic inflammation in a novel guinea pig model of citric acid-induced chronic cough.</a> J Int Med Res. 46 (7), 2913-2924 (2018).</li>
<li>Iwata, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mechanical+stimulation+by+postnasal+drip+evokes+cough%5BTitle%5D)+AND+%22PLoS+One%22%5BJournal%5D)">Mechanical stimulation by postnasal drip evokes cough.</a> PLoS One. 10 (11), e0141823(2015).</li>
<li>Chen, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Establishment+of+a+mouse+model+with+all+four+clinical+features+of+eosinophilic+bronchitis%5BTitle%5D)+AND+%22Sci+Rep%22%5BJournal%5D)">Establishment of a mouse model with all four clinical features of eosinophilic bronchitis.</a> Sci Rep. 10 (1), 10557(2020).</li>
<li>Zhang, C., Lin, R. L., Hong, J., Khosravi, M., Lee, L. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Cough+and+expiration+reflexes+elicited+by+inhaled+irritant+gases+are+intensified+in+ovalbumin-sensitized+mice%5BTitle%5D)+AND+%22Am+J+Physiol+Regul+Integr+Comp+Physiol%22%5BJournal%5D)">Cough and expiration reflexes elicited by inhaled irritant gases are intensified in ovalbumin-sensitized mice.</a> Am J Physiol Regul Integr Comp Physiol. 312 (5), R718-R726 (2017).</li>
<li>Amato, A., Becci, A., Beolchini, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Citric+acid+bioproduction%3A+The+technological+innovation+change%5BTitle%5D)+AND+%22Crit+Rev+Biotechnol%22%5BJournal%5D)">Citric acid bioproduction: The technological innovation change.</a> Crit Rev Biotechnol. 40 (2), 199-212 (2020).</li>
<li>Hu, W., Li, W. J., Yang, H. Q., Chen, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Current+strategies+and+future+prospects+for+enhancing+microbial+production+of+citric+acid%5BTitle%5D)+AND+%22Appl+Microbiol+Biotechnol%22%5BJournal%5D)">Current strategies and future prospects for enhancing microbial production of citric acid.</a> Appl Microbiol Biotechnol. 103 (1), 201-209 (2019).</li>
<li>Morice, A. H., Kastelik, J. A., Thompson, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Cough+challenge+in+the+assessment+of+cough+reflex%5BTitle%5D)+AND+%22Br+J+Clin+Pharmacol%22%5BJournal%5D)">Cough challenge in the assessment of cough reflex.</a> Br J Clin Pharmacol. 52 (4), 365-375 (2001).</li>
</ol>

The authors have nothing to disclose.

This study successfully established a mouse model with cough hypersensitivity through continuous inhalation of citric acid (CA). This model demonstrated a reliable increase in cough sensitivity for both spontaneous coughs and reflexive coughs elicited by citric acid and capsaicin. Citric acid and capsaicin are widely used to assess cough reflex sensitivity16.
Several critical steps in this protocol ensure...

Cough is a crucial defensive reflex that helps clear respiratory secretions or foreign materials from the airway. However, it is also one of the most common symptoms of many respiratory diseases, often prompting patients to seek medical attention1. Chronic cough, defined as a persistent cough lasting more than 8 weeks in adults, significantly impacts the quality of life, causing issues such as incontinence, insomnia, reflux, and other unpleasant experiences, along with a substantial economic burden2,3,4. It is widely believed that increased cough sensitivity is a pathophysiological hallmark of chronic cough, where low levels of thermal, mechanical, and chemical irritants can trigger coughing5. Cough hypersensitivity is associated with airway inflammation6, remodeling of airway sensory nerves7, and alterations in the central nervous system8, though the precise molecular mechanisms remain unclear and require further elucidation through suitable animal models.
Various animals, including guinea pigs, cats, rabbits, dogs, and pigs, have been used to study the mechanisms of cough9. Guinea pigs have traditionally been recognized as the most suitable model for studying cough mechanisms and the efficacy of antitussive drugs9,10,11,12. However, these models have several experimental limitations, including high costs, a lack of transgenic tools, and a scarcity of commercial reagents. Additionally, guinea pigs often exhibit poor environmental tolerance and high mortality when exposed to stimuli. In contrast, mice are smaller, easier to maintain, more cost-effective, and amenable to genetic manipulation, making them more suitable for mechanistic investigations. Previous studies on cough models have primarily focused on cough induced by airway inflammation, mainly used to evaluate the efficacy of antitussive drugs and peripheral mechanisms13,14. There is currently a lack of animal models for cough hypersensitivity.
In response, we introduce a method for establishing a mouse model of cough hypersensitivity&#160;via&#160;continuous inhalation of citric acid (CA). This model is simpler, easier to construct, and more feasible compared to other animal models.

