Mice homozygous for either the obese gene in linkage group XI or the diabetes gene in linkage VIII exhibit syndromes similar to those of diabetes mellitus in man, including obesity and hyperglycaemia. A study of the lipolysis in vitro of the epididymal adipose tissue of 5 genotypes of mice, namely, ob/ob, ob/ob+, ob+/ob+, db/db, and db+/db+, demonstrates that the mutant homozygotes respond differently from wild type homozygotes to various lipolytic agents. The agents tested were epinephrine, norepinephrine, isoproterenol, corticotropin 1-24, theophylline and N6, O2-dibutyryl-cyclic-adenosine3′,5′-monophosphate. The response of ob/ob+ tissue was the same as that of ob+/ob+ tissue. The data indicate that there is a defect with respect to lipolysis in ob/ob and db/db mice. This is discussed in light of the involvement of cyclic AMP. © 1969 Springer-Verlag.