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We compared egg size phenotypes and tested several predictions from the optimal egg size (OES) and bet-hedging theories in two North American desert-dwelling sister tortoise taxa, Gopherus agassizii and G. morafkai, that inhabit different climate spaces: relatively unpredictable and more predictable climate spaces, respectively. Observed patterns in both species differed from the predictions of OES in several ways. Mean egg size increased with maternal body size in both species. Mean egg size was inversely related to clutch order in G. agassizii, a strategy more consistent with the within-generation hypothesis arising out of bet-hedging theory or a constraint in egg investment due to resource availability, and contrary to theories of density dependence, which posit that increasing hatchling competition from later season clutches should drive selection for larger eggs. We provide empirical evidence that one species, G. agassizii, employs a bet-hedging strategy that is a combination of two different bet-hedging hypotheses. Additionally, we found some evidence for G. morafkai employing a conservative bet-hedging strategy. (e.g., lack of intra- and interclutch variation in egg size relative to body size). Our novel adaptive hypothesis suggests the possibility that natural selection favors smaller offspring in late-season clutches because they experience a more benign environment or less energetically challenging environmental conditions (i.e., winter) than early clutch progeny, that emerge under harsher and more energetically challenging environmental conditions (i.e., summer). We also discuss alternative hypotheses of sexually antagonistic selection, which arise from the trade-offs of son versus daughter production that might have different optima depending on clutch order and variation in temperature-dependent sex determination (TSD) among clutches. Resolution of these hypotheses will require long-term data on fitness of sons versus daughters as a function of incubation environment, data as yet unavailable for any species with TSD. [ABSTRACT FROM AUTHOR]

Sex-specific maternal effects can be adaptive sources of phenotypic plasticity. Reptiles with temperature-dependent sex determination ( TSD) are a powerful system to investigate such maternal effects because offspring phenotype, including sex, can be sensitive to maternal influences such as oestrogens and incubation temperatures., In red-eared slider turtles ( Trachemys scripta), concentrations of maternally derived oestrogens and incubation temperatures increase across the nesting season; we wanted to determine if sex ratios shift in a seasonally concordant manner, creating the potential for sex-specific maternal effects, and to define the sex ratio reaction norms under fluctuating temperatures across the nesting season., Eggs from early and late season clutches were incubated under a range of thermally fluctuating temperatures, maternally derived oestradiol concentrations were quantified via radioimmunoassay, and hatchling sex was identified. We found that late-season eggs had higher maternal oestrogen concentrations and were more likely to produce female hatchlings. The sex ratio reaction norm curves systematically varied with season, such that with even a slight increase in temperature (0·5 °C), late-season eggs produced up to 49% more females than early-season eggs., We found a seasonal shift in sex ratios which creates the potential for sex-specific phenotypic matches across the nesting season driven by maternal effects. We also describe, for the first time, systematic variation in the sex ratio reaction norm curve within a single population in a species with TSD., A is available for this article. [ABSTRACT FROM AUTHOR]

Higher temperatures associated with climate change have the potential to significantly alter the population sex ratio of species with temperature-dependent sex determination. Whether or not elevated temperature affects sex determination depends on both the absolute temperature experienced and the stage of development at which the thermal conditions occur. We explored the importance of exposure timing during early development in the coral reef fish, Acanthochromis polyacanthus, by increasing water temperature 1.5 or 3 °C above the summer average (28.5 °C) at different stages of development. We also measured the effect of treatment temperature on fish size and condition, in order to gauge how the thermal threshold for sex-ratio bias may compare with other commonly considered physiological metrics. Increasing grow-out temperature from 28.5 to 30 °C had no effect on the sex ratio of offspring, whereas an increase to 31.5 °C (+3 °C) produced a strong male bias (average ~90%). The thermosensitive period for this species lasted between 25 and 60 d post hatching, with the bias in sex ratio greater the earlier that fish experienced warm conditions. Temperatures high enough to bias the sex ratio are likely to be seen first during late summer (January and February) and would affect clutches produced late in the breeding season. There was no change to fish condition in response to temperature; however, the two higher temperature treatments produced significantly smaller fish at sampling. Clutches produced early in the season could buffer the population from a skewed sex ratio, as their development will remain below the thermal threshold; however, continued ocean warming could mean that clutches produced earlier in the breeding season would also be affected in the longer term. A skewed sex ratio could be detrimental to population viability by reducing the number of females in the breeding population. [ABSTRACT FROM AUTHOR]

In many reptile species, gonadal sex is affected by environmental temperature during a critical period of embryonic development—a process known as temperature-dependent sex determination (TSD). 1 The oviparous red-eared slider turtle, Trachemys scripta , has a warm-female/cool-male TSD system and is among the best-studied members of this group. 2 When incubated at low temperatures, the somatic cells of the bipotential gonad differentiate into Sertoli cells, the support cells of the testis, whereas at high temperatures, they differentiate into granulosa cells, the support cells of the ovary. 3 Here, we report the unexpected finding that temperature independently affects the number of primordial germ cells (GCs) in the embryonic gonad at a time before somatic cell differentiation has initiated. Specifically, embryos incubated at higher, female-inducing temperatures have more GCs than those incubated at the male-inducing temperature. Furthermore, elimination of GCs in embryos incubating at intermediate temperatures results in a strong shift toward male-biased sex ratios. This is the first evidence that temperature affects GC number and the first evidence that GC number influences sex determination in amniotes. This observation has two important implications. First, it supports a new model in which temperature can impact sex determination in incremental ways through multiple cell types. Second, the findings have important implications for a major unresolved question in the fields of ecology and evolutionary biology—the adaptive significance of TSD. We suggest that linking high GC number with female development improves female reproductive potential and provides an adaptive advantage for TSD. • Higher (female) incubation temperature directly increases germ cell (GC) number • Depletion of GCs at intermediate temperature leads to male-biased sex ratios • Findings support a new TSD model in which germ cells influence sex determination • Coupling GC number with female development improves reproductive fitness Tezak et al. find that warmer incubation temperatures directly increase germ cell number in the red-eared slider turtle and provide the first evidence that germ cells can influence sex determination in amniotes. This observation supports a new model in which temperature can impact sex determination in incremental ways through multiple cell types. [ABSTRACT FROM AUTHOR]

