Abstract
All mammalian mothers form some sort of caring bond with their infants that is crucial to the development of their offspring. The Pup Retrieval Test (PRT) is the leading procedure to assess pup-directed maternal care in laboratory rodents, used in a wide range of basic and preclinical research applications. Most PRT protocols require manual scoring, which is prone to bias and spatial and temporal inaccuracies. This study proposes a novel procedure using machine learning algorithms to enable reliable assessment of PRT performance. Automated tracking of a dam and one pup was established in DeepLabCut and was combined with automated behavioral classification of “maternal approach”, “carrying” and “digging” in Simple Behavioral Analysis (SimBA). Our automated procedure estimated retrieval success with an accuracy of 86.7%, whereas accuracies of “approach”, “carry” and “digging” were estimated at respectively 99.3%, 98.6% and 85.0%. We provide an open-source, step-by-step protocol for automated PRT assessment, which aims to increase reproducibility and reliability, and can be easily shared and distributed.
Highlights
All mammalian mothers form some sort of caring bond with their infants that is crucial to the development of their offspring
C57BL6J mice were subjected to the Pup Retrieval Test (PRT) on postnatal day 5 (P5)
Pup retrieval is an essential feature of maternal care in mice, and PRT performance has been used to study parental behavior in various research applications
Summary
All mammalian mothers form some sort of caring bond with their infants that is crucial to the development of their offspring. The Pup Retrieval Test (PRT) is the leading procedure to assess pupdirected maternal care in laboratory rodents, used in a wide range of basic and preclinical research applications. Some authors resorted to limited end-point registration to simplify manual PRT s coring[27,28] Components such as “maternal latency to start retrieving the pup”, “time necessary to complete retrieval”, and “efficacy of the retrieval” have been scored during real-time observation or from video recordings, but pup retrieval is a dynamic, interactive behavior that contains more information than these variables[28].
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