The vertebrate heart is a muscular pump that contracts in a rhythmic fashion to propel the blood through the body. During evolution, the morphologically complex four-chambered heart of birds and mammals has evolved from a single-layered tube with peristaltic contractility. The heart of Drosophila, referred to as the dorsal vessel, is a blind sac composed of myogenic cells that contract rhythmically. The fish heart is composed of a single atrial chamber connected to a single ventricular chamber. The evolutionary development of fast-contracting chambers allowed the heart to build up high blood pressures. In amphibians two atrial chambers exist, separated by a septum, connecting to a single ventricle. The division of a common atrium and ventricle into right and left-sided chambers represents an evolutionary milestone in the development of the four-chambered heart and is necessary for separation of oxygenated and deoxygenated blood. In amphibians and reptiles, pulmonary and systemic circulations are incompletely separated allowing adaptable blood flows to both circulations. In contrast, the hearts of birds and mammals, in which septa completely separate the pulmonary and systemic circulations, both circulations have similar flows, but blood pressures can be regulated separately. In this review we focus, in a morphologically integrated fashion, on the molecular interactions that govern the intricate cardiac design.