<table><tbody><tr><td>0.9% normal saline</td><td>Biosharp</td><td>BL158A</td><td></td></tr><tr><td>Capsaicin</td><td>Cayman chemical</td><td>92350</td><td></td></tr><tr><td>Citric Acid</td><td>Sigma-Aldrich</td><td>C2404</td><td></td></tr><tr><td>Ethanol</td><td>Guangzhou chemical reagent factory</td><td>GSHB15-AR-0.5L</td><td></td></tr><tr><td>First-strand cDNA synthesis kit</td><td>TransGen Biotech</td><td>AT341</td><td></td></tr><tr><td>Methacholine</td><td>Sigma-Aldrich</td><td>A2251</td><td></td></tr><tr><td>Non-invasive whole-body plethysmography (WBP) system</td><td>DSI</td><td>601-1400-001</td><td></td></tr><tr><td>Pentobarbital sodium</td><td>Merk</td><td>P3761</td><td></td></tr><tr><td>PerfectStart Green qPCR SuperMix</td><td>TransGen Biotech</td><td>AQ601</td><td></td></tr><tr><td>Phosphate Buffered Saline (PBS)</td><td>Meilunbio</td><td>MA0015</td><td></td></tr><tr><td>Real-time&amp;nbsp;quantitative&amp;nbsp;PCR&amp;nbsp;detecting&amp;nbsp;system</td><td>Bio-rad&amp;nbsp;</td><td>CFX&amp;nbsp; Connect</td><td></td></tr><tr><td>TRIzol reagent</td><td>Invitrogen</td><td>15596026CN</td><td></td></tr><tr><td>Tween-80</td><td>Solarbio</td><td>T8360-100</td><td></td></tr><tr><td>Ultrasonic nebulizer</td><td>Yuwell</td><td>402AI</td><td></td></tr></tbody></table>

establishment of a mouse model with cough hypersensitivity via inhalation of citric acid

Establishment of a Mouse Model with Cough Hypersensitivity via Inhalation of Citric Acid

Cough is one of the most common symptoms of many respiratory diseases. Chronic cough significantly impacts quality of life and imposes a considerable economic burden. Increased cough sensitivity is a pathophysiological hallmark of chronic cough. It has been observed that cough hypersensitivity is related to airway inflammation, remodeling of airway sensory nerves, and alterations in the central nervous system. However, the precise molecular mechanisms remain unclear and require further elucidation using suitable animal models. Previous studies have utilized guinea pigs as models for studying cough, but these models present several experimental limitations, including high costs, a lack of transgenic tools, and a scarcity of commercial reagents. In addition, guinea pigs typically exhibit poor environmental tolerance and high mortality when exposed to stimuli. In contrast, mice are smaller, easier to maintain, more cost-effective, and amenable to genetic manipulation, making them more suitable for mechanistic investigations. In this study, we established a mouse model with cough hypersensitivity&#160;via&#160;continuous inhalation of citric acid (CA). This model is straightforward to operate and yields reproducible results, making it a valuable tool for further studies on the mechanisms and potential novel treatments for chronic cough.