The Chinese alligator Alligator sinensis is one of the most endangered crocodilian species, and typically exhibits temperature-dependent sex determination. It is extremely important to clarify the sex structure of Chinese alligators to implement recovery projects successfully. However, the sex ratio of wild Chinese alligators remains unknown. In this study, we collected 28 years of sex ratio data from Chinese alligators residing in the natural and artificial habitats of Changxing Nature Reserve, China, and examined the differences in the sex ratio dynamics between these two populations.We observed that the sex ratio of wild Chinese alligators is 1 male to 4.507 females, which was significantly lower compared to that of the captive population (1 to 2.040; P < 0.001), and is significantly different to previously documented sex ratios for this species (all P < 0.01). Furthermore, we documented an annually stable (P = 1.000) female-biased sex ratio for wild alligators at hatching [1 male to 4.747 females; 0.174 (0.167–0.182)], in contrast to a dramatically fluctuating sex ratio (P < 0.001) in captivity [1 male to 1.674 females; 0.374 (0.246–0.593)], showing a potential mechanism for adjusting the sex structure. Finally, we found that the hatchling sex ratios were similar to that of the population sex ratio (P = 0.748), with little correlation to air temperature values in the 60–70 day incubation period during the breeding season (July and August; both P > 0.05). Overall, this study indicates that the stabilized female-biased sex ratio of Changxing Chinese alligators might result from selection pressure caused by local mate competition and major inbreeding [Current Zoology 59 (6) : 725–731, 2013 ].

AbstractSex chromosomes rapidly turn over in several taxonomic groups. Sex chromosome turnover is generally thought to start with the appearance of a new sex-determining gene on an autosome while an old sex-determining gene still exists, followed by the fixation of the new one. However, we do not know how prevalent the transient state is, where multiple sex-determining loci coexist within natural populations. Here, we removed a Y chromosome with a master male-determining gene DMY from medaka fish using high temperature-induced sex-reversed males. After four generations, the genomic characteristics of a sex chromosome were found on one chromosome, which was an autosome in the original population. Thus, the elimination of a master sex-determining locus can reveal a cryptic locus with a possible sex-determining effect, which can be the seed for sex chromosome turnover. Our results suggest that populations that seem to have a single-locus XY system may have other chromosomal regions with sex-determining effects. In conclusion, the coexistence of multiple sex-determining genes in a natural population may be more prevalent than previously thought. Experimental elimination of a master sex-determining locus may serve as a promising method for finding a locus that can be a protosex chromosome.

Over the past two decades, there has been an astounding growth in the documentation of vertebrate facultative parthenogenesis (FP). This unusual reproductive mode has been documented in birds, non-avian reptiles-specifically lizards and snakes-and elasmobranch fishes. Part of this growth among vertebrate taxa is attributable to awareness of the phenomenon itself and advances in molecular genetics/genomics and bioinformatics, and as such our understanding has developed considerably. Nonetheless, questions remain as to its occurrence outside of these vertebrate lineages, most notably in Chelonia (turtles) and Crocodylia (crocodiles, alligators and gharials). The latter group is particularly interesting because unlike all previously documented cases of FP in vertebrates, crocodilians lack sex chromosomes and sex determination is controlled by temperature. Here, using whole-genome sequencing data, we provide, to our knowledge, the first evidence of FP in a crocodilian, the American crocodile, Crocodylus acutus . The data support terminal fusion automixis as the reproductive mechanism; a finding which suggests a common evolutionary origin of FP across reptiles, crocodilians and birds. With FP now documented in the two main branches of extant archosaurs, this discovery offers tantalizing insights into the possible reproductive capabilities of the extinct archosaurian relatives of crocodilians and birds, notably members of Pterosauria and Dinosauria.

For species with temperature-dependent sex determination (TSD) there is the fear that rising temperatures may lead to single-sex populations and population extinction. We show that for sea turtles, a major group exhibiting TSD, these concerns are currently unfounded but may become important under extreme climate warming scenarios. We show how highly female-biased sex ratios in developing eggs translate into much more balanced operational sex ratios so that adult male numbers in populations around the world are unlikely to be limiting. Rather than reducing population viability, female-biased offspring sex ratios may, to some extent, help population growth by increasing the number of breeding females and hence egg production. For rookeries across the world (n = 75 sites for seven species), we show that extreme female-biased hatchling sex ratios do not compromise population size and are the norm, with a tendency for populations to maximize the number of female hatchlings. Only at extremely high incubation temperature does high mortality within developing clutches threaten sea turtles. Our work shows how TSD itself is a robust strategy up to a point, but eventually high mortality and female-only hatchling production will cause extinction if incubation conditions warm considerably in the future. [ABSTRACT FROM AUTHOR]

Temperature dependent sex determination (TSD) is the process by which the environmental temperature experienced during embryogenesis influences the sex of an organism, as in the red-eared slider turtle Trachemys scripta elegans . In accord with current paradigms of vertebrate sex determination, temperature is believed to exert its effects on sexual development in T. scripta entirely within the middle third of development, when the gonad is forming. However, whether temperature regulates the transcriptome in T. scripta early embryos in a manner that could influence secondary sex characteristics or establish a pro-male or pro-female environment has not been investigated. In addition, apart from a handful of candidate genes, very little is known about potential similarities between the expression cascade during TSD and the genetic cascade that drives mammalian sex determination. Here, we conducted an unbiased transcriptome-wide analysis of the effects of male- and female-promoting temperatures on the turtle embryo prior to gonad formation, and on the gonad during the temperature sensitive period. We found sexually dimorphic expression reflecting differences in steroidogenic enzymes and brain development prior to gonad formation. Within the gonad, we mapped a cascade of differential expression similar to the genetic cascade established in mammals. Using a Hidden Markov Model based clustering approach, we identified groups of genes that show heterochronic shifts between M. musculus and T. scripta . We propose a model in which multiple factors influenced by temperature accumulate during early gonadogenesis, and converge on the antagonistic regulation of aromatase to canalize sex determination near the end of the temperature sensitive window of development. [ABSTRACT FROM AUTHOR]

