Bats expand their vocal range by recruiting different laryngeal structures for echolocation and social communication.

Echolocating bats produce very diverse vocal signals for echolocation and social communication that span an impressive frequency range of 1 to 120 kHz or 7 octaves. This tremendous vocal range is unparalleled in mammalian sound production and thought to be produced by specialized laryngeal vocal membranes on top of vocal folds. However, their function in vocal production remains untested. By filming vocal membranes in excised bat larynges (Myotis daubentonii) in vitro with ultra-high-speed video (up to 250,000 fps) and using deep learning networks to extract their motion, we provide the first direct observations that vocal membranes exhibit flow-induced self-sustained vibrations to produce 10 to 95 kHz echolocation and social communication calls in bats. The vocal membranes achieve the highest fundamental frequencies (fo's) of any mammal, but their vocal range is with 3 to 4 octaves comparable to most mammals. We evaluate the currently outstanding hypotheses for vocal membrane function and propose that most laryngeal adaptations in echolocating bats result from selection for producing high-frequency, rapid echolocation calls to catch fast-moving prey. Furthermore, we show that bats extend their lower vocal range by recruiting their ventricular folds-as in death metal growls-that vibrate at distinctly lower frequencies of 1 to 5 kHz for producing agonistic social calls. The different selection pressures for echolocation and social communication facilitated the evolution of separate laryngeal structures that together vastly expanded the vocal range in bats.