Dec 16, 2024

Public workspaceHypothermia Induction in Mice

DOI
dx.doi.org/10.17504/protocols.io.6qpvr937bvmk/v1
  • Mukesh Kumar1,
  • Catherine L. Pontius2,
  • Rose E. Witte3,
  • Kartik N. Rajagopalan2,
  • Timothy A. Ryan1,4
  • 1Department of Biochemistry, Weill Cornell Medicine, New York, NY 10065;
  • 2Department of Internal Medicine, UT Southwestern Medical Center, Dallas, TX 75390, USA;
  • 3Laboratory of Molecular Genetics, Howard Hughes Medical Institute, Rockefeller University, New York, NY 10065, USA;
  • 4Aligning Science Across Parkinson's (ASAP) Collaborative Research Network, Chevy Chase, Maryland 20815, USA
Mukesh Kumar
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DOI: dx.doi.org/10.17504/protocols.io.6qpvr937bvmk/v1
Protocol Citation: Mukesh Kumar, Catherine L. Pontius, Rose E. Witte, Kartik N. Rajagopalan, Timothy A. Ryan 2024. Hypothermia Induction in Mice. protocols.io https://dx.doi.org/10.17504/protocols.io.6qpvr937bvmk/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: November 26, 2024
Last Modified: December 16, 2024
Protocol Integer ID: 114382
Keywords: ASAPCRN
Funders Acknowledgements:
NIH
Grant ID: NS036942
NIH
Grant ID: NS11739
ASAP
Grant ID: ASAP-024404
KL2 Award by Rockefeller University
Abstract
This protocol describes a detailed method for inducing and measuring hypothermia in mice using Etomoxir and KLH45, enabling the evaluation of pharmacological effects on thermoregulation.
Guidelines
The protocol needs prior approval by the users' Institutional Animal Care and Use Committee (IACUC) or equivalent ethics committee.
Materials
Materials:

  • ReagentEtomoxirMerck MilliporeSigma (Sigma-Aldrich)Catalog #236020 : 30 mg/kg, dissolved in phosphate-buffered saline (PBS).
  • ReagentKLH45Merck MilliporeSigma (Sigma-Aldrich)Catalog #SML1998 : 40 mg/kg, dissolved in emulphor (18:1:1 saline: ethanol: ReagentPolyethylene Glycol Monooleyl EtherFisher ScientificCatalog #P071325G
  • Control vehicles: PBS or emulphor.

Equipments:

  • Heating block or water bath for solution preparation
  • Syringes and needles for intraperitoneal (IP) injection
  • Rectal thermometer for mice (Kent Scientific, RET-3)
  • Cage with controlled environment for post-injection monitoring

Preparation of Solutions
Preparation of Solutions
Dissolve Etomoxir in PBS to achieve the desired concentration for 30 mg/kg dosing.

Prepare KLH45 in emulphor composed of 18:1:1 saline: ethanol: polyethylene glycol monooleyl ether to achieve a 40 mg/kg dosing concentration.

Prepare control vehicles (PBS for Etomoxir and emulphor for KLH45).

Pre-Injection Procedure
Pre-Injection Procedure
Place mice on food restriction 3 hours prior to the injection to minimize metabolic variability.

Weigh each mouse to calculate the appropriate injection volume based on body weight.

Hypothermia Induction
Hypothermia Induction
Administer the following intraperitoneally (IP):

Etomoxir group: 30 mg/kg in PBS.

KLH45 group: 40 mg/kg in emulphor.

Control groups: Corresponding vehicle (PBS or emulphor).

Ensure proper injection technique to avoid discomfort or improper dosing.

Body Temperature Measurement
Body Temperature Measurement
Measure the baseline body temperature (Tb) of each mouse using a rectal probe.

Record body temperature at the following time points post-injection: 0 hour (immediately after injection), 0.5 hour, 1 hour, and 3 hours.

Post-Injection Monitoring
Post-Injection Monitoring
Return the mice to their cages immediately after injection.
Ensure the mice are housed without access to feed during the post-injection monitoring period.
Monitor for any signs of distress or abnormal behavior during the experiment.
Data Recording
Data Recording
Record all temperature measurements and note any significant changes or observations during the experiment.

Analyze temperature data to assess the hypothermic response induced by the treatments.

Note
  • Ensure accurate dosing by calibrating the syringe and needle prior to injection.
  • Use sterile techniques when preparing solutions and administering injections.
  • Mice should be handled gently to minimize stress, which can influence body temperature measurements.
  • Include adequate sample size and control groups for statistical reliability.