First 3-D reconstruction of copulation in Lepidoptera: interaction of genitalia in Tortrix viridana (Tortricidae).

BACKGROUND: The process of copulation in Lepidoptera is understudied and poorly understood from a functional perspective. The purpose of the present paper is to study the interaction of the male and female genitalia of Tortrix viridana Linnaeus, 1758 via three-dimensional models of pairs fixed during copulation. Other techniques (confocal laser scanning microscopy, scanning electron microscopy and histology) were used to clarify the role of the organs involved in the process. RESULTS: Three-dimensional models based on micro-CT scanned copulating pairs were generated allowing visualisation of the position of the male and female counterparts, spatial changes during copulation, and the skeleto-muscular apparatus involved in the process. The male genitalia and their musculature are simplified in comparison with other lineages of the family, but the opposite is true for the female genitalia. The attachment of the couple is achieved only through flexion of the valvae, clasping the large and sclerotised sternite 7 of the female. The anal cone and socii of the male are in contact with certain parts of the anal papillae and the sterigma of the female. The long tubular vesica is inserted into the narrow posterior part of the ductus bursae. Its eversion is achieved by an increase in haemolymph pressure. A possible mechanism of stimulation of the female via pulsations of the diverticulum of the vesica was discovered. A compressed sclerotised area of the ductus bursae putatively serves as a valve controlling the transfer of ejaculated materials. Copulation progresses through two phases: in the first the vesica and its diverticulum are inflated by haemolymph, and in the second the diverticulum is not inflated, and the vesica is occupied by viscous ejaculated material. The formation of the multilayered spermatophore was observed, and we discovered that sperm is transferred very late in the copulation process. CONCLUSIONS: Copulation process in Lepidoptera is studied for the first time with three-dimensional reconstructions of couples of Tortrix viridana, used as a model species. The internal genitalia is the scenario of multiple interactions between male and female, but the external remain static. A possible mechanism of stimulation of the female internal copulation organs is proposed.

Cranial kinesis in the miniaturised lizard Ablepharus kitaibelii (Squamata: Scincidae).

Cranial kinesis refers to intracranial movements in the vertebrate skull that do not concern the jaw joint, the middle ear or the hypobranchial skeleton. Different kinds of cranial kinesis have been reported for lizards, including mesokinesis, metakinesis, amphikinesis (simultaneous mesokinesis and metakinesis) and streptostyly. Streptostyly is considered relatively widespread within lizards, whereas mesokinesis has been documented only for geckos, varanids and anguids. The present study investigated cranial kinesis in the miniaturised scincid Ablepharus kitaibelii by integrating morphological and experimental data. Based on micro computed tomography, we provide a description of skull osteology. Cranial joints were studied with histology, which results in the first detailed description of cranial joint histology for a member of the Scincidae. Taken together, the morphological data indicate a high potential for amphikinesis and streptostyly, which was also corroborated by skull manipulations. High-speed cinematography demonstrated that mesokinesis occurs during food uptake, processing and intraoral transport cycles. Bite force measurements showed prolonged and reasonably hard biting even at large gape angles. Based on these data, we formulate a model of the amphikinetic A. kitaibelii skull mechanism, which provides an extension of Frazzetta's quadric-crank model by placing a special emphasis on metakinesis. According to this model, we hypothesise that metakinetic intracranial movements may provide a means for reducing strain in jaw adductor muscles. Presented hypotheses can be addressed and tested in future studies.

Dynamic evolutionary interplay between ontogenetic skull patterning and whole-head integration.

The arrangement and morphology of the vertebrate skull reflect functional and ecological demands, making it a highly adaptable structure. However, the fundamental developmental and macroevolutionary mechanisms leading to different vertebrate skull phenotypes remain unclear. Here we exploit the morphological diversity of squamate reptiles to assess the developmental and evolutionary patterns of skull variation and covariation in the whole head. Our geometric morphometric analysis of a complex squamate ontogenetic dataset (209 specimens, 169 embryos, 44 species), covering stages from craniofacial primordia to fully ossified bones, reveals that morphological differences between snake and lizard skulls arose gradually through changes in spatial relationships (heterotopy) followed by alterations in developmental timing or rate (heterochrony). Along with dynamic spatiotemporal changes in the integration pattern of skull bone shape and topology with surrounding brain tissues and sensory organs, we identify a relatively higher phenotypic integration of the developing snake head compared with lizards. The eye, nasal cavity and Jacobson's organ are pivotal in skull morphogenesis, highlighting the importance of sensory rearrangements in snake evolution. Furthermore, our findings demonstrate the importance of early embryonic, ontogenetic and tissue interactions in shaping craniofacial evolution and ecological diversification in squamates, with implications for the nature of cranio-cerebral relations across vertebrates.

Nomenclatural-taxonomic notes on the genus Ablepharus Fitzinger, 1824 (Squamata: Scincidae).

The two following notes concern two species of small skinks, genus Ablepharus: Lindberg's Twin-striped Skink (Ablepharus lindbergi) and the Snake-eyed Skink (Ablepharus kitaibelii). Both species are small fossorial/semi-fossorial skinks with fused eyelids and a hidden lifestyle. A. kitaibelii is distributed disjunctly in several subspecies in Central and Southeastern Europe as well as in Western Turkey, A. lindbergi is distributed in Western Afghanistan (Uetz et al. 2020).