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  1. Maternal hormones such as estrogens deposited into the yolk of turtle eggs follow circulating levels in adult females, and they may alter the sexual fate of developing embryos in species with temperature-dependent sex determination (TSD). In temperate regions, this deposition occurs during the spring when estrogens increase in adult females as ambient temperatures rise, drop after the first clutch, and peak again (albeit less) in the fall. Global warming alters turtle nesting phenology (inducing earlier nesting), but whether it affects circulating hormones remains unknown, hindering our understanding of all potential challenges posed by climate change and the adaptive potential (or lack thereof) of turtle populations. Here, we addressed this question in painted turtles (Chrysemys picta) by quantifying estradiol, estrone, and testosterone via mass spectrometry in the blood of wild adult females exposed to 26 °C and 21 °C in captivity between mid-August and mid-October (15 females per treatment). Results from ANOVA and pairwise comparisons revealed no differences between treatments in circulating hormones measured at days 0, 2, 7, 14, 28, and 56 of the experiment. Further research is warranted (during the spring, using additional temperatures) before concluding that females are truly buffered against the indirect risk of climate change via maternal hormone allocation. 
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  2. During meiotic prophase I, tightly regulated processes take place, from pairing and synapsis of homologous chromosomes to recombination, which are essential for the generation of genetically variable haploid gametes. These processes have canonical meiotic features conserved across different phylogenetic groups. However, the dynamics of meiotic prophase I in non-mammalian vertebrates are poorly known. Here, we compare four species from Sauropsida to understand the regulation of meiotic prophase I in reptiles: the Australian central bearded dragon ( Pogona vitticeps ), two geckos ( Paroedura picta and Coleonyx variegatus ) and the painted turtle ( Chrysemys picta ). We first performed a histological characterization of the spermatogenesis process in both the bearded dragon and the painted turtle. We then analyzed prophase I dynamics, including chromosome pairing, synapsis and the formation of double strand breaks (DSBs). We show that meiosis progression is highly conserved in reptiles with telomeres clustering forming the bouquet , which we propose promotes homologous pairing and synapsis, along with facilitating the early pairing of micro-chromosomes during prophase I (i.e., early zygotene). Moreover, we detected low levels of meiotic DSB formation in all taxa. Our results provide new insights into reptile meiosis. 
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  3. Temperature-dependent sex determination (TSD) decides the sex fate of an individual based on incubation temperature. However, other environmental factors, such as pollutants, could derail TSD sexual development. Cadmium is one such contaminant of soils and water bodies known to affect DNA methylation, an epigenetic DNA modification with a key role in sexual development of TSD vertebrate embryos. Yet, whether cadmium alters DNA methylation of genes underlying gonadal formation in turtles remains unknown. Here, we investigated the effects of cadmium on the expression of two gene regulators of TSD in the painted turtle, Chrysemys picta, incubated at male-producing and female-producing temperatures using qPCR. Results revealed that cadmium alters transcription of Dmrt1 and aromatase, overriding the normal thermal effects during embryogenesis, which could potentially disrupt the sexual development of TSD turtles. Results from a preliminary DNA methylation-sensitive PCR assay implicate changes in DNA methylation of Dmrt1 as a potential cause that requires further testing (aromatase methylation assays were precluded). 
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