Introduction: Stereotaxic ablation is widely used in animal models to create precise lesions in specific brain regions. This approach allows for a detailed study of the functions of the lesioned areas and their behavioral, physiological, and neurochemical implications. Objective: In this study, bilateral ablation of the nucleus accumbens (NAcc) was performed in Wistar rats to investigate the behavioral and neurophysiological effects of functional removal of this region. Methods: Surgery was conducted using the EFF-331 stereotaxic device, with anatomical coordinates based on the Paxinos and Franklin atlas. The lesion was made using radiofrequency applied via a 4 mm needle developed specifically for this procedure, under general anesthesia combined with local anesthesia. Following the procedure, the animals were monitored for 28 days and received appropriate analgesia to ensure postoperative well-being. Results and discussion: The results indicated that the technique enabled precise and localized lesions in the NAcc, with minimal interference with adjacent structures, ensuring reproducibility and strict control of experimental variables. Behavioral analysis demonstrated significant alterations related to NAcc functions, contributing to the understanding of the neurobiological mechanisms involved in motivated behaviors and emotional states. The study highlights the importance of rigorous ethical and methodological care in the use of animal models and recommends complementing behavioral data with histological and neurochemical analyses to confirm the extent and specificity of the lesions, especially considering individual anatomical variations and post-lesion neuroplastic responses. Conclusion: the bilateral ablation of the NAcc via stereotaxy proved to be a safe, effective, and reproducible procedure with great potential for neurofunctional investigations and advancing the understanding of neural circuits related to reward, motivation, and emotional regulation.