Name: a reversible, non-invasive method for airway resistance measurements and bronchoalveolar lavage fluid sampling in mice
Uploaded: 2010-04-13T17:00:00
Description: Watch this Scientific Journal Video about A Reversible, Non-invasive Method for Airway Resistance Measurements and Bronchoalveolar Lavage Fluid Sampling in Mice at JoVE.com

We thank Dr. W. Mintzer for the suggestion to perform fiberoptic orotracheal intubation. Supported by grants U19AI070973, R01AI057696, K02HL75243, and R01HL082487 from the National Institutes of Health.

Allergen challenge:
<ol>
	<li>C57BL/6 mice, 4-8 weeks of age, are anesthetized in an airtight plexiglass chamber purged with a 3.2% isoflurane in oxygen vapor mixture for 10 minutes to achieve deep general anesthesia.</li>
	<li>Intranasal allergen challenges (45&mu;L OVA (22.5 &mu;g) and 7&mu;L A. Oryzae (7 &mu;g), in PBS) are administered, every Tuesday, Thursday and Sunday, for a total of seven consecutive applications.</li>
</ol>
Anesthesia:
<ol start="3">
	<li>Prior to each allergen...

<ol>
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The study of asthma, and various other airway obstructive diseases, constitutes an active and expanding field of biomedical research. An important component of asthma-related experimental research is the capacity to measure changes in airway size under varying conditions. Excessive airway narrowing in response to provocative challenge, a canonical feature of asthma and related lung diseases and a property of the airway termed airway hyperresponsiveness, is a major component of clinically significant attacks leading to sh...

Airway physiology measurement software (Rescomp) was custom prepared (Millenium Premier Group; 415-519-4371). Data was analyzed using a PC workstation running Windows XP equipped with a Pentium III CPU (Intel, Inc. Santa Clara, CA) and a 17-pin analog to digital signal converter (National Instruments, #PC-LPM16). A small animal airway physiology workstation was custom assembled (Millenium Premier Group) using commercially available pressure transducers (part #TRD5700 and TRD4510), preamp modules (part #MAX2270), chassis (part # MAX1320; all from Buxco, Inc. Wilmington, NC) and a customized small animal plethysmograph. 0.5mm external diameter fiber-optic thread, connected to light source (Cole Palmer Illuminator, 41722 series) Ventilator (Harvard Apparatus Mouse Ventilator, #687) 10 mm, 27ga needle (BD Biosciences, cat. no. 309602) Heat lamp 1 ml syringe (BD Biosciences, cat. no. 305109) 4 clamps (Pony 3200 spring clamp) 0.5 mm external wire for intubation guide Hemacytometer Superfrost/plus microscope slides (Fisher cat. no. 12-550-15) Shandon Filter Cards (Thermo cat. no. 5991022) Differential cell slide stain (Fisher cat. no. 122911) Light microscope (Leica) Cytospin 3 (Shandon) 20 ga, 1.25 inch ProtectIV intravenous catheters (Smith Medical) 0.5 mm polymer optical fiber (Edmund Optics # NT02-532).

a reversible, non-invasive method for airway resistance measurements and bronchoalveolar lavage fluid sampling in mice

Airway hyperreactivity (AHR) measurements and bronchoalveolar lavage (BAL) fluid sampling are essential to experimental asthma models, but repeated procedures to obtain such measurements in the same animal are generally not feasible.  Here, we demonstrate protocols for obtaining from mice repeated measurements of AHR and bronchoalveolar lavage fluid samples.  Mice were challenged intranasally seven times over 14 days with a potent allergen or sham treated.  Prior to the initial challenge, and within 24 hours following each intranasal challenge, the same animals were anesthetized, orally intubated and mechanically ventilated.  AHR, assessed by comparing dose response curves of respiratory system resistance (RRS) induced by increasing intravenous doses of acetylcholine (Ach) chloride between sham and allergen-challenged animals, were determined.  Afterwards, and via the same intubation, the left lung was lavaged so that differential enumeration of airway cells could be performed.  These studies reveal that repeated measurements of AHR and BAL fluid collection are possible from the same animals and that maximal airway hyperresponsiveness and airway eosinophilia are achieved within 7-10 days of initiating allergen challenge.  This novel technique significantly reduces the number of mice required for longitudinal experimentation and is applicable to diverse rodent species, disease models and airway physiology instruments.  

Watch this Scientific Journal Video about A Reversible, Non-invasive Method for Airway Resistance Measurements and Bronchoalveolar Lavage Fluid Sampling in Mice at JoVE.com

A Reversible, Non-invasive Method for Airway Resistance Measurements and Bronchoalveolar Lavage Fluid Sampling in Mice

Repeated measurements of rodent respiratory physiology and sampling of airway inflammatory cells are desirable, but generally not feasible. Here we describe a repeatable method for orally intubating mice that permits repeated measurements of airway hyperreactivity and sampling of airway inflammatory cells.

Airway hyperreactivity (AHR) measurements and bronchoalveolar lavage (BAL) fluid sampling are essential to experimental asthma models, but repeated ...

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