Implementation of a method for sperm cryopreservation in sceloporine lizards.

Actual loss of lizard biodiversity continues, even with the implementation of conventional conservation programs. An approach including assisted reproductive techniques such as sperm cryopreservation may contribute to the management of endangered species. We developed a method for sperm cryopreservation in sceloporine lizards and compared the response among the studied species. Prior to the mating season, we obtained semen from adult males of Sceloporus aeneus (n = 21), Sceloporus grammicus (n = 20) and Sceloporus torquatus (n = 21) via pressure of the genital papilla. Volume and sperm concentration were measured before semen dilution in a Tris-egg yolk (TEY) medium to evaluate progressive motility, sperm viability, morphology, plasma membrane and acrosome integrity. Then, we cooled the remaining volumes to 5°C at a rate of 0.1°C per minute to incorporate glycerol (8% v/v) in two fractions. Immediately afterwards, we placed 40 µl of the mix on solid CO2 to form pellets and immersed them in liquid nitrogen for storage. We thawed the pellets at 29°C for 3 minutes and diluted them 1:1 (v/v) in TEY medium to assess sperm quality. We found a positive relationship between body weight and seminal volume in S. grammicus and S. torquatus and a negative correlation with sperm concentration in S. grammicus (P < 0.05). Moreover, we observed that the freezing-thawing process decreased sperm quality in the three species, mostly affecting motility and viability. However, S. torquatus and S. aeneus showed a higher sperm tolerance than S. grammicus.

Sex steroids are correlated with environmental factors and body condition during the reproductive cycle in females of the lizard Sceloporus torquatus.

Reproduction is regulated by multiple factors that influence physiology and behavior to ensure the continuity of species. However, more work is needed to examine the complex relationships between environmental factors and endocrine transducers that modulate reproductive cycles, particularly in lizards. Here, we aimed to characterize the variation in plasma sex steroid levels in different stages of the reproductive cycle in the lizard Sceloporus torquatus and assess whether sex steroid levels were related to environmental factors (temperature, photoperiod, precipitation, and relative humidity) and body condition. Plasma concentrations of estradiol (E2) and progesterone (P4) from blood samples were quantified by enzyme-linked immunosorbent assay (ELISA) and radioimmunoanalysis (RIA), respectively. Our results indicate that sex steroid concentrations were positively related to follicular development but negatively related to temperature and precipitation. E2 increased as the follicles grew, and its concentrations were highest in the preovulatory phase. P4 showed a similar pattern and persisted during pregnancy. Changes in body condition were non-significant and mainly unrelated to the reproductive stage and plasma sex steroids. Our findings indicate that sex steroids change depending on the season and reproductive stage. We observed high concentrations of E2 and P4 in the late vitellogenic and preovulatory stages, probably because of their role in promoting vitellogenesis and ovulation. Additionally, we observed that follicular development is correlated with temperature and photoperiod. To better understand the mechanisms underlying reproduction, future studies of captive populations where environmental factors can be manipulated are needed.