Social behavior of Peromyscus maniculatus rufinus and P. polionotus leucocephalus was observed in mixed groups of three mice each to determine the degree of ethological isolation and to observe be- havior patterns which might contribute to the isolation noted between the two species. Ten categories of behavior were selected as indicators of ethological isolation, and the amount of interspecific and intraspecific contacts observed are presented. Behavioral patterns of the two species are similar. Mice of each species directed most of the sexual behavior to animals of the same species; however, there was less restriction of some other types of social behavior to mice of the same species. It is estimated the ethological isolation would prevent interbreeding between the two species if they were to become sympatric. INTRODUCTION Ethological isolating mechanisms are usually the most important in restricting interbreeding between closely related animal species (Mayr, 1963), including species of the genus of cricetid rodents, Peromyscus. Closely related species of Peromyscus may be crossed in the laboratory, but in areas of sympatry the natural populations fail to interbreed due to ecological and ethological isolating mechanisms (Dice, 1940). Peromyscus maniculatus and P. polionotus are classi- fied in the same species group (Osgood, 1909), and although they have completely allopatric distributions, there is evidence that they would fail to interbreed if natural populations ever became sympatric. Laboratory crosses between the two species result in some fertile ofi- spring (Liu, 1953); however, Blair and Howard (1944), found evi- dence of ethological isolation between them. Ethological isolating mechanisms are based on restriction of the direction of sexual behavior by an animal to another animal that provides the specific stimuli characteristic of that species. Reinforcement could strengthen iso- lating mechanisms by narrowing the range of stimuli to which an animal will respond, or by resulting in avoidance of stimuli associated with another sympatric species. However, reinforcement is not neces- sary for the development of ethological isolation between two species (Godfrey, 1958). Observations of the behavior of mixed groups of mice were made in the laboratory to determine the degr ee of ethological isolation between the two species and to observe particular behavior sequences or differences in behavior that might contribute to the ethological isolation noted between the two species. I am indebted to Dr. W. F. Blair for his assistance and encouragement during this research project. The study was supported by NSF Graduate Fellowships 20200 and 31116.