We investigated the behavioural mechanism underlying the recently discovered latitudinal variation in the social structure of the spider Anelosimus studiosus through population censuses, behavioural interaction trials, reciprocal nest transplants to different temperature environments and breeding experiments. Nest censuses completed at replicated sites at 2° changes in latitude between south Florida (26°) and east Tennessee (36°) indicated that the dominant social structure is a solitary female nest. Multifemale nests consisting of cooperative females and their young first appeared in the populations at 30° latitude and increased in frequency of representation with further increases in latitude as did number of females within multifemale nests. Interaction trials showed that communication and physical contact underlie the two social structures. Females from solitary female nests and F1 offspring reared in the laboratory demanded space (asocial phenotype), whereas individuals of multifemale nest origin (from same and foreign nests) were attracted to one another (social phenotype). Field experiments further showed that (1) individuals that dispersed from multifemale nests dispersed shorter distances and (2) nest coalescence was observed only at the higher latitudes, coincident with the presence of multifemale nests. Habitat transplants suggested that the behaviour underlying social structure is not plastic in this system, and breeding experiments eliminated a cryptic species alternative to the presence of different social structure phenotypes. Although both asocial and social phenotypes were present at all latitudes, the frequency of the social phenotype was rare at lower latitudes.