MicroCT analysis unveils the role of inflatable female genitalia and male tibial complex in the genital coupling in the spider genus Aysha (Anyphaenidae, Araneae).

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Sperm transfer in spiders is achieved by copulatory organs on the male pedipalps (i.e., copulatory bulbs), which can be simple or a complex set of sclerites and membranes. During copulation, these sclerites can be used to anchor in corresponding structures in the female genitalia by means of hydraulic pressure. In the most diverse group of Entelegynae spiders, the retrolateral tibial apophysis clade, the female role in the coupling of genitalia is considered rather passive, as conformational changes of the female genital plate (i.e., the epigyne) during copulation are scarce. Here, we reconstruct the genital mechanics of two closely related species belonging to the Aysha prospera group (Anyphaenidae) that bear a membranous, wrinkled epigyne and male pedipalps with complex tibial structures. By using microcomputed tomography dataof a cryofixed mating pair, we reveal that most of the epigyne remains greatly inflated during genital coupling, and that the male tibial structures are coupled to the epigyne by the inflation of a tibial hematodocha. We propose that a turgent female vulva is a prerequisite for the genital coupling, which could implicate a female control device, and that the structures from the male copulatory bulb have been functionally replaced by tibial structures in these species. Furthermore, we show that the conspicuous median apophysis is maintained in spite of being functionally redundant, posing a puzzling situation.