Acquisition of novel muscles enabled protruding and retracting mechanisms of female penis in sex-role reversed cave insects

Brazilian sex-role reversed cave insects (genus Neotrogla) have a striking structure called the gynosome (or female penis), which deeply penetrates male vagina-like genitalia during copulation to receive nutritious semen. However, the protruding and retracting mechanisms of the female penis, including their evolutionary origin, are poorly understood. By using micro-computed tomography (µCT), we compared the genital morphology and musculature between species with a gynosome and others lacking this structure. As a result, we discovered two groups of muscles related to the protrusion and retraction of gynosomes. These muscles were also observed in species with non-protrusible prepenis. This suggests that evolution of these muscles preceded the acquisition of the protruding function of the gynosome, originally having a putative stimulatory function to receive nutritious semen. This intermediate stage probably allowed for the reversal of genital functions.