Research in the Kawakami lab was supported by NIH grants: R01 AR064418-01A1, R01 HL124283-01, R21 AI 115534-01, and R41AI124734-01.

All animal experiments were approved by the Animal Care and Use Committee of the La Jolla Institute for Allergy and Immunology.
1. Mouse Body Temperature Measurement During Anesthetization
<ol>
	<li>Place the mouse in an anesthesia induction box. Anesthetize by using 1 L/min flow of oxygen with 5% isoflurane. 
	NOTE: Anesthetization is confirmed when the mouse ceases voluntary movement and has been immobile for over 30 s. Alternatively, monitor respiratory rate, and once mice are breath...

<ol>
	<li>Finkelman, F. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anaphylaxis:+lessons+from+mouse+models.">Anaphylaxis: lessons from mouse models.</a> The Journal of Allergy and Clinical Immunology. 120 (3), 506-515 (2007).</li><li>Lee, J. K., Vadas, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anaphylaxis:+mechanisms+and+management.">Anaphylaxis: mechanisms and management.</a> Clinical and Experimental Allergy: Journal of the British Society for Allergy and Clinical Immunology. 41 (7), 923-938 (2011).</li><li>Newsom, D. M., Bolgos, G. L., Colby, L., Nemzek, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+body+surface+temperature+measurement+and+conventional+methods+for+measuring+temperature+in+the+mouse.">Comparison of body surface temperature measurement and conventional methods for measuring temperature in the mouse.</a> Contemporary Topics in Laboratory Animal Science. 43 (5), 13-18 (2004).</li><li>Wong, J. P., Saravolac, E. G., Clement, J. G., Nagata, L. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+a+murine+hypothermia+model+for+study+of+respiratory+tract+influenza+virus+infection.">Development of a murine hypothermia model for study of respiratory tract influenza virus infection.</a> Laboratory Animal Science. 47 (2), 143-147 (1997).</li><li>Quimby, J. M., Olea-Popelka, F., Lappin, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+digital+rectal+and+microchip+transponder+thermometry+in+cats.">Comparison of digital rectal and microchip transponder thermometry in cats.</a> Journal of American Association for Laboratory Animal Science. 48 (4), 402-404 (2009).</li><li>Mei, J., Riedel, N., Grittner, U., Endres, M., Banneke, S., Emmrich, J. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Body+temperature+measurement+in+mice+during+acute+illness:+implantable+temperature+transponder+versus+surface+infrared+thermometry.">Body temperature measurement in mice during acute illness: implantable temperature transponder versus surface infrared thermometry.</a> Scientific Reports. 8 (1), 3526 (2018).</li><li>Doyle, E., Trosien, J., Metz, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protocols+for+the+induction+and+evaluation+of+systemic+anaphylaxis+in+mice.">Protocols for the induction and evaluation of systemic anaphylaxis in mice.</a> Methods in Molecular Biology. 1032, 133-138 (2013).</li><li>Brandt, E. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mast+cells+are+required+for+experimental+oral+allergen-induced+diarrhea.">Mast cells are required for experimental oral allergen-induced diarrhea.</a> The Journal of Clinical Investigations. 112 (11), 1666-1677 (2003).</li><li>Ando, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Histamine-releasing+factor+enhances+food+allergy.">Histamine-releasing factor enhances food allergy.</a> The Journal of Clinical Investigations. 127 (12), 4541-4553 (2017).</li><li>Brandt, E. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeting+IL-4/IL-13+signaling+to+alleviate+oral+allergen-induced+diarrhea.">Targeting IL-4/IL-13 signaling to alleviate oral allergen-induced diarrhea.</a> The Journal of Allergy and Clinical Immunology. 123 (1), 53-58 (2009).</li></ol>

The authors have nothing to disclose.

The protocol described was established with the goal of measuring body temperature without the use of anesthesia. Despite its relative ease with which temperature readings can be obtained, there are several caveats that accommodate this technique, in addition to the more obvious effects such as handling stress and different ambient temperatures.
First, in order to maintain consistent temperature readings throughout the experiment, the location where the temperature is being measured must be pr...

