Automated pup-level analysis reveals distinct effects of prenatal CBD and THC exposure on maternal retrieval

This study examined how prenatal CBD and {Delta}9-tetrahydrocannabinol (THC) exposure affects maternal caregiving in C57BL/6J mice. We developed the Machine-Automated Scoring of the Pup Retrieval Test (MAS-PRT) to overcome limitations of manual behavioral scoring. MAS-PRT integrates Multi-Animal DeepLabCut for dam and pup tracking with Detectron2 for dynamic nest reconstruction. Validated against 170 manually annotated retrieval trials, the pipeline showed high concordance with manual measurements and enabled reproducible extraction of encounter latency, retrieval latency, and locomotor trajectories. Prenatal exposure did not impair general nest-building or overall home-cage maternal care. However, pups from both CBD and THC groups showed reduced body weight at postnatal day 5. Cox proportional hazards modeling revealed divergent effects by compound: CBD-exposed dams exhibited a weight-dependent increase in probability of encountering and retrieving lighter pups, independent of pup sex. Spatial tracking further showed that dams traversed significantly shorter total trajectories when retrieving female progeny exposed to either compound. Longitudinal trial-by-trial analysis indicated intact task acquisition in controls and CBD dams, whereas THC dams displayed a flattened learning curve driven by lower retrieval latencies on initial trials. Together, these findings indicate that prenatal cannabinoid exposure does not produce generalized disruption of maternal care but instead induces compound-specific alterations in maternal reactivity, retrieval kinematics, and learning dynamics.