The secrets of anterior pituitary development and cell lineage determination have been revealed mostly by genetic analyses. The requirement for three homeobox genes, Lhx3, Lhx4 and Titf1, during early organogenesis was proven by gene targeting. Spontaneous mouse mutations revealed two additional homeobox genes, Pit1 and Prop1, that are critical for specialization and proliferation of subsets of the five differentiated cell types. Analysis of patients with pituitary insufficiency has demonstrated the importance of these two genes in human pituitary function. Recently, several other homeobox genes have been identified and implicated in pituitary organogenesis. Genetic manipulations of these genes will undoubtedly add to the emerging genetic hierarchy regulating the ontogeny of this major hormone-producing gland.