Measurement of body temperature has been an essential part of monitoring the effects of anaphylactic symptoms in animal models1,2. Temperature differences have been traditionally measured by rectal probe thermometers in mice3,4. With these measurements, investigators have reliably portrayed differences in temperature among variables; however, this method is a time-consuming procedure and causes distress to mice, which can increase the body core temperature. Rectal probing can also cause mucosal tearing and infection3. Moreover, the mice should be anesthetized in order to humanely insert the rectal probe to measure the temperature3. This is a slow process, and it prohibits the measurement of successive temperatures within a short period of time. Furthermore, mice activity phenotypes cannot be observed during this time until the anesthetic is completely worn off, which is another time-consuming process. More recently, other reliable methods to measure body temperature have used subcutaneously implanted passive infrared transponder tags or radio transmitters that include a temperature sensor3,5,6. Although they are accepted as the ideal practice by some researchers, these methods are not widely used because of high initial costs and distress to mice, due to the surgical implantation of a temperature sensor under the skin or another part of the body.
In order to demonstrate that a temperature difference is an accurate reflection of symptoms in a disease model1,2, mice must be awake during the temperature measurement and be able to return to their normal phenotypic activity immediately before and after the measurement. To this end, we sought a method by which this could be achieved.
Our goal was to accurately and inexpensively measure mouse body temperature, without the need for anesthesia and without restrictions on activity, to enable observation of behavioral phenotypes during and after the time of temperature measurement. To achieve this goal, it was apparent that a technique less invasive than the standard rectal temperature probes was required. Infrared thermometers have been used for decades in clinical medicine, especially in pediatrics, to obtain accurate temperature readings. It has been an alternative method that has allowed clinicians to quickly and accurately obtain temperature measurements in infants and fussy children that are actively mobile. We implemented this same technique in mice and have developed a successful method to obtain temperatures without anesthesia. Importantly, we show that this method is capable of replicating the well-established passive systemic anaphylaxis results regarding temperature changes, while also being able to observe the activity of the mouse throughout the measurement. Furthermore, we use the same method to evaluate body temperatures of food-allergic mice, while simultaneously investigating other symptoms, to demonstrate that body temperature is indeed an accurate reflection of the activity level and overall phenotype of the mouse.

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	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:260px;">Non-contact infrared thermometer</td>
			<td style="width:164px;">SinoPie</td>
			<td style="width:127px;">DT-8861</td>
			<td style="width:193px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Anti-dinitrophenyl (DNP) IgE</td>
			<td>Sigma Aldrich</td>
			<td>D8406-.2MG</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PrecisionGlide 30G needle</td>
			<td>BD</td>
			<td>305128</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PrecisionGlide 26G needle&#160;</td>
			<td>BD</td>
			<td>305111</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">1 mL syringe</td>
			<td>BD</td>
			<td>309659</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">Dinitrophenyl - human serum albumin</td>
			<td>Biosearch Technologies</td>
			<td>D-5059-10</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ovalbumin from chicken egg white</td>
			<td>Sigma Aldrich</td>
			<td>A5503-50G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Imject Alum</td>
			<td>ThermoFisher Scientific</td>
			<td>77161</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Animal Feeding Needles, disposable</td>
			<td>Fisher Scientific</td>
			<td>01-208-87</td>
			<td></td>
		</tr>
	</tbody>
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mouse body temperature measurement using infrared thermometer during passive systemic anaphylaxis and food allergy evaluation

Mouse body temperature measurement is of paramount importance for investigating allergies and anaphylactic symptoms. Rectal probes for temperature readings is common, and they have been proven to be accurate and invaluable in this regard. However, this method of temperature measurement requires the mice to be anesthetized in order to insert the probe without injury to the animal. This limits the ability to observe other phenotypes of the mouse simultaneously. In order to investigate other phenotypes while measuring temperatures, rectal probes are not ideal, and another method is desired. Here, we introduce a noninvasive method of temperature measurement that foregoes the requirement for mouse anesthesia while maintaining equal reliability to rectal probes in measuring body temperature. We use an infrared thermometer that detects body surface temperatures at ranges between 2 and 150 mm. This method of body temperature measurement is successful in reliably replicating temperature change trends during passive system anaphylaxis experiments in mice. We show that body surface temperatures are about 2.0 &#176;C lower than rectal probe measurements, but the degree of temperature drop follows the same trend. Furthermore, we use the same technique to observe mice in a food allergy model to evaluate temperature and activity levels simultaneously.

