[1].The Effect of the Cyp19a1 Gene Methylation Modification on Temperature-dependent Sex Determination of Reeves’ Turtle (Mauremys reevesii)[J].Asian Herpetological Research,2017,8(3):213-220.[doi:10.16373/j.cnki.ahr.170021]
 Wenxiu RU,**,Liushuai HUA,et al.The Effect of the Cyp19a1 Gene Methylation Modification on Temperature-dependent Sex Determination of Reeves’ Turtle (Mauremys reevesii)[J].Asian Herpetological Reserch(AHR),2017,8(3):213-220.[doi:10.16373/j.cnki.ahr.170021]

The Effect of the Cyp19a1 Gene Methylation Modification on Temperature-dependent Sex Determination of Reeves’ Turtle (Mauremys reevesii)()

Asian Herpetological Research[ISSN:2095-0357/CN:51-1735/Q]



The Effect of the Cyp19a1 Gene Methylation Modification on Temperature-dependent Sex Determination of Reeves’ Turtle (Mauremys reevesii)
Wenxiu RU1 2** Liushuai HUA2 3** Yufeng WEI2 Weiye LI2 Dainan CAO2 Yan GE2 Hong CHEN1 Xianyong LAN1* and Shiping GONG2*
1 Shaanxi Key Laboratory of Molecular Biology for Agriculture, College of Animal Science and Technology, Northwest A&F University, Yangling 712100, Shaanxi, China
2 Guangdong Key Laboratory of Animal Conservation and Resource Utilization, Guangdong Public Laboratory of Wild Animal Conservation and Utilization, Guangdong Institute of Applied Biological Resources, Guangzhou 510260, China
3 Henan Key Laboratory of Farm Animal Breeding and Nutritional Regulation, Institute of Animal Husbandry and Veterinary Science, Henan Academy of Agricultural Sciences, Zhengzhou 450002, Henan, China
Reeves’ turtle Mauremys reevesii temperature-dependent sex determination (TSD) cyp19a1 gene methylation
Temperature-dependent sex determination (TSD) is a type of environmental sex determination in which the sex of the embryos depends on the ambient temperature; however, the molecular mechanisms governing this process remain unknown. Aromatase, encoded by the cyp19a1 gene, which converts androgens into estrogens in animals, was considered to be the key gene for TSD. In this study, the 5’-flanking region of the cyp19a1 gene in Reeves’ turtle (Mauremys reevesii) was cloned, and the promoter region was identified using the luciferase reporter assay. Then the eggs of Reeves’ turtle were incubated at different temperatures (26°C: male-biased temperature; 29°C: non-sex-biased temperature and 32°C: female-biased temperature). During the thermosensitive period, the adrenal kidney gonad complexes (AKG) were sampled. DNA methylation analysis of the AKG samples showed that the promoter region of the cyp19a1 gene was significantly de-methylated in the female-biased temperature regime (P<0.01). Quantitative analysis of the cyp19a1 gene and estrogen by qPCR and Elisa assay showed that the expression level of the cyp19a1 gene and estrogen content were both upregulated significantly at the female-biased temperature (P<0.01). These results indicated that the de-methylation response of the cyp19a1 gene to incubation temperature, especially at the female-biased temperature, could lead to temperature-specific expression of aromatase and increased estrogen levels, which may further determine gonadal development in Reeves’ turtle. These findings provide insights into the genetic mechanisms underlying TSD.


Bieser K. L, Wibbels T. 2014. Chronology, Magnitude and Duration of Expression of Putative Sex-Determining/Differentiation Genes in a Turtle with Temperature-Dependent Sex Determination. Sex Dev, 8: 364–375
Bull J. 1980. Sex determination in reptiles. Q Rev Biol, 55: 3–21
Caruso C. C., Breton T. S., Berlinsky D. L. 2015. The effects of temperature on ovarian aromatase (cyp19a1a) expression and sex differentiation in summer flounder (Paralichthys dentatus). Fish Physiol Biochem, 42: 795–805
Crews D. 1996. Temperature-dependent sex determination: the interplay of steroid hormones and temperature. Zool Sci, 13: 1–13
Crews D., Bull J. J., Wibbels T. 1991. Estrogen and sex reversal in turtles: a dose-dependent phenomenon. Gen Comp Endocr, 81: 357–364
Díaz-Hernández V., Marmolejo-Valencia A., Merchant-Larios H. 2015. Exogenous estradiol alters gonadal growth and timing of temperature sex determination in gonads of sea turtle. Dev biol, 408: 79–89
Du W. G., Hu L. J., Lu J. L., Zhu L. J. 2007. Effects of incubation temperature on embryonic development rate, sex ratio and post-hatching growth in the Chinese three-keeled pond turtle, Chinemys reevesii. Aquaculture, 272: 747–753
Du W. G., Sun B. J. 2012.The phenomenon and mechanism of temperature-dependent sex determination in reptiles. Bull Biol, 47: 1–5 (In Chinese with English abstract)
Greenbaum E. 2002. A standardized series of embryonic stages for the emydid turtle Trachemys scripta. Can J Zool, 80: 1350–1370
He B., Shi H. T., Liao G. Q. 2009. Temperature-dependent sex determination in turtles. Chinese J Zool, 5: 147–152 (In Chinese with English abstract)
Hou L. 1985. Sex determination by temperature for incubation in Chinemys reevesii. Acta Herpetol Sin, 4: 130
Jaenisch R., Bird A. 2003. Epigenetic regulation of gene expression: how the genome integrates intrinsic and environmental signals. Nat genet, 33: 245–254
Li S. 2002. Study on the temperature-dependent sexual differentiation in turtle (Chinemys reevesii). Thesis, Hunan Normal University, China (In Chinese with English abstract)
Matsumoto Y., Buemio A., Chu R., Vafaee M., Crews D. 2013. Epigenetic control of gonadal aromatase cyp19a1 in temperature-dependent sex determination of red-eared slider turtles. PLoS One, 8: e63599
Matsumoto Y., Hannigan B., Crews D. 2014. Embryonic PCB exposure alters phenotypic, genetic, and epigenetic profiles in turtle sex determination, a biomarker of environmental contaminatio. Endocrinology, 155(11): 4168–4177
Nakamoto M., Wang D. S., Suzuki A., Matsuda M., Nagahama Y., Shibata N. 2007. Dax1 suppresses P450arom expression in medaka ovarian follicles. Mol Reprod Dev, 74: 1239–1246
Navarro-Martín L., Vi?as J., Ribas L., Díaz N., Gutiérrez A., Di Croce L., Piferrer F. 2011. DNA methylation of the gonadal aromatase (cyp19a) promoter is involved in temperature-dependent sex ratio shifts in the European sea bass. PLoS Genet, 7: e1002447
Oshima Y., Kato T., Wang D., Murakami T , Matsuda Y., Nagahama Y., Nakamura M. 2006. Promoter activity and chromosomal location of the Rana rugosa P450 aromatase (CYP19) gene. Zool Sci, 23: 79–85
Ospina-?lvarez N., Piferrer F. 2008. Temperature-dependent sex determination in fish revisited: Prevalence, a single sex ratio response pattern, and possible effects of climate change. PLoS One, 3: e2837
Parrott B. B., Kohno S., Cloy-McCoy J. A., Guillette L. J. 2013. Differential Incubation Temperatures Result in Dimorphic DNA Methylation Patterning of the SOX9 and Aromatase Promoters in Gonads of Alligator (Alligator mississippiensis) Embryos. Biol of Reprod, 90: 1–11
Pieau C., Mignot T. M., Dorizzi M., Guichard A. 1982. Gonadal steroid levels in the turtle Emys orbicularis L.: A preliminary study in embryos, hatchlings, and young as a function of the incubation temperature of eggs. Gen Comp Endocr, 47: 392–398
Piferrer F. 2013. Epigenetics of sex determination and gonadogenesis. Dev Dynam, 242: 360–370
Ramsey M., Shoemaker C., Crews D. 2007. Gonadal expression of Sf1 and aromatase during sex determination in the red-eared slider turtle (Trachemys scripta), a reptile with temperature-dependent sex determination. Differentiation, 75: 978–991
Razin A., Riggs A. D. 1980. DNA methylation and gene function. Science, 210: 604–610
Shoemaker C. M., Queen J., Crews D. 2007. Response of candidate sex-determining genes to changes in temperature reveals their involvement in the molecular network underlying temperature-dependent sex determination. Mol Endocrinol, 21: 2750–2763
Si Y., Ding Y., He F., Wen H., Li J., Zhao J., Huang Z. 2016. DNA methylation level of cyp19a1a and Foxl2 gene related to their expression patterns and reproduction traits during ovary development stages of Japanese flounder (Paralichthys olivaceus). Gene, 575: 321–330
Simpson E. R., Mahendroo M. S., Means G. D., Kilgore M. W., Hinshelwood M. M., Graham-Lorence S., Amarneh B., Ito Y., Fisher C. R., Michael M. D.
1994. Aromatase Cytochrome P450, The Enzyme Responsible for Estrogen Biosynthesis. Endocr rev, 15: 342–355
Tan L. J, Liu J., Chen S. Q. 2001. The study of embryonic development in turtle(Chinemys reevesii). Acta Hydrobiol Sin, 6: 605–612 (In Chinese with English abstract)
Vandeputte M., Dupont-Nivet M., Chavanne H., Chatain B. 2007. A polygenic hypothesis for sex determination in the European sea bass Dicentrarchus labrax. Genetics, 176: 1049–1057
Yang Z. B. 2011. Observation on the morphology of embryonic development of Chinese Striped-neck Turtle, Mauremys sinensis. J Anhui Agr Sci, 33: 20501–20505 (In Chinese with English abstract).
Wang D. S., Kobayashi T., Zhou L. Y., Paul-Prasanth B., Ijiri S., Sakai F., Okubo K., Morohashi K., Nagahama Y. 2007. Foxl2 up-regulates aromatase gene transcription in a female-specific manner by binding to the promoter as well as interacting with ad4 binding protein/steroidogenic factor 1. Mol Endocrinol, 21: 712–725
Wang D. S., Zhou L. Y., Kobayashi T., Matsuda M., Shibata Y., Sakai F., Nagahama Y. 2010. Doublesex- and Mab-3-related transcription factor-1 repression of aromatase transcription, a possible mechanism favoring the male pathway in tilapia. Endocrinology, 151: 1331–1340

更新日期/Last Update: 2017-09-25