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The β3-Adrenergic receptor (β3AR) and leptin are molecules involved in the regulation of energy balance. Recently, a mutation in the β3AR gene (Trp64Arg) has been reported to be associated with features of insulin resistance, weight gain and early onset of Type 2 Diabetes Mellitus. The aim of this study was to determine the frequency and clinical characteristics of the Trp64Arg mutation in the β3AR gene in Type 2 diabetic patients, its relationship with leptin levels, and its role in microangiopathic complications.187 Type 2 diabetic patients and 100 unrelated non-diabetic subjects were studied. There was no difference between the diabetic and nondiabetic subjects in the frequency of the Trp64 and Arg64 alleles (92.5% vs. 92.3% and 7.5% vs. 7.7%, respectively). Type 2 diabetic patients were divided into two groups according to the presence (n = 27) or absence of the mutation (n = 160).Mutation of the β3AR gene was not associated with any differences either in the clinical and metabolic parameters or microangiopathic complications. Type 2 diabetic patients carrying the Arg64 allele tended to have a lower diabetes duration, but this was not statistically significant. Plasma leptin levels were not different according to the β3AR genotype.The Trp64Arg mutation is not a major determinant of Type 2 diabetes and its microangiopathic complications. Moreover, this mutation was not clinically relevant in leptin regulation.