Nov 18, 2021

Public workspaceCitric Acid Water Restriction

DOI
dx.doi.org/10.17504/protocols.io.buw4nxgw
  • 1University of Edinburgh
Thomas R Clarke
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DOI: dx.doi.org/10.17504/protocols.io.buw4nxgw
Protocol Citation: Thomas R Clarke 2021. Citric Acid Water Restriction. protocols.io https://dx.doi.org/10.17504/protocols.io.buw4nxgw
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: May 11, 2021
Last Modified: November 18, 2021
Protocol Integer ID: 49852
Keywords: Mouse, Behaviour, Water Restriction, Citric Acid
Abstract
How to prepare and carry out citric acid water restriction for motivating mice to engage in behaviour.
Safety warnings
Any mice dropping to below 80% of their pre-CA baseline weight must be returned to ad lib water (see Duguid Lab PPL for regulations)
Baseline Weights
Baseline Weights
3d
3d
Handle animals and weigh for 3-5 days to establish a mean baseline weight prior to introduction of the CA water
Preparing 2% CA Water
Preparing 2% CA Water
15m
15m
Prepare the following:

  • 2 bottles
  • Citric acid crystals
  • Sterile Gloves or weighing boats
  • Scales
Empty one of the water bottles, place it on the scales and tare/zero
Pour the water from the other bottle into the one on the scales, record the weight of water
Calculate 2% weight of the water, this is the weight of citric acid crystals you need

e.g. 300g of water, requires 6g of citric acid (300*0.02 = 6)
Use the plastic side of the packet containing the sterile gloves or a weighing boat to weigh out the citric acid crystals
Add crystals to the bottle of water, put on the lid/spout and shake with finger covering the opening until dissolved
Label the bottle on 2 sides as "2% Citric Acid". Add a 'DO NOT REMOVE' label to the base of the bottle
Starting water restriction
Starting water restriction
5m
5m
Replace normal water bottle with 2% CA bottle, perform behaviour with 10% sucrose as a reward
Weigh animals daily to ensure they do not drop below 80% of their baseline weight


Plot from Urai 2021 (https://www.eneuro.org/content/8/1/ENEURO.0230-20.2020) showing typical progression of weight loss for mice during CA water restriction

If any animal drops below 80% of their baseline weight, return to ad lib water per the PPL