In his 1872 work, The Expression of Emotions in Man and Animals, Charles Darwin explores the uses of emotions to expand on his theories of evolution. He proposes three principles core to the expression of emotion. His second principle is that of antithesis, which proposes that expressions of opposite states of mind will employ opposite body postures. This theory has largely been explored in its relevance to psychological studies in human and mammalian expression. But do these parallels in the emotive expressions hold any validity, and can this concept be employed beyond the purely biological in that of design and animation? This research has two primary goals. First this research seeks to re-evaluate Darwin’s concept of antithesis using Jakob von Uexküll’s semiotic concept of Umwelt. This provides a mode by which we can draw parallels between human and general mammalian expression to see if opposites in body posture align with communicative intent and expression. Expanding upon these foundational ideas, this research secondarily seeks to evaluate if this concept can be expanded beyond the biological and look at the presence and use of antithesis in the animation of cartoons, whose primary purpose is to express emotion, or evoke it, within an audience. Particular attention will be paid to the concepts of animation as laid out in the Principles of Movement and Expression in Animation used by Disney and explicated by Disney animators Frank Thomas and Ollie Johnston in The Illusion of Life: Disney Animation (1985). The research will conclude by examining the relevance of Darwin’s theory in a wider semiotic and cultural context before moving onto its potential implications in the design process of modern animation.