The environment surrounding the embryos has a profound impact on the developmental process and phenotypic outcomes of the organism. In species with temperature-dependent sex determination, gonadal sex is determined by the incubation temperature of the eggs. A mechanistic link between temperature and transcriptional regulation of developmental genes, however, remains elusive. In this study, we examine the changes in DNA methylation and histone modification patterns of the aromatase (cyp19a1) gene in embryonic gonads of red-eared slider turtles (Trachemys scripta) subjected to a temperature shift during development. Shifting embryos from a male-producing temperature (MPT) to a female-producing temperature (FPT) at the beginning of the temperature-sensitive period (TSP) resulted in an increase in aromatase mRNA expression while a shift from FPT to MPT resulted in decreased expression. DNA methylation levels at CpG sites in the promoter of the aromatase gene were high (70–90%) at the beginning of TSP, but decreased in embryos that were incubated at constant FPT and those shifted from MPT to the FPT. This decrease in methylation in the promoter inversely correlated with the expected increase in aromatase expression at the FPT. The active demethylation under the FPT was especially prominent at the CpG site upstream of the gonad-specific TATA box at the beginning of TSP and spread downstream of the gene including exon1 as the gonad development progressed. In embryos incubated at FPT, the promoter region was also labeled by canonical transcriptional activation markers, H3K4me3 and RNA polymerase II. A transcriptional repression marker, H3K27me3, was observed in temperature-shifted gonads of both temperature groups, but was not maintained throughout the development in either group. Our findings suggest that DNA hypomethylation and H3K4me3 modification at the aromatase promoter may be a primary mechanism that releases a transcriptional block of aromatase to initiate a cascade of ovarian differentiation. [ABSTRACT FROM AUTHOR]

Dr. Louis J. Guillette Jr. thought of himself as a reproductive biologist. However, his interest in reproductive biology transcended organ systems, life history stages, species, and environmental contexts. His integrative and collaborative nature led to diverse and fascinating research projects conducted all over the world. He doesn’t leave us with a single legacy. Instead, he entrusts us with several. The purpose of this review is to highlight those legacies, in both breadth and diversity, and to illustrate Dr. Guillette’s grand contributions to the field of reproductive biology. He has challenged the field to reconsider how we think about our data, championed development of novel and innovative techniques to measure endocrine function, helped define the field of endocrine disruption, and lead projects to characterize new endocrine disrupting chemicals. He significantly influenced our understanding of evolution, and took bold and important steps to translate all that he has learned into advances in human reproductive health. We hope that after reading this manuscript our audience will appreciate and continue Dr. Guillette’s practice of open-minded and passionate collaboration to understand the basic mechanisms driving reproductive physiology and to ultimately apply those findings to protect and improve wildlife and human health. [ABSTRACT FROM AUTHOR]

We hypothesize that under the predicted scenario of climate change epigenetically mediated environmental sex determination could become an epigenetic trap. Epigenetically regulated environmental sex determination is a mechanism by which species can modulate their breeding strategies to accommodate environmental change. Growing evidence suggests that epigenetic mechanisms may play a key role in phenotypic plasticity and in the rapid adaptation of species to environmental change, through the capacity of organisms to maintain a non-genetic plastic memory of the environmental and ecological conditions experienced by their parents. However, inherited epigenetic variation could also be maladaptive, becoming an epigenetic trap. This is because environmental sex determination can alter sex ratios by increasing the survival of one of the sexes at the expense of negative fitness consequences for the other, which could lead not only to the collapse of natural populations, but also have an impact in farmed animal and plant species. [ABSTRACT FROM AUTHOR]

Sex determination in vertebrates depends on the expression of a conserved network of genes. Sea turtles such as Lepidochelys olivacea have temperature-dependent sex determination. The present work analyses some of the epigenetic processes involved in this. We describe sexual dimorphism in global DNA methylation patterns between ovaries and testes of L. olivacea and show that the differences may arise from a combination of DNA methylation and demethylation events that occur during sex determination. Irrespective of incubation temperature, 5-hydroxymethylcytosine was abundant in the bipotential gonad; however, following sex determination, this modification was no longer found in pre-Sertoli cells in the testes. These changes correlate with the establishment of the sexually dimorphic DNA methylation patterns, down regulation of Sox9 gene expression in ovaries and irreversible gonadal commitment towards a male or female differentiation pathway. Thus, DNA methylation changes may be necessary for the stabilization of the gene expression networks that drive the differentiation of the bipotential gonad to form either an ovary or a testis in L. olivacea and probably among other species that manifest temperature-dependent sex determination. [ABSTRACT FROM AUTHOR]

Effects of xenobiotics can be organizational, permanently affecting anatomy during embryonic development, and/or activational, influencing transitory actions during adulthood. The organizational influence of endocrine-disrupting contaminants (EDC’s) produces a wide variety of reproductive abnormalities among vertebrates that exhibit temperature-dependent sex determination (TSD). Typically, such influences result in subsequent activational malfunction, some of which are beneficial in aquaculture. For example, 17-αmethyltestosterone (MT), a synthetic androgen, is utilized in tilapia farming to bias sex ratio towards males because they are more profitable. A heavily male-biased hatchling sex ratio is reported from a crocodile population near one such tilapia operation in Guanacaste, Costa Rica. In this study we test the effects of MT on sexual differentiation in American alligators, which we used as a surrogate for all crocodilians. Experimentally, alligators were exposed to MT in ovo at standard ecotoxicological concentrations. Sexual differentiation was determined by examination of primary and secondary sex organs post hatching. We find that MT is capable of producing male embryos at temperatures known to produce females and demonstrate a dose-dependent gradient of masculinization. Embryonic exposure to MT results in hermaphroditic primary sex organs, delayed renal development and masculinization of the clitero-penis (CTP). [ABSTRACT FROM AUTHOR]

