Rodents produce highly variable ultrasound whistles as communication signals unlike many other mammals, who employ flow-induced vocal fold oscillations to produce sound. The role of larynx muscles in controlling sound features across different call types in ultrasound vocalization (USV) was investigated using laryngeal muscle electromyographic (EMG) activity, subglottal pressure measurements and vocal sound output in awake and spontaneously behaving Sprague-Dawley rats. Results support the hypothesis that glottal shape determines fundamental frequency. EMG activities of thyroarytenoid and cricothyroid muscles were aligned with call duration. EMG intensity increased with fundamental frequency. Phasic activities of both muscles were aligned with fast changing fundamental frequency contours, for example in trills. Activities of the sternothyroid and sternohyoid muscles, two muscles involved in vocal production in other mammals, are not critical for the production of rat USV. To test how stereotypic laryngeal and respiratory activity are across call types and individuals, sets of ten EMG and subglottal pressure parameters were measured in six different call types from six rats. Using discriminant function analysis, on average 80% of parameter sets were correctly assigned to their respective call type. This was significantly higher than the chance level. Since fundamental frequency features of USV are tightly associated with stereotypic activity of intrinsic laryngeal muscles and muscles contributing to build-up of subglottal pressure, USV provide insight into the neurophysiological control of peripheral vocal motor patterns.
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