As shown in Figure 4A, cough sensitivity in the model group (CA group) significantly increased after 1 week of exposure compared to the control group (NS group), and this heightened sensitivity persisted throughout the exposure period. Neither the control group nor the model group mice experienced mortality during the modeling process (Figure 4B). Figure 4C and Figure 4D demonstrate that the number of spontaneous cough events si...

This protocol describes the development of a mouse model with cough hypersensitivity, which can serve as an ideal model for studying the mechanisms of chronic cough.

Cough is one of the most common symptoms of many respiratory diseases. Chronic cough significantly impacts quality of life and imposes a considerable ...

establishment-mouse-model-with-cough-hypersensitivity-via-inhalation

Research

JoVE Journal

Medicine

Experimental mouse models of allergic asthma offer new possibilities for studying disease pathogenesis and developing new therapeutics. These models are well suited to measuring factors governing the allergic immune response, airway inflammation, and pulmonary pathophysiology.

Murine Model of Allergen Induced Asthma

The present protocol provides a detailed step-by-step description of the procedures required to execute measurements of respiratory system mechanics as well as the assessment of airway responsiveness to inhaled methacholine in mice using the forced oscillation technique (flexiVent; SCIREQ Inc, Montreal, Qc, Canada).

Evaluation of Respiratory System Mechanics in Mice using the Forced Oscillation Technique

The goal of this protocol is to provide automated methods to quantify chronic lung pathologies in a murine model of COPD. The protocol includes exposing mice to cigarette smoke (CS), measuring pulmonary function, inflating the lungs, and using morphometry methods to measure emphysema and small airway remodeling in mice.

Automated Measurement of Pulmonary Emphysema and Small Airway Remodeling in Cigarette Smoke-exposed Mice

While in vitro study of host-pathogen interactions allow the characterization of specific immune responses, in vivo models are required to observe the effects of complex responses. Using Candida albicans exposure followed by Pseudomonas aeruginosa-mediated lung infection, we established a murine model of microbial interactions involved in ventilator-associated pneumonia pathogenicity.

Studying Microbial Communities In Vivo: A Model of Host-mediated Interaction Between Candida Albicans and Pseudomonas Aeruginosa in the Airways

This study describes the successful generation of a new chronic obstructive pulmonary disease (COPD) animal model by repeatedly exposing mice to high concentrations of ozone.

Generation of a Chronic Obstructive Pulmonary Disease Model in Mice by Repeated Ozone Exposure

Presented here is a useful protocol for lung fixation that creates a stable condition for histological evaluate of lung specimens from a mouse model of emphysema. The main advantage of this model is that it can fix many lungs with the same constant pressure without lung collapse or deflation.

Lung Fixation under Constant Pressure for Evaluation of Emphysema in Mice

This protocol describes a method for establishing a mouse model of silicosis through repeated exposure to silica suspensions via a nasal drip. This model can efficiently, conveniently, and flexibly mimic the pathological process of human silicosis with high repeatability and economy.

A Silicosis Mouse Model Established by Repeated Inhalation of Crystalline Silica Dust

This article presents a streamlined protocol for establishing a pulpitis model in mice using an innovative mouth-gag, followed by subsequent histological analysis.

Establishment of a Murine Pulp Exposure Model with a Novel Mouth-Gag for Pulpitis Research

Here, we describe the measurement of cough using a noninvasive and real-time whole-body plethysmography (WBP) system and the normative procedures for harvesting tissue samples of mice and introduce some methods to assess airway inflammation.

Methods for Detecting Cough and Airway Inflammation in Mice

This study established cough and sneeze models in mice by applying specific stimuli to the trachea and nasal cavity, and developed an audio-based method to distinguish between mouse coughs and sneezes by analyzing differences in their acoustic signals.<br...

Precision Induction and Distinction of Coughing and Sneezing Reflexes in Mice

Learn to establish a mouse model with cough hypersensitivity via citric acid inhalation for chronic cough research. This protocol enables mechanistic studies and therapeutic development using whole body plethysmography.

Establishment of a Mouse Model with Cough Hypersensitivity via Inhalation of Citric Acid

Explore More Videos