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小鼠早期胚胎发育过程中的DNA去甲基化 总被引：1，自引：0，他引：1
表观遗传修饰在基因转录与表达、细胞生长与分化以及动物个体正常发育等过程中都具有重要的调控作用。表观遗传修饰发生异常,会引起机体生长发育中的各种异常。哺乳动物从精卵受精到附植前的胚胎早期发育阶段会发生重要的表观遗传重编程,主要包括DNA甲基化和组蛋白修饰。精卵受精后DNA发生主动和被动2种方式的去甲基化。本文主要综述了与DNA甲基化相关的蛋白和早期胚胎发育过程中的去甲基化机制,并对小鼠附植前胚胎发育过程中的DNA甲基化的动态变化进行了详细的论述。 相似文献
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采用成年小鼠的卵丘细胞