Objective To evaluate the differential behavioral effects of intracerebral stereotaxic versus intramuscular injection of Botulinum Neurotoxin Serotype A (BoNT/A) in Spontaneously Hypertensive Rats (SHRs). Methods Thirty 4-week-old male SHRs were randomized into five groups (n=6): Model Control (MC), Stereotaxic Sham (IC-Sham), Stereotaxic BoNT/A (IC-BoNT/A), Intramuscular Vehicle (IM-Vehicle), and Intramuscular BoNT/A (IM-BoNT/A). Behavioral assessments included the Open Field Test and Morris Water Maze. Results In the Open Field Test, the IC-BoNT/A group exhibited significantly increased total distance traveled and average velocity compared to the MC group (P<0.05). Moreover, the hyperactivity observed in the IC-BoNT/A group was significantly more pronounced than in the IM-BoNT/A group (P<0.05). In the Morris Water Maze, the IC-BoNT/A group showed significantly prolonged escape latency, along with reduced platform crossings, distance traveled in the target quadrant, and time spent in the target quadrant compared to both the MC and IM-BoNT/A groups (P<0.05). Conclusions Compared to intramuscular injection, stereotaxic injection of BoNT/A significantly exacerbates hyperactivity, impulsivity, and cognitive deficits in SHRs. These findings suggest that the stereotaxic route more effectively blocks SNAP-25 function, providing a superior experimental approach for investigating the role of the SNARE complex in ADHD pathogenesis.