Longitudinal diffusion tensor imaging identifies coordinated white matter remodeling during cocaine self-administration in mice

White matter microstructural alterations are increasingly recognized as a feature of cocaine use disorder (CUD), yet the longitudinal progression and biological underpinnings of these changes remain poorly understood. Although rodent intravenous self-administration (IVSA) provides a translational model of voluntary cocaine use, longitudinal diffusion tensor imaging (DTI) has not previously been combined with cocaine IVSA in mice. Here, we established a longitudinal IVSA-DTI framework to characterize white matter microstructural remodeling during cocaine self-administration and early withdrawal. Male C57BL/6 mice underwent DTI at baseline, following 11 consecutive days of cocaine (0.5 mg/kg/infusion) or saline IVSA, and after a 7-day withdrawal period immediately following a cocaine-seeking test (n = 4/group). Fractional anisotropy (FA) was evaluated longitudinally using atlas-based region-of-interest analyses and whole-brain diffusion maps. Cocaine-treated mice acquired robust operant responding and exhibited coordinated longitudinal FA alterations across interconnected hippocampal-associated white matter pathways. The most consistent changes occurred within the bilateral dorsal hippocampal commissure, fornix, and fimbria, with cocaine-treated mice demonstrating increased FA during self-administration relative to baseline while saline controls exhibited smaller or opposing longitudinal changes. Difference-in-differences maps further demonstrated cocaine-associated FA alterations within hippocampal pathways and localized regions of the corpus callosum after accounting for longitudinal changes in saline controls. These findings establish the feasibility of combining longitudinal DTI with cocaine IVSA in mice and demonstrate coordinated white matter microstructural remodeling during voluntary cocaine exposure. This framework provides a translational platform for future studies integrating longitudinal neuroimaging with genetically modified mouse models, tractography, and histological validation to investigate the mechanisms underlying cocaine-associated white matter plasticity.