A Standardized Protocol for Quantifying the Behavioral Dynamics of Food-seeking in Mice

Food is generally hidden in a natural environment and require free-living animals to search for it. Although such food-seeking behaviors involve motivation and exploration, previous studies examined food-seeking simply by measuring the time spent in the food zone or the frequency of pursuing food-cued context. Moreover, after discovering food, animals need to taste and smell it in order to evaluate their nutritional value or possible toxicity. However, researchers could not easily distinguish food-seeking from food-evaluating behaviors because food was visible or accessible throughout each test. Herein, we describe a behavioral protocol that triggers animals to show the behavioral dynamics of food-seeking (e.g., navigation, nose-digging, and paw-digging) and that exclusively elicits food-seeking without provoking any other food-evaluating behaviors. First, we prepared an open-field box with the floor covered with bedding. After we hid foods under the bedding of each corner, the test mice were habituated in this arena for four days (pre-test phase). On the next day (test phase), they were placed under the same conditions, but the foods previously hidden were removed. This process enabled the mice to perceive their surroundings as a food-hidden environment, which induced the animal to exhibit sustained food-seeking. In conclusion, the protocol presented here is a powerful method for provoking multiple forms of food-seeking and quantifies food-seeking independently from other food-related behavioral stages. ### Competing Interest Statement The authors have declared no competing interest.