Passive systemic anaphylaxis: For iv injection, 10 week old female BALB/c mice were anesthetized. Prior to the injection, we measured their body temperatures (Video 1) as described in step 1. Figure 1 shows the temperature trend of both populations after iv injection. The IgE-sensitized mouse showed a maximum temperature drop of 3.0 &#176;C at 20 min, while the PBS control mouse had a maximum drop of 1.1 &#176;C at 20 min7</...

Watch this Scientific Journal Video about Mouse Body Temperature Measurement Using Infrared Thermometer During Passive Systemic Anaphylaxis and Food Allergy Evaluation at JoVE.com

Mouse Body Temperature Measurement Using Infrared Thermometer During Passive Systemic Anaphylaxis and Food Allergy Evaluation

Here we present a new method to accurately measure body temperature differences in passive systemic anaphylaxis (PSA) and food allergy mouse models using an infrared thermometer. This procedure has been accurately duplicated in previous PSA results.

Mouse body temperature measurement is of paramount importance for investigating allergies and anaphylactic symptoms. Rectal probes for temperature ...

This methodology helps address key questions in the field of allergy and immunology, specifically those that are regarding phenotypes that may be associated with body temperature changes. The main advantages of this technique are that it is inexpensive, non-invasive, and does not require anesthesia. The implications of this technique allow researchers to reduce the stress and pain of mice during temperature measurement while eliminating the effects of anesthesia, and/or to visualize behavioral phenotypes.To measure the mouse body temperature after anesthesia, confirm sedation by respiratory rate or immobility and grasp the mouse by the nape of the neck with an index finger and thumb and the tail with a pinky finger. While holding the mouse parallel to the lab bench surface, place an infrared thermometer sensor below the lower abdomen with the outer flat surface of the thermometer two to five millimeters away from the surface of the abdomen between the two upper nipples. Then depress the trigger of the thermometer to measure the temperature of the animal, taking care to keep the mouse and the thermometer steady.To measure the mouse body temperature without anesthetic, pick the mouse up by the middle of the tail and expose the abdomen. Allow the mouse to hold on to a straight edge surface with its forepaws and hold the trigger to measure the temperature. Alternatively, allow the mouse to hold on to the upper straight edge of the thermometer and make the mouse sit on the outer flat surface of the thermometer with it's abdomen just over the infrared sensor during the temperature measurement.For a passive systemic anaphylaxis sensitization use a one milliliter syringe equipped with a 26 gauge needle to inject 200 microliters of freshly prepared Anti-Dinitrophenol, or DNP, IgE in PBS, into the peritoneum of each recipient animal just lateral to the midline between the most inferior and second most inferior nipples. 24 hours after injection, measure the body temperature of each recipient animal as demonstrated, and load a one milliliter syringe equipped with a 30 gauge needle with 100 microliters of freshly prepared DNP human serum albumin. To induce passive systemic anaphylaxis, insert the needle on the medial side of the eye at a shallow angle aiming behind the eye, and inject the entire volume of solution intravenously into the retro-orbital venous sinus.Then place the mouse alone in a recovery cage with monitoring until full recumbency, and measure the body temperature and activity every 10 minutes for 70 minutes. IgE sensitized mice demonstrate a maximum temperature drop of three degrees Celsius at 20 minutes while PBS control mice exhibit a maximum drop of 1.1 degrees Celsius within the same time period after challenge. Behavioral observations can also be performed in non-anesthetized animals to assess the relationship between the observed temperature drops and the mouse activity.In this representative mouse food-allergy challenge, the PBS control group demonstrated a significant temperature drop 10 minutes after challenge compared to the 2CA inhibitor treated group. Evaluation of the activity scores and temperature drops from this food allergy challenge revealed a statistically significant correlation between the two sets of data. Following this procedure, other experiments involving mouse body temperature measurements can be performed to answer additional questions regarding behavioral phenotypes that are associated with body temperature change.

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Research

JoVE Journal

Immunology and Infection

25.1K vues.  La Jolla Institute for Allergy and Immunology. Cette méthodologie aide à répondre aux questions clés dans le domaine de l’allergie et de l’immunologie, en particulier celles qui sont concernant les phénotypes qui peuvent être associés à des changements de température corporelle. Les principaux avantages de cette technique sont qu’elle est peu coûteuse, non invasive, et ne nécessite pas d’anesthésie. Les implications de cette technique permettent aux chercheurs de réduire le stress et la douleur des souris lors de la m...