Alterations have occurred and continue to manifest in the Earth’s biota as a result of climate change. Animals exhibiting temperature dependent sex determination (TSD), including sea turtles, are perhaps most vulnerable to a warming of the Earth as highly skewed sex ratios can result, potentially leading to population extinction resulting from decreased male recruitment. Recent studies have begun to quantify climate change impacts to sea turtle populations, especially in terms of predicting effects on hatchling sex ratios. However, given the inherent difficulty in studying sex ratios at this life stage, a more accurate assessment of changes in population sex ratios might be derived by evaluating the juvenile portion of foraging aggregations. We investigated the long-term trend in sex ratio of a juvenile loggerhead (Caretta caretta) sea turtle population inhabiting Pamlico and Core Sounds, North Carolina, USA. We used plasma testosterone reference ranges measured using radioimmunoassay (RIA) to assign sex for 959 turtles and confirmed sex assignment of a subset (N = 58) of the sampled turtles through laparoscopic examination of their gonads. Our results demonstrate that for this particular population of loggerheads, sex ratios (3Females:1Male) had not significantly changed over a 10 year period (1998–2007), nor showed any significant difference among 5-cm straight carapace length (SCL) size classes. Ultimately, these findings provide a basis for comparison with future sex ratios, and highlight the importance of establishing similar long-term studies monitoring secondary, rather than primary, sex ratios, so that needed mitigation measures to climate change impacts can be implemented. [ABSTRACT FROM AUTHOR]

Spatial variation in global climate change makes population-specific responses to this enigmatic threat pertinent on a regional scale. Organisms with temperature-dependent sex determination (TSD) potentially possess a unique physiological susceptibility that threatens population viability if rapid environmental effects on sex ratios render populations non-viable. A heavily male-biased sex ratio for hatchling American crocodiles of the Tempisque Basin, Costa Rica requires assessment of how nest temperature affects sex determination at this site, how females might compensate for these effects when creating nests, and how current patterns of climate change might alter future sex ratios and survival in hatchling cohorts. We demonstrate high within-nest variation in temperature but predict a female bias at hatching based on nest temperatures quantified here. Further, our data suggest that egg size and metabolic heating associated with this factor outweighs microhabitat parameters and depth in influencing nest thermal regimes. Finally, we document regional warming in the Tempisque Basin over the last 15 years and project that further heating over the next 15 years will not yield hatchling sex ratios as male biased as those currently found at this site. Thus, we find no support for nest temperature or climate change as likely explanations for male-biased American crocodile ( Crocodylus acutus ) sex ratios in the Tempisque Basin. [ABSTRACT FROM AUTHOR]

The environment experienced by parents can impact the phenotype of their offspring (parental effects), a critical component of organismal ecology and evolution in variable or changing environments. Although temperature is a central feature of the environment for ectotherms, its role in parental effects has been little explored until recently. Here, parental basking opportunity was manipulated in an oviparous lizard with temperature-dependent sex determination, the jacky dragon (Amphibolurus muricatus). Eggs were incubated at a temperature that typically produces a 50:50 sex ratio, and hatchlings were reared in a standard thermal environment. Offspring of parents in short bask conditions appeared to have better fitness outcomes in captive conditions than those of parents in long bask conditions -- they had greater growth and survival as a function of their mass. In addition, the sex of offspring (male or female) depended on the interaction between parental treatment and egg mass, and treatment impacted whether sons or daughters grew larger in their first season. The interactive effects of treatment on offspring sex and growth are consistent with adaptive explanations for the existence of temperature-dependent sex determination in this species. Moreover, the greater performance recorded in short bask offspring may represent an anticipatory parental effect to aid offspring in predicted conditions of restricted thermal opportunity. Together, these responses constitute a crucial component of the population response to spatial or temporal variation in temperature. [ABSTRACT FROM AUTHOR]

Vertebrate sex-determining mechanisms ( SDMs) are triggered by the genotype ( GSD), by temperature ( TSD), or occasionally, by both. The causes and consequences of SDM diversity remain enigmatic. Theory predicts SDM effects on species diversification, and life-span effects on SDM evolutionary turnover. Yet, evidence is conflicting in clades with labile SDMs, such as reptiles. Here, we investigate whether SDM is associated with diversification in turtles and lizards, and whether alterative factors, such as lifespan's effect on transition rates, could explain the relative prevalence of SDMs in turtles and lizards (including and excluding snakes). We assembled a comprehensive dataset of SDM states for squamates and turtles and leveraged large phylogenies for these two groups. We found no evidence that SDMs affect turtle, squamate, or lizard diversification. However, SDM transition rates differ between groups. In lizards TSD-to- GSD surpass GSD-to- TSD transitions, explaining the predominance of GSD lizards in nature. SDM transitions are fewer in turtles and the rates are similar to each other ( TSD-to- GSD equals GSD-to- TSD), which, coupled with TSD ancestry, could explain TSD's predominance in turtles. These contrasting patterns can be explained by differences in life history. Namely, our data support the notion that in general, shorter lizard lifespan renders TSD detrimental favoring GSD evolution in squamates, whereas turtle longevity permits TSD retention. Thus, based on the macro-evolutionary evidence we uncovered, we hypothesize that turtles and lizards followed different evolutionary trajectories with respect to SDM, likely mediated by differences in lifespan. Combined, our findings revealed a complex evolutionary interplay between SDMs and life histories that warrants further research that should make use of expanded datasets on unexamined taxa to enable more conclusive analyses. [ABSTRACT FROM AUTHOR]

Species with temperature-dependent sex determination (TSD) are more vulnerable to climate change than species with genetically based sex determination because an increase in average environmental temperature could lead to biased offspring sex ratios. In TSD species, the relationship between sex ratio and constant incubation temperature is characterized by two parameters: the pivotal temperature (Tpiv) and the transitional range of temperature (TRT) that produces both sexes. Species with variation in either of these parameters should have more potential to adapt to new climatic conditions, therefore putting them at lower risk to global warming. In this study, we incubated eggs of the turtle Podocnemis lewyana (Testudines: Podocnemididae) from two geographically isolated populations (Magdalena and Sinú rivers) under controlled microclimatic conditions to corroborate the TSD pattern previously reported for this species and to estimate the degree of inter- and intrapopulation variation in Tpiv and TRT. We used these estimates, together with information found in the literature on other life history traits of P. lewyana, to evaluate its adaptive potential or vulnerability to climate change. We found interpopulation variation in TRT and intrapopulation variation in Tpiv. This latter parameter exhibited a high broad-sense heritability, demonstrating this species' potential to adapt to climate change; however, when considering the narrow TRT found in this study, the species' equatorial distribution and related adaptations to relatively more stable climates, its fragmented populations and low vagility that prevent gene flow, and its low genetic variability, we concluded that the ability of P. lewyana to adapt to climate change is limited. Las especies con determinación sexual dependiente de la temperatura (DST) son más vulnerables al cambio climático que aquellas con determinación sexual genética, ya que un aumento en la temperatura ambiental puede llevar a sesgos en las proporciones sexuales de la progenie. En especies con DST la relación entre las proporciones sexuales y la temperatura constante de incubación, está caracterizada por dos parámetros: la temperatura pivotal (Tpiv) y el rango de transición de temperaturas (RTT) en el que se producen ambos sexos. Las especies con más variación en alguno de estos parámetros tienen más potencial para adaptarse a nuevas condiciones climáticas, y en consecuencia ser menos vulnerables al calentamiento global. En este estudio incubamos huevos de la tortuga Podocnemis lewyana provenientes de dos poblaciones geográficamente aisladas (ríos Sinú y Magdalena) bajo condiciones microclimáticas controladas para corroborar la presencia de DST anteriormente registrada para esta especie, y para estimar el grado de variación en la Tpiv y el RTT entre y dentro de las poblaciones. Usamos esos estimaciones con información publicada sobre otros rasgos de historia de vida de P. lewyana para evaluar su potencial adaptivo o vulneribilidad al cambio climático. Encontramos variación interpoblacional en el RTT y variación intrapoblacional en la Tpiv. Este último parámetro exhibió un alto nivel de heredabilidad de sentido amplio demostrando el potencial que esta especie tiene para adaptarse al calentamiento global. Sin embargo considerando el RTT tan estrecho encontrado, su distribución tropical y en consecuencia adaptación a climas estables, la fragmentación de sus poblaciones y baja movilidad que previenen el flujo genético y su baja variabilidad genética, concluimos que la habilidad de P. lewyana para adaptarse al cambio climático es limitada. [ABSTRACT FROM AUTHOR]

In species with temperature-dependent sex determination, the embryo commits to its sexual fate during a time window called the thermosensitive period (TSP). Although substantial research has focused on the effect of the temperature during this canonical TSP, the effect of temperatures experienced before this time (pre-TSP) on the onset and duration of the TSP is less understood. Here, we incubated eggs of the endangered Magdalena River turtle, Podocnemis lewyana, under 3 relatively constant temperatures and 6 shift-twice treatments. Constant treatments included two 100% masculinizing temperatures that fostered a relatively slower and faster embryo development and one 100% feminizing temperature. The shift treatments included a masculinizing temperature and later a feminizing temperature for a period of time at different incubation stages. Sex ratios were determined by a geometric-morphometric sexing approach developed for this species and validated by gonadal histology. This technique is a nonlethal sexing alternative; thus, it contributes to the conservation of this endangered species. The sex ratios obtained under constant temperatures were more feminized than the sex ratio of another population native to a warmer climate. Shift-twice experiments revealed that pre-TSP temperatures affected the duration of the TSP and as such influenced sex determination. Specifically, at 29°C pre-TSP, the TSP lasted 20 d (28% of the incubation period), whereas at 31°C pre-TSP, the TSP was extended for 30 d (52% of the incubation period). An approximation of developmental stages based on the congeneric Podocnemis expansa is provided. Conservation programs should monitor temperatures throughout the entire incubation period; otherwise important effects on sex ratios and other traits may be obscured. En especies con determinación sexual termo-dependiente el embrión define su sexo durante una ventana de tiempo llamada período termosensible (PTS). Muchas investigaciones se han enfocado en el efecto de la temperatura durante este PTS canónico, pero el efecto de temperaturas ocurridas en estadíos más tempranos (pre-PTS) sobre el inicio y duración del PTS ha sido poco estudiado. Nosotros incubamos huevos de la Tortuga del Río Magdalena, Podocnemis lewyana, bajo tres temperaturas relativamente constantes y seis tratamientos de cambio térmico doble. Los tratamientos de temperaturas constantes incluyeron dos temperaturas 100% masculinizantes que inducen un desarrollo embrionario lento y rápido respectivamente, y una temperatura 100% feminizante. Los tratamientos de doble cambio incluyeron dichas temperaturas masculinizantes (pre-PTS) y un pulso de temperatura feminizante en diferentes momentos del período de incubación. Las proporciones sexuales fueron calculadas mediante una técnica de morfometría geométrica desarrollada para esta especie y validadas usando histología gonadal. Esta técnica es una alternativa de sexaje no letal que contribuye a la conservación de esta especie amenazada. Las proporciones sexuales obtenidas a temperaturas constantes fueron más feminizadas que las reportadas en otra población que habita en un clima más cálido. En los tratamientos de doble cambio las temperaturas pre-PTS afectaron la duración del PTS, y por tanto la determinación sexual. Específicamente, a 29°C pre-PTS el PTS duró 20 días (28% del periodo de incubación), mientras que a 31°C pre-PTS, el PTS se extendió a 30 días (52% del período de incubación). Los estadíos embrionarios alcanzados durante la incubación fueron estimados usando como modelo la especie congenérica, Podocnemis expansa. Sugerimos que los programas de conservación monitoreen las temperaturas durante todo el período de incubación para evitar descartar importantes efectos sobre el sexo y otros fenotipos inducidos por las temperaturas tempranas. Palabras Clave: Colombia, conservación de reptiles en peligro de extinción, determinación sexual, periodo termosensible, temperatura de incubación, tortuga del Río Magdalena, sexaje por morfometría geométrica [ABSTRACT FROM AUTHOR]

Although the adaptive significance of temperature-dependent sex determination (TSD) remains a puzzle, recent models implicate a seasonal bias in offspring sex production that translates into sex-specific fitness benefits later in life. Sex-specific emergence has been linked to fitness gains in some fish, birds and reptiles, but field data supporting the occurrence of a seasonal pattern of sex ratios in oviparous lizards are lacking. We tested the hypothesis that patterns of nest site selection and seasonal temperature changes combine to inhibit the materialization of sex-biased hatching times in a population of water dragons ( Intellagama lesueurii). As predicted, a seasonal increase in air and nest temperatures resulted in a sex bias by nesting date; male-producing clutches were laid 17.8 days sooner than female-producing clutches, on average. However, the seasonal ramping of nest temperatures also caused shorter relative incubation periods in the later, all-female clutches. As a consequence of this developmental 'catch-up', the mean hatching date for male-producing nests preceded the mean hatching date for female-producing nests by only 7.2 days. We suggest that a contracted distribution of hatching dates compared to the distribution of oviposition dates represents a general pattern for oviparous reptiles in seasonal climates, which in TSD species may largely offset the temporal disparity in nesting dates between the sexes. Although data are needed for other TSD species, such minor age differences between male and female hatchlings may not translate into fitness differences later in life, an assumption of some models for the evolution and maintenance of TSD. [ABSTRACT FROM AUTHOR]

Average air temperature is predicted to rise by at least 3 °C across the 21st century. As individual sex (male or female) is determined by temperature in many reptiles, there are concerns that climate warming will skew offspring sex ratios and local species extinctions will follow. Range shift away from hotter areas through dispersal may prevent species extinctions in many reptiles with temperature-dependent sex determination (TSD), and could be facilitated or impeded by sex-biased populations at the expanding edge. We used a simulation model to examine the role of sex-determining mechanism [TSD and genotypic sex determination (GSD)], climate warming and dispersal in determining range shift and population growth in reptiles. Dispersal influenced range shift (after climate warming) in TSD species to a greater extent than in GSD species. Our novel finding is that biased sex ratios may influence range shift, through the mixing of the rare sex (females) with males located at the colder range edges, as both sexes disperse. However, if faced with climate warming of 3 °C over the next 100 years many TSD reptiles will show limited capacity for range shift. [ABSTRACT FROM AUTHOR]

Temperature-dependent sex determination (TSD) was described nearly 50 years ago. Researchers have since identified many genes that display differential expression at male- vs. female-producing temperatures. Yet, it is unclear whether these genes (1) are involved in sex determination per se, (2) are downstream effectors involved in differentiation of ovaries and testes, or (3) are thermo-sensitive but unrelated to gonad development. Here we present multiple lines of evidence linking to sex determination in CIRBP the snapping turtle, Chelydra serpentina. We demonstrate significant associations between a single nucleotide polymorphism (SNP) (c63A > C) in CIRBP, transcript levels in embryonic gonads during specification of gonad fate, and sex in hatchlings from a thermal regime that produces mixed sex ratios. The A allele was induced in embryos exposed to a female-producing temperature, while expression of the C allele did not differ between female- and male-producing temperatures. In accord with this pattern of temperature-dependent, allele-specific expression, AA homozygotes were more likely to develop ovaries than AC heterozygotes, which, in turn, were more likely to develop ovaries than CC homozygotes. Multiple regression using SNPs in CIRBP and adjacent loci suggests that c63A > C may be the causal variant or closely linked to it. Differences in CIRBP allele frequencies among turtles from northern Minnesota, southern Minnesota, and Texas reflect small and large-scale latitudinal differences in TSD pattern. Finally, analysis of CIRBP protein localization reveals that CIRBP is in a position to mediate temperature effects on the developing gonads. Together, these studies strongly suggest that CIRBP is involved in determining the fate of the bipotential gonad. [ABSTRACT FROM AUTHOR]

Understanding the interplay between thermal variations and sex ratio in reptiles with temperature-dependent sex determination is the first step for developing long-term conservation strategies. In case of crocodilians, the information is fragmentary and insufficient for establishing a general framework to consider how thermal fluctuation influence sex determination under natural conditions. The main goal of this study was to analyze thermal variation in nests of Crocodylus moreletii and to discuss the potential implications for predicting offspring sex ratio. The study was carried out at the Centro de Estudios Tecnológicos del Mar N° 2 and at the Sistemas Productivos Cocodrilo, Campeche, Mexico. Data was collected in the nesting season of Morelet’s Crocodiles during three consecutive seasons (2007-2009). Thermal fluctuations for multiple areas of the nest chamber were registered by data loggers. We calculate the constant temperature equivalent based on thermal profiles among nests to assess whether there are differences between the nest temperature and its equivalent to constant temperature. We observed that mean nest temperature was only different among nests, while daily thermal fluctuations vary depending on the depth position within the nest chamber, years and nests. The constant temperature equivalent was different among and within nests, but not among survey years. We observed differences between constant temperature equivalent and mean nest temperature both at the top and in the middle of the nest cavities, but were not significantly different at the bottom of nest cavities. Our results enable examine and discuss the relevance of daily thermal fluctuations to predict sex ratio of the Morelet’s Crocodile. [ABSTRACT FROM AUTHOR]

The gonadal description of the freshwater atherinopsid pike silverside Chirostoma estor suggests that the gonads differentiate as ovaries or testes by 8 weeks after hatching when raised at 21 °C. Thermal treatments at 14 °C, 21 °C and 29 °C applied from fertilisation, clearly affected phenotypic sex ratios, suggesting that the thermolabile window of sex determination occurred early in development. In this study, exposure to the highest temperature led to male-biased sex ratios in this species. However, the effects of the lower and medium temperatures on the sex ratios were less clear, suggesting the presence of a mixture of genotypic and temperature-dependent sex determination (TSD) mechanisms in C. estor, similar to other atherinopsids. This work further enhances our knowledge regarding the diversity and plasticity of TSD mechanisms in atherinopsid and teleost fish. [ABSTRACT FROM AUTHOR]

The article reports that the Species threatened with extinction due to climate change need genetic diversity in order to adapt, but genetic diversity is lost as populations decline and fragment.

Climate change threatens endemic island ectothermic reptiles that display small population sizes and temperature-dependent sex determination (TSD). Studies of captive Galapagos tortoises demonstrate type A TSD with warmer incubation temperatures producing females. However, there are few published data from free-living Galapagos tortoises on incubation temperature regimes, and none on hatchling sex ratios in the wild or the potential impacts of climate change on future sex ratios. We sought to address these deficits by quantifying incubation temperatures of nests and sex ratios of juvenile tortoises along an elevation gradient on Santa Cruz Island. We focused on three geographically separated nesting zones with mean elevations of 14 m (lower), 57 m (middle), and 107 m (upper) above sea level. Nest temperatures in 54 nests distributed across the three nesting zones were measured every 4 h throughout the incubation period using iButton thermochrons. We used coelioscopy to conduct visual exams of gonads to determine the sex of 40 juvenile tortoises from the three nesting zones. During the middle trimester of incubation, the period during which sex is determined in turtles, mean nest temperatures were 25.75°C (SD = 1.08) in the upper zone, and 27.02°C (SD = 1.09), and 27.09°C (SD = 0.85) in the middle and lower zones, respectively. The proportion of juveniles that was male increased from 11.1% in the lower zone and 9.5% in the middle zone, to 80% in the upper zone. A ca. 50 m increase in elevation induced a decrease of >1.25°C in mean nest temperature during the second trimester of incubation. Over the same elevation change, the proportion of males in the juvenile tortoise population increased by ca. 70%. Temperatures on Galapagos are predicted to increase by 1-4°C over the next 50 years, which is likely to increase the frequency of female tortoises across the archipelago., Competing Interests: We declare that we have no competing interests., (© 2023 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

Genetic sex determination (SD) in most vertebrates is controlled by a single master sex gene, which ensures a 1:1 sex ratio. However, more complex systems abound, and several have been ascribed to polygenic SD (PSD), in which many genes at different loci interact to produce the sexual phenotype. Here we examine claims for PSD in vertebrates, finding that most constitute transient states during sex chromosome turnover, or aberrant systems in species hybrids. To avoid confusion about terminology, we propose a consistent nomenclature for genetic SD systems., Competing Interests: Declaration of interests No interests are declared., (Published by Elsevier Ltd.)

Considerations of the impact climate change has on reptiles are typically focused on habitat change or loss, range shifts and skewed sex ratios in species with temperature-dependent sex determination. Here, we show that incubation temperature alters stripe number and head colouration of hatchling American alligators (Alligator mississippiensis). Animals incubated at higher temperatures (33.5°C) had, on average, one more stripe than those at lower temperatures (29.5°C), and also had significantly lighter heads. These patterns were not affected by estradiol-induced sex reversal, suggesting independence from hatchling sex. Therefore, increases in nest temperatures as a result of climate change have the potential to alter pigmentation patterning, which may have implications for offspring fitness., Competing Interests: Competing interests This report was prepared as an account of work sponsored by an agency of the United States government. Neither the United States government nor any agency thereof, nor any of their employees, makes any warranty, express or implied, or assumes any legal liability or responsibility for the accuracy, completeness or usefulness of any information, apparatus, product or process disclosed. Reference herein to any specific commercial product, process or service by trade name, trademark or manufacturer, or otherwise does not necessarily constitute or imply its endorsement, recommendation, or favouring by the United States government or any agency thereof. The view and opinions of authors expressed herein do not necessarily state or reflect those of the United States government or any agency thereof., (© 2023. Published by The Company of Biologists Ltd.)

Background: The American alligator (Alligator mississippiensis) displays temperature-dependent sex determination (TSD), in which incubation temperature during embryonic development determines the sexual fate of the individual. However, the molecular mechanisms governing this process remain a mystery, including the influence of initial environmental temperature on the comprehensive gonadal gene expression patterns occurring during TSD. Results: Our characterization of transcriptomes during alligator TSD allowed us to identify novel candidate genes involved in TSD initiation. High-throughput RNA sequencing (RNA-seq) was performed on gonads collected from A. mississippiensis embryos incubated at both a male and a female producing temperature (33.5 °C and 30 °C, respectively) in a time series during sexual development. RNA-seq yielded 375.2 million paired-end reads, which were mapped and assembled, and used to characterize differential gene expression. Changes in the transcriptome occurring as a function of both development and sexual differentiation were extensively profiled. Forty-one differentially expressed genes were detected in response to incubation at male producing temperature, and included genes such as Wnt signaling factor WNT11, histone demethylase KDM6B, and transcription factor C/EBPA. Furthermore, comparative analysis of development- and sex-dependent differential gene expression revealed 230 candidate genes involved in alligator sex determination and differentiation, and early details of the suspected male-fate commitment were profiled. We also discovered sexually dimorphic expression of uncharacterized ncRNAs and other novel elements, such as unique expression patterns of HEMGN and ARX. Twenty-five of the differentially expressed genes identified in our analysis were putative transcriptional regulators, among which were MYBL2, MYCL, and HOXC10, in addition to conventional sex differentiation genes such as SOX9, and FOXL2. Inferred gene regulatory network was constructed, and the gene-gene and temperature-gene interactions were predicted. Conclusions: Gonadal global gene expression kinetics during sex determination has been extensively profiled for the first time in a TSD species. These findings provide insights into the genetic framework underlying TSD, and expand our current understanding of the developmental fate pathways during vertebrate sex determination. [ABSTRACT FROM AUTHOR]

Organisms become adapted to their environment by evolving through natural selection, a process that generally transpires over many generations. Currently, anthropogenically driven environmental changes are occurring orders of magnitude faster than they did prior to human influence, which could potentially outpace the ability of some organisms to adapt. Here, we focus on traits associated with temperature-dependent sex determination (TSD), a classic polyphenism, in a model turtle species to address the evolutionary potential of species with TSD to respond to rapid climate change. We show, first, that sex-ratio outcomes in species with TSD are sensitive to climatic variation. We then identify the evolutionary potential, in terms of heritability, of TSD and quantify the evolutionary potential of 3 key traits involved in TSD: pivotal temperature, maternal nest-site choice, and nesting phenology. We find that these traits display different patterns of adaptive potential: pivotal temperature exhibits moderate heritable variation, whereas nestsite choice and nesting phenology, with considerable phenotypic plasticity, have only modest evolutionary potential to alter sex ratios. Therefore, the most likely response of species with TSD to anthropogenically induced climate change may be a combination of microevolution in thermal sensitivity of the sex-determining pathway and of plasticity in maternal nesting behavior. [ABSTRACT FROM AUTHOR]

Temperature sex determining species offer a model for investigating how environmental cues become integrated to the regulation of patterning genes and growth, among bipotential gonads. Manipulation of steroid hormones has revealed the important role of aromatase in the regulation of the estrogen levels involved in temperature-dependent sex determination. Estradiol treatment counteracts the effect of male-promoting temperature, but the resulting ovarian developmental pattern differs from that manifested with the female-promoting temperature. Hypoplastic gonads have been reported among estradiol-treated turtles; however the estradiol effect on gonadal size has not been examined. Here we focused on the sea turtle Lepidochelys olivacea , which develops hypoplastic gonads with estradiol treatment. We studied the effect of estradiol on cell proliferation and on candidate genes involved in ovarian pattern. We found this effect is organ specific, causing a dramatic reduction in gonadal cell proliferation during the temperature-sensitive period. Although the incipient gonads resembled tiny ovaries, remodeling of the medullary cords and down-regulation of testicular factor Sox9 were considerably delayed. Contrastingly, with ovarian promoting temperature as a cue, exogenous estradiol induced the up-regulation of the ovary factor FoxL2, prior to the expression of aromatase. The strong expression of estrogen receptor alpha at the time of treatment suggests that it mediates estradiol effects. Overall results indicate that estradiol levels required for gonadal growth and to establish the female genetic network are delicately regulated by temperature. [ABSTRACT FROM AUTHOR]

In oviparous species, preovulatory maternal effects such as investment in yolk quantity and composition or levels of yolk steroid hormones might potentially affect hatchling body sizes, growth rates, performance, and sex in species with temperature-dependent sex determination. We investigated the effects of egg mass and levels of testosterone and 17β-estradiol in the yolk on the phenotypes of hatchling Magdalena River Turtles ( Podocnemis lewyana) in three populations in the Magdalena River drainage of northern Colombia. We hypothesized that, under homogeneous incubation conditions, the differences documented among clutches would be attributable to familial effects, caused by either heritable genetic factors or preovulatory maternal effects. Eggs were incubated artificially at the pivotal temperature for the species (33.4°C) and hatchlings were reared in the laboratory for 2 mo. Two eggs from each clutch were tested for testosterone and 17β-estradiol levels. Sex ratios, hatchling size, mass, and righting times varied both between clutches within a site and between populations. Egg mass was positively related to hatchling body size and mass, both at hatching and at 2 mo of age. Levels of 17β-estradiol concentrations influenced hatching success rates and incubation periods. Sex ratios were marginally related to 17β-estradiol levels, with clutches and sites with higher levels tending to produce fewer males. Our results provide evidence that preovulatory maternal effects play a role in influencing phenotypic attributes related to hatchling survivorship such as size, growth rate, and performance, and also seem to interact with incubation temperature to determine the sex of each individual and the sex ratio of the clutch, which has implications for both parental and offspring fitness. [ABSTRACT FROM AUTHOR]

Temperature-dependent sex determination (TSD) is the predominant form of environmental sex determination (ESD) in reptiles, but the adaptive significance of TSD in this group remains unclear. Additionally, the viability of species with TSD may be compromised as climate gets warmer. We simulated population responses in a turtle with TSD to increasing nest temperatures and compared the results to those of a virtual population with genotypic sex determination (GSD) and fixed sex ratios. Then, we assessed the effectiveness of TSD as a mechanism to maintain populations under climate change scenarios. TSD populations were more resilient to increased nest temperatures and mitigated the negative effects of high temperatures by increasing production of female offspring and therefore, future fecundity. That buffered the negative effect of temperature on the population growth. TSD provides an evolutionary advantage to sea turtles. However, this mechanism was only effective over a range of temperatures and will become inefficient as temperatures rise to levels projected by current climate change models. Projected global warming threatens survival of sea turtles, and the IPCC high gas concentration scenario may result in extirpation of the studied population in 50 years. [ABSTRACT FROM AUTHOR]

Sex in many reptiles is determined by temperature during development (temperature-dependent sex determination; TSD) rather than by an individual's genotype (genotypic sex determination; GSD). TSD has numerous ecological and evolutionary implications, and whether or not species display TSD is of conservation concern because substantial habitat and climate change have the potential to skew sex ratios. It is therefore important to describe the means by which sex is determined in reptile species. To date, the sex determination mechanism is not known for any species within the Anguidae, a diverse lizard family that is globally distributed. I used controlled incubation experiments to test the hypothesis that Southern Alligator Lizards ( Elgaria multicarinata) display TSD. Offspring sex was assessed by direct examination of the gonads after incubation in one of five constant-temperature treatments (24°, 26°, 28°, 30°, and 32°C). Developmental temperature did not affect offspring sex, indicating that E. multicarinata does not display a strict form of TSD. Rather, my results indicate that E. multicarinata displays GSD, as is most common in vertebrates, although I cannot rule out the possibility that sex determination is somewhat thermally sensitive. Further work with additional samples and other anguid species is clearly needed. [ABSTRACT FROM AUTHOR]

The article focuses on the care and breeding of fringe-toed lizards Phrynosomatidae at the Phoenix Zoo, U.S. Topics discussed include captive husbandry of the lizard under warm temperature; breeding efforts by female from March 3, 2013 and September 22, 2013; laying of eggs on the bottom of the nest boxes; incubation of eggs and determination of sex of hatchling over half a year in age and impact of temperature to sex determination of Uma.