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 r...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＆lt;0.01）.Quantitative analysis of the cyp19a1 gene and estrogen by q PCR 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＆lt;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.展开更多
Porcine reproductive traits are characterized by low heritability, making improvement by traditional selective breeding rather difficult. Molecular breeding offers powerful approaches to overcome previous limitations ...Porcine reproductive traits are characterized by low heritability, making improvement by traditional selective breeding rather difficult. Molecular breeding offers powerful approaches to overcome previous limitations and is expected to generate economic benefits via progress in pig breeding. Cytochrome P450 family 19 subfamilyA polypeptide 1 （CYP19A1） gene is a key enzyme of estradiol biosynthesis that plays an important role in the establishment of gestation and maintenance of pregnancy. In this study, the sequence and structure characteristics of the porcine CYP19A1 gene was analyzed and expression patterns of CYP19A 1 in different tissues of adult female pigs were detected. Fourteen single-nucleotide poly- morphisms （SNPs） in the exons and introns of porcine CYP19A1 were identified and genotyped using the Sequenom MassARRAY platform, after which the allele frequency of each SNP was analyzed..The association between CYP19A1 SNPs and litter size and piglet birth weight was assessed in a crossbred pig population （n=375）. The expression pattern of CYPf9A1 revealed that it was highly expressed in the ovary, spleen, and uterus and lowly expressed in the other tissues. Moreover, one SNP, rs341891833, was significantly associated with piglet birth weight during the multiparity period （P〈0.01）. We concluded that CYP19A1 could be used as a candidate molecular marker in breeding aimed at rapid improvement of the reproductive characteristics of pigs.展开更多
基金supported financially by the National Natural Science Foundation of China(Nos.31401053 and 31471966)Guangdong Provincial Natural Science Foundation of China(No.2015A030313903)+1 种基金GDAS Special Project of Science and Technology Development(2017GDASCX-0107)the Funds for Environment Construction and Capacity Building of GDAS’Research Platform(2016GDASPT-0107)
文摘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＆lt;0.01）.Quantitative analysis of the cyp19a1 gene and estrogen by q PCR 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＆lt;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.
基金supported by the National Natural Science Foundation of China(31501931)the 948 Program of China(2011-G11)the Agricultural Science and Technology Innovation Program,China(ASTIP-IAS05)
文摘Porcine reproductive traits are characterized by low heritability, making improvement by traditional selective breeding rather difficult. Molecular breeding offers powerful approaches to overcome previous limitations and is expected to generate economic benefits via progress in pig breeding. Cytochrome P450 family 19 subfamilyA polypeptide 1 （CYP19A1） gene is a key enzyme of estradiol biosynthesis that plays an important role in the establishment of gestation and maintenance of pregnancy. In this study, the sequence and structure characteristics of the porcine CYP19A1 gene was analyzed and expression patterns of CYP19A 1 in different tissues of adult female pigs were detected. Fourteen single-nucleotide poly- morphisms （SNPs） in the exons and introns of porcine CYP19A1 were identified and genotyped using the Sequenom MassARRAY platform, after which the allele frequency of each SNP was analyzed..The association between CYP19A1 SNPs and litter size and piglet birth weight was assessed in a crossbred pig population （n=375）. The expression pattern of CYPf9A1 revealed that it was highly expressed in the ovary, spleen, and uterus and lowly expressed in the other tissues. Moreover, one SNP, rs341891833, was significantly associated with piglet birth weight during the multiparity period （P〈0.01）. We concluded that CYP19A1 could be used as a candidate molecular marker in breeding aimed at rapid improvement of the reproductive characteristics of pigs.
