scholarly journals Improved genome assembly of American alligator genome reveals conserved architecture of estrogen signaling

2016 ◽  
Author(s):  
Edward S. Rice ◽  
Satomi Kohno ◽  
John St. John ◽  
Son Pham ◽  
Jonathan Howard ◽  
...  

AbstractThe American alligator, Alligator mississippiensis, like all crocodilians, has temperature-dependent sex determination, in which the sex of an embryo is determined by the incubation temperature of the egg during a critical period of development. The lack of genetic differences between male and female alligators leaves open the question of how the genes responsible for sex determination and differentiation are regulated. One insight into this question comes from the fact that exposing an embryo incubated at male-producing temperature to estrogen causes it to develop ovaries. Because estrogen response elements are known to regulate genes over long distances, a contiguous genome assembly is crucial for predicting and understanding its impact.We present an improved assembly of the American alligator genome, scaffolded with in vitro proximity ligation (Chicago) data. We use this assembly to scaffold two other crocodilian genomes based on synteny. We perform RNA sequencing of tissues from American alligator embryos to find genes that are differentially expressed between embryos incubated at male-versus female-producing temperature. Finally, we use the improved contiguity of our assembly along with the current model of CTCF-mediated chromatin looping to predict regions of the genome likely to contain estrogen-responsive genes. We find that these regions are significantly enriched for genes with female-biased expression in developing gonads after the critical period during which sex is determined by incubation temperature. We thus conclude that estrogen signaling is a major driver of female-biased gene expression in the post-temperature sensitive period gonads.

2007 ◽  
Vol 28 (1) ◽  
pp. 123-128 ◽  
Author(s):  
Carlos Piña ◽  
Alejandro Larriera ◽  
Pablo Siroski ◽  
Luciano Verdade ◽  
Valentine Lance

AbstractAll crocodiles studied to date exhibit temperature-dependent sex determination. During the many weeks from egg laying to hatch there is a period of 10 to 15 d in the middle third of incubation (in the American alligator) during which the sex of the embryo is irreversibly fixed, referred to as the temperature-sensitive period or TSP. In this work we investigated the TSP in Caiman latirostris eggs incubated at female-inducing and male-inducing temperatures (29° C and 33° C respectively) by switching eggs from 29° C to 33° C and vice versa at timed interval throughout incubation. Compared to Alligator mississippiensis the duration of TSP was longer, and the onset of TSP was at an earlier stage of incubation.


1996 ◽  
Vol 149 (3) ◽  
pp. 457-463 ◽  
Author(s):  
D Crews ◽  
A R Cantú ◽  
J M Bergeron

Abstract This study addressed the hypothesis that, in the red-eared slider turtle, Trachemys scripta, non-aromatizable androgens are the physiological equivalent of temperature in determining male development. In the first experiment, eggs were treated in the middle of the temperature-sensitive period with 1·0 or 10·0 μg androsterone, 5α-dihydrotestosterone, 3α-androstanediol, or 3β-androstanediol, while at an all-male, male-biased, or one of two female-biased incubation temperatures. In the second experiment, eggs were treated with the same dosages of dihydrotestosterone at different stages of embryonic development while at a male-biased, threshold, or a female-biased incubation temperature. Results of experiment one indicated that hormone-induced masculinization is specific to non-aromatizable androgens. Results of experiment two indicated that the sensitivity to dihydrotestosterone corresponds to the temperature-sensitive window during development. Further, there is a dose–response relationship but no apparent synergism between exogenous dihydrotestosterone and incubation temperature. When considered with other research, it is suggested that non-aromatizable androgens and their products are involved in the initiation of male sex determination whereas oestrogens and their aromatizable androgen precursors are involved in the initiation of female sex determination. Journal of Endocrinology (1996) 149, 457–463


1983 ◽  
Vol 10 (3) ◽  
pp. 607 ◽  
Author(s):  
GJW Webb ◽  
SC Manolis ◽  
R Buckworth

In the McKinlay River area most female C. johnstoni mature at 74-78 cm snout-vent length (SVL), and 11-14 years of age, whereas maies are about 87 cm SYL and 16-17 years of age. Adult sex ratios in the field are lM:3-4F. There is a well defined, brief nesting period in late August and early September, and the nests are typical 'hole' nests excavated in sand or other friable substrates close to permanent water. Mean clutch size is 13.2 � 3.2 eggs (� SD) and mean egg dimensions are: length 6.64 � 0.26 cm, breadth 4.19 � 0.19 cm and weight 68.2 � 8.0 g. There is a variable but significant increase in egg size with clutch size, and large clutches of large eggs tend to be laid earlier in the nesting season than small clutches of small eggs. From 2 to 4% of eggs are infertile. Nest temperatures show considerable daily variation and baseline levels differ between shaded and exposed nest sites. Incubation times are temperaturedependent and range from 9 to 14 weeks. Females excavate nests at hatching time, but attend the nests little, ifat all, during incubation. Hatchlings are 11.2 � 0.5 cm SVL and weigh 42.0 � 6.1 g. Formulae for predicting egg and hatchling dimensions from each other are presented. A preliminary method for aging C. johnstoni embryos is described. Artificial incubation at 26�, 30� and 34�C, respectively, resulted in 0, 63% and 21% survival; at 26 and 34�C physically deformed embryos were common. Hatching success is correlated with the age at which eggs are transported to incubators. Sex determinantion is influenced by incubation temperature, and at temperatures we tested between 26 and 34�C, females predominated; males were produced at 31-32�C. Histological examination of females from high-temperature incubation (34�C) indicated slight hermaphroditic tendencies in two of 35 animals examined; their status remains to be clarified. The temperature-sensitive period for sex determination appears to be between 20 and 57 days of age (30�C equivalent ages), but this may well vary with incubation temperature, as in Alligator mississippiensis. Egg losses due to predators (particular varanid lizards) were estimated as 64%, although they may have been increased by our interference with nests. Eggs are also lost to flooding and overheating, and a 60-70% annual egg mortality may be common. In one experiment, hatchling mortality was estimated at 98% within the first year. An egg and/or hatchling harvest balanced by a proportional return to the wild of raised 1-year-olds (5% and 10% of eggs and hatchlings collected respectively) is a potential strategy through which sustained-yield harvesting could be introduced into a conservation-management program.


2016 ◽  
Vol 236 ◽  
pp. 63-69 ◽  
Author(s):  
Christopher M. Murray ◽  
Michael Easter ◽  
Mark Merchant ◽  
Justin L. Rheubert ◽  
Kelly A. Wilson ◽  
...  

The various patterns of environmental sex determination in squamates, chelonians and crocodilians are described. High temperatures produce males in lizards and crocodiles but females in chelonians. Original experiments on the effects of incubation at 30 °C (100% females) or 33 °C (100% males) on development in Alligator mississippiensis are described. These include an investigation of the effect of exposing embryos briefly to a different incubation temperature on the sex ratio at hatching, and a study of the effects of 30 °C and 33 °C on growth and development of alligator embryos and gonads. A 7-day pulse of one temperature on the background of another was insufficient to alter the sex ratio dramatically. Incubation at 33 °C increased the rate of growth and development of alligator embryos. In particular, differentiation of the gonad at 33 °C was enhanced compared with 30 °C. A hypothesis is developed to explain the mechanism of temperature-dependent sex determination (TSD) in crocodilians. The processes of primary sex differentiation are considered to involve exposure to a dose of some male-determining factor during a specific quantum of developmental time during early incubation. The gene that encodes for the male- determining factor is considered to have an optimum temperature (33 °C). Any change in the temperature affects the expression o f this gene and affects the dose or quantum embryos are exposed to. In these cases there is production of females by default. The phylogenetic implications of TSD for crocodilians, and reptiles in particular, are related to the life history of the animal from conception to sexual maturity. Those animals that develop under optimal conditions grow fastest and largest and become male. A general association between the size of an animal and its sex is proposed for several types of vertebrate.


1986 ◽  
Vol 43 (3) ◽  
pp. 514-520 ◽  
Author(s):  
David O. Conover ◽  
Mark H. Fleisher

We investigated the timing and duration of the temperature-sensitive period of development in the Atlantic silverside, Menidia menidia, by measuring the sex ratio in groups of larvae shifted reciprocally among low and high temperatures as development progressed. The sensitive period of sex determination was dependent on body size, rather than age, and occurred during the middle to later stages of larval development when fish were between 8 and 21 mm in total length. Sex determination was irreversibly fixed earlier in development (i.e. at 15 mm in length) at higher than at lower temperatures. Higher, male-producing temperatures therefore appear to more strongly effect sex determination than do lower, female-producing temperatures. Temperature prior to, or after, the sensitive period had no measurable effect on sex ratio. From histological examination we suggest that gonad differentiation coincided with the end of the temperature-sensitive period. The timing and duration of the temperature-sensitive period in M. menidia is generally similar to the hormone-sensitive period identified in other fishes. Both phenomena reveal a specific developmental period during which the course of primary sex differentiation in fishes is labile.


Endocrinology ◽  
2015 ◽  
Vol 156 (5) ◽  
pp. 1887-1899 ◽  
Author(s):  
Satomi Kohno ◽  
Melissa C. Bernhard ◽  
Yoshinao Katsu ◽  
Jianguo Zhu ◽  
Teresa A. Bryan ◽  
...  

All crocodilians and many turtles exhibit temperature-dependent sex determination where the temperature of the incubated egg, during a thermo-sensitive period (TSP), determines the sex of the offspring. Estrogens play a critical role in sex determination in crocodilians and turtles, as it likely does in most nonmammalian vertebrates. Indeed, administration of estrogens during the TSP induces male to female sex reversal at a male-producing temperature (MPT). However, it is not clear how estrogens override the influence of temperature during sex determination in these species. Most vertebrates have 2 forms of nuclear estrogen receptor (ESR): ESR1 (ERα) and ESR2 (ERβ). However, there is no direct evidence concerning which ESR is involved in sex determination, because a specific agonist or antagonist for each ESR has not been tested in nonmammalian species. We identified specific pharmaceutical agonists for each ESR using an in vitro transactivation assay employing American alligator ESR1 and ESR2; these were 4,4′,4′’-(4-propyl-[1H]-pyrazole-1,3,5-triyl)trisphenol (PPT) and 7-bromo-2-(4-hydroxyphenyl)-1,3-benzoxazol-5-ol (WAY 200070), respectively. Alligator eggs were exposed to PPT or WAY 200070 at a MPT just before the TSP, and their sex was examined at the last stage of embryonic development. Estradiol-17β and PPT, but not WAY 200070, induced sex reversal at a MPT. PPT-exposed embryos exposed to the highest dose (5.0 μg/g egg weight) exhibited enlargement and advanced differentiation of the Müllerian duct. These results indicate that ESR1 is likely the principal ESR involved in sex reversal as well as embryonic Müllerian duct survival and growth in American alligators.


Genetics ◽  
1993 ◽  
Vol 135 (3) ◽  
pp. 831-841 ◽  
Author(s):  
Y Hiraizumi

Abstract Previous work has shown that the direction of segregation distortion in the SD (Segregation Distorter) system in Drosophila melanogaster can sometimes be reversed, but this was found only with rather weak distorters and the effect was not large. The present study reports large negative segregation distortion in a strong distorter, SD-72 chromosome. In the presence of a specific X chromosome, supp-X(SD), the proportion, k, of SD-72 chromosomes recovered from the SD-72/cn bw males ranges from 0.99 at 20 degrees to 0.11 at 28.5 degrees, whereas with a standard-X chromosome, k ranges from 0.99 to 0.95 for the same temperature range. The temperature-sensitive period is during spermiogenesis. Using a mating system in which the sperm supply is nearly exhausted, it was shown that the negative distortion at high temperatures is due to an absolute reduction in the number of SD-72 chromosomes and an absolute increase in the number of cn bw chromosomes recovered. After adjusting for non-SD-related temperature effects, the amount of decrease in the number of SD-72 progeny is nearly the same as the amount of increase in the number of cn bw progeny, suggesting that the dysfunction switches from a spermatid carrying one homolog to one carrying the other. Negative distortion requires a radical revision of current hypotheses for the mechanism of segregation distortion and a possible modification of the current model is suggested, based on differential recovery of dysfunction in the two homologs during spermiogenesis.


1994 ◽  
Vol 63 (2) ◽  
pp. 117-127 ◽  
Author(s):  
Marc Girondot ◽  
Patrick Zaborski ◽  
Jean Servan ◽  
Claude Pieau

SummaryIn many reptiles, sex determination is temperature-sensitive. This phenomenon has been shown to take place in the laboratory as well as in nature, but its effect on natural populations remains questionable. In the turtle Emys orbicularis, the effects of temperature override a weak mechanism of genetic sex determination which is revealed in incubation at pivotal temperature. At this temperature, the sexual phenotype is concordant with the expression of the serologically defined H-Y antigen (H-Ys) in non-gonadal tissues; males are H-Ys negative (H-Y−) whereas females are H-Ys positive (H-Y+). To estimate the importance of sexual inversion (sexual phenotype and H-Ys expression discordant) in populations of Brenne (France), the frequencies of male and female sexual phenotypes among H-Ys phenotypes were determined. The frequencies of sex reversed individuals are low, only 6 % of phenotypic females being H-Y− and 11 % of phenotypic males being H-Y+. According to these data, two theoretical models have been constructed to estimate the contribution to sex determination of individuals in relation to their genotype. The first model excludes any influence of incubation temperature and sexual phenotype on the fitness of individuals. The second one considers that these parameters influence fitness because this model has been previously shown to favour environmental sex determination. In both models, it appears that sex determination can be viewed as genotypic and monogenic with some individuals sexually inverted by theaction of temperature. One category of homozygous animals differentiates mainly into one sex, and the heterozygous animals differentiate mainly into the other sex. The second category of homozygotes has a low frequency in the populations and can differentiate as male or female without high constraint. Then it is estimated that in Brenne approximately 83% of the eggs are incubated in conditions allowing the genetic component to influence sex determination.


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