A novel degree of sex difference in laryngeal physiology of Xenopus muelleri: behavioral and evolutionary implications

Characterizing sex and species differences in muscle physiology can contribute to a better understanding of proximate mechanisms underlying behavioral evolution. In Xenopus, the laryngeal muscle’s ability to contract rapidly and its electromyogram potentiation allows males to produce calls that are more rapid and intensity-modulated than female calls. Prior comparative studies have shown that some species lacking typical male features of vocalizations sometimes show reduced sex differences in underlying laryngeal physiology. To further understand the evolution of sexually differentiated laryngeal muscle physiology and its role in generating behavior, we investigated sex differences in the laryngeal physiology of X. muelleri, a species in which male and female calls are similar in rapidity but different with respect to intensity modulation. We delivered ethologically relevant stimulus patterns to ex vivo X. muelleri larynges to investigate their ability to produce various call patterns, and we also delivered stimuli over a broader range of intervals to assess sex differences in muscle tension and electromyogram potentiation. We found a small but statistically significant sex difference in laryngeal electromyogram potentiation that varied depending on the number of stimuli. We also found a small interaction between sex and stimulus interval on muscle tension over an ethologically relevant range of stimulus intervals; male larynges were able to produce similar tensions to female larynges at slightly smaller (11–12 ms) inter-stimulus intervals. These findings are consistent with behavioral observations and present a previously undescribed intermediate sex difference in Xenopus laryngeal muscle physiology.

Methods
All csv files including acoustic data (Pattern_IM, Pattern_Interval, Pulse_Numbers, patternAspectrogram, and patternBspectrogram) come from audio recordings from male X. muelleri only. All other files include data compiled from both males and females. Acoustic data was obtained using Audacity (v. 2.3.0, audacityteam.org) computer software and electrophysiology data was obtained using LabChart (v. 8.0.10, ADInstruments) computer software.