The embryonic organizer is essential to determine one or more developmental polarities during chordate early development1,2. Functionally similar organizers also occur in more ancient animals3, and even in some protozoans such as Dictyostelium, in which the tip of the multicellular mound acts as an organizer4, establishing the main developmental axis, and regulating the size of the fruiting body5. However, our understanding of how the Dictyostelium organizer arises, and functions, is limited. Here we show that monoamine oxidase A (maoA), which degrades serotonin, confers the fate of an organizer to the Dictyostelium tip. Conversely, once a tip has formed, serotonin contributes to tip dominance. It inhibits further tip formation, and thus ensures the mound retains the size specified during an earlier developmental stage. Reducing the expression of maoA through RNA interference or by adding MAO specific inhibitors suppresses tip formation. Conversely, adding human MAOA enzyme, or an antagonist or antibodies against serotonin, restores tip formation in maoA knockdowns. Overexpression of maoA or adding a serotonin antagonist to the wildtype induces multiple tips from a single mound in a dose dependent manner. Using an array of genetic and molecular techniques, we show that serotonin’s inhibition of cAMP signalling and cell-cell adhesion is the basis of its regulation of tip formation. Our study demonstrates that serotonin, recently appreciated for its developmental roles in widespread phyla6, also has a novel and ancient role in the formation and function of an organizer.