Imprinted genes play significant roles in the regulation of fetal growth, development, function of the placenta and postnatal behavior in mammals, but little is known in pigs. In order to investigate the imprinting st...Imprinted genes play significant roles in the regulation of fetal growth, development, function of the placenta and postnatal behavior in mammals, but little is known in pigs. In order to investigate the imprinting status of porcine retro-transposon like 1 （RTL1） and type 3 iodothyronine deiodinase （DIO3） genes, DNA or RNA samples of the parents and F1 animals, generated with reciprocal crosses between Large White and Meishan breeds, were isolated, and analyzed by reverse transcription polymerase chain reaction restriction fragment length polymorphism （RT-PCR-RFLP）. The results demonstrated that the RTL1 gene was paternally expressed in 10 tissues, such as the skeletal muscle, heart, spleen, liver, kidney, lung, stomach, fat, small intestine and brain, and D103 gene exhibited paternal expression in the skeletal muscle, heart, spleen, lung, stomach, and brain, in 2-month-old pigs. The association of RTL1 and DI03 with carcass traits was further analyzed in the F2 population of Large White×Meishan pigs. The statistical results showed that the R TL1 A1101G polymorphism （EU781029） was significantly associated with lean meat percentage （LMP） and fat meat percentage （FMP） （P〈0.05）, while the D103 A744C polymorphism （AY533208） was not significantly associated with any carcass traits. These results indicate that the imprinting status of RTL1 and DIO3 is well kept across the mammalian species, and porcine RTL1 may have important roles in muscle growth and fat deposition.展开更多
AIM:To examine the imprinted Dlk1-Dio3 locus in pluripotent embryonic stem(ES)cell/fibroblast hybrid cells.METHODS:Gtl2,Rian,and Mirg mRNA expression in mouse pluripotent ES cell/fibroblast hybrid cells was examined b...AIM:To examine the imprinted Dlk1-Dio3 locus in pluripotent embryonic stem(ES)cell/fibroblast hybrid cells.METHODS:Gtl2,Rian,and Mirg mRNA expression in mouse pluripotent ES cell/fibroblast hybrid cells was examined by real-time reverse transcription-polymerase chain reaction.Pyrosequencing and bisulfate sequencing were used to determine the DNA methylation level of the Dlk1-Dio3 locus imprinting control region. RESULTS:The selected hybrid clones had a near-tetraploid karyotype and were highly pluripotent judging from their capacity to generate chimeric embryos and adult chimeras.Our data clearly demonstrate that Gtl2,Rian,and Mirg,which are imprinted genes within the Dlk1-Dio3 locus,are active in all examined ES cell/fibroblast hybrid clones.In spite of interclonal variability,the expression of the imprinted genes is comparable to that of ES cells and fibroblasts.Quantitative analysis of the DNA methylation status of the intergenic differentially methylated region(IG DMR)within the Dlk1-Dio3 locus by pyrosequencing and bisulfite sequencing clearly showed that the DNA methylation status of the imprinted region in the tested hybrid clones was comparable to that of both ES cells and fibroblasts.CONCLUSION:Reprogramming process in a hybrid cell system is achieved without marked alteration of the imprinted Dlk1-Dio3 locus.展开更多
Photoperiod,the length of daylight,has a significant impact on the physiological characteristics of seasonal breeding animals,including their somatic and gonadal development.In rodents,expression of deiodinase type II...Photoperiod,the length of daylight,has a significant impact on the physiological characteristics of seasonal breeding animals,including their somatic and gonadal development.In rodents,expression of deiodinase type II(Dio2)and III(Dio3)in the hypothalamus is crucial for responding to photoperiodic signals.However,research on the photoperiodism of hypothalamic gene expression and the corresponding regulatory mechanism in Brandt’s voles living in the Mongolian steppes is limited.In this study,we gradually changed day length patterns to simulate spring(increasing long photoperiod,ILP)and autumn(decreasing short photoperiod,DSP).We compared the somatic and gonadal development of voles born under ILP and DSP and the expression patterns of five reproductionrelated genes in the hypothalamus of young voles.The results showed that DSP significantly inhibited somatic and gonadal development in both female and male offspring.Compared with ILP,Dio3 expression was significantly upregulated in the hypothalamus under DSP conditions and remained elevated until postnatal week 8 in both males and females.However,there was no significant difference in the methylation levels of the proximal promoter region of Dio3 between ILP and DSP,suggesting that methylation in the proximal promoter region may not be involved in regulating the expression of Dio3.These findings suggest that hypothalamic expression of Dio3 plays a key role in the photoperiodic regulation of gonadal activity in Brandt’s voles.However,it appears that CpGs methylation in the promoter region is not the main mechanism regulating Dio3 expression.展开更多
基金supported by the National Natural Sci-ence Foundation of China (30571331)the China Postdoctoral Science Foundation (2005038161)
文摘Imprinted genes play significant roles in the regulation of fetal growth, development, function of the placenta and postnatal behavior in mammals, but little is known in pigs. In order to investigate the imprinting status of porcine retro-transposon like 1 （RTL1） and type 3 iodothyronine deiodinase （DIO3） genes, DNA or RNA samples of the parents and F1 animals, generated with reciprocal crosses between Large White and Meishan breeds, were isolated, and analyzed by reverse transcription polymerase chain reaction restriction fragment length polymorphism （RT-PCR-RFLP）. The results demonstrated that the RTL1 gene was paternally expressed in 10 tissues, such as the skeletal muscle, heart, spleen, liver, kidney, lung, stomach, fat, small intestine and brain, and D103 gene exhibited paternal expression in the skeletal muscle, heart, spleen, lung, stomach, and brain, in 2-month-old pigs. The association of RTL1 and DI03 with carcass traits was further analyzed in the F2 population of Large White×Meishan pigs. The statistical results showed that the R TL1 A1101G polymorphism （EU781029） was significantly associated with lean meat percentage （LMP） and fat meat percentage （FMP） （P〈0.05）, while the D103 A744C polymorphism （AY533208） was not significantly associated with any carcass traits. These results indicate that the imprinting status of RTL1 and DIO3 is well kept across the mammalian species, and porcine RTL1 may have important roles in muscle growth and fat deposition.
基金Supported by Grant from the Russian Academy of Sciences,Siberian Branch N48
文摘AIM:To examine the imprinted Dlk1-Dio3 locus in pluripotent embryonic stem(ES)cell/fibroblast hybrid cells.METHODS:Gtl2,Rian,and Mirg mRNA expression in mouse pluripotent ES cell/fibroblast hybrid cells was examined by real-time reverse transcription-polymerase chain reaction.Pyrosequencing and bisulfate sequencing were used to determine the DNA methylation level of the Dlk1-Dio3 locus imprinting control region. RESULTS:The selected hybrid clones had a near-tetraploid karyotype and were highly pluripotent judging from their capacity to generate chimeric embryos and adult chimeras.Our data clearly demonstrate that Gtl2,Rian,and Mirg,which are imprinted genes within the Dlk1-Dio3 locus,are active in all examined ES cell/fibroblast hybrid clones.In spite of interclonal variability,the expression of the imprinted genes is comparable to that of ES cells and fibroblasts.Quantitative analysis of the DNA methylation status of the intergenic differentially methylated region(IG DMR)within the Dlk1-Dio3 locus by pyrosequencing and bisulfite sequencing clearly showed that the DNA methylation status of the imprinted region in the tested hybrid clones was comparable to that of both ES cells and fibroblasts.CONCLUSION:Reprogramming process in a hybrid cell system is achieved without marked alteration of the imprinted Dlk1-Dio3 locus.
基金supported by the National Natural Science Foundation of China(31972284,32090022,32372571,and 31471790)the Xinjiang Tian-Chi Talents Introduction Program and the Project of Northern Agriculture and Livestock Husbandary Technical Innovation Center,Chinese Academy of Agricultural Sciencies(BFGJ2022007).
文摘Photoperiod,the length of daylight,has a significant impact on the physiological characteristics of seasonal breeding animals,including their somatic and gonadal development.In rodents,expression of deiodinase type II(Dio2)and III(Dio3)in the hypothalamus is crucial for responding to photoperiodic signals.However,research on the photoperiodism of hypothalamic gene expression and the corresponding regulatory mechanism in Brandt’s voles living in the Mongolian steppes is limited.In this study,we gradually changed day length patterns to simulate spring(increasing long photoperiod,ILP)and autumn(decreasing short photoperiod,DSP).We compared the somatic and gonadal development of voles born under ILP and DSP and the expression patterns of five reproductionrelated genes in the hypothalamus of young voles.The results showed that DSP significantly inhibited somatic and gonadal development in both female and male offspring.Compared with ILP,Dio3 expression was significantly upregulated in the hypothalamus under DSP conditions and remained elevated until postnatal week 8 in both males and females.However,there was no significant difference in the methylation levels of the proximal promoter region of Dio3 between ILP and DSP,suggesting that methylation in the proximal promoter region may not be involved in regulating the expression of Dio3.These findings suggest that hypothalamic expression of Dio3 plays a key role in the photoperiodic regulation of gonadal activity in Brandt’s voles.However,it appears that CpGs methylation in the promoter region is not the main mechanism regulating Dio3 expression.
