What is the origin of the expression "to prick one's ears up"? Does this come from an actual physical action seen in animals or from some literature reference?
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It comes from the action an animal with suitable ears makes when it hears something important and tries to hear better. The first dog here has his ears pricked up, while the second does not.
When used of animals such as dogs or cats, it is a literal expression. It becomes a metaphor when it is used of persons. The Free Dictionary says:
As Daniel has pointed out, the idiom derives from observing animals. As to the etymology of the idiom itself, it has quite a nice history.
It first appeared, it seems, in 1449. There's a nice extract from Walter Bower's Scotichronicon (which, as Hugo pointed out, is a continuation of John of Fordun's Chronica Gentis Scotorum):
Prick, as a verb, seems to have been a favourite with medieval romance writers: just see how many times Chaucer plays with it in The Canterbury Tales. But Fordun's use seems to have caught on, because it's also used in the same way 75 years later in Douglas' translation of Virgil's Aeneid:
After its entry point, it seems to have had a nice continuity in the English language:
A precedent of prick-eared can be found in the late Middle English prykeryd, c.1420.
Sherman M. Kuhn's Middle English Dictionary: