Objective: Parasympathetic nervous system modulates adipose tissue as an endocrine gland. Botulinum toxin (BTX) may exert a lipolytic effect by interfering with acetylcholine transmission at the cholinergic parasympathetic nerve endings. Materials and Methods: Fifteen male rabbits were divided into three equal groups: one control group (A) and two case groups (B and C). The abdomens of all rabbits were divided into a 3-by-3-square grid. The control and two case groups received 9 subcutaneous injections of 0.9% normal saline, 1 U BTX (group B) and 2 U BTX (group C), respectively. Four weeks later the entire grid was excised from the abdominal area. Hematoxylin-eosin-stained tissue was used for stereological analysis to estimate cell surface and volume in 100 randomly selected cells. Results: Gross thinning of subcutaneous fat, shattering and disappearance of fat globules were seen in both case groups. Fat cell volume was reduced by 65% in group B (p=0.009) and by 77% in group C (p=0.009) compared to the control animals. Fat cell surface also decreased by 51% in group B (p=0.009) and 63% in the group C rabbits (p=0.009) compared to the control animals. Conclusion: Our pilot animal study revealed a dose-dependent lipolytic effect of subcutaneous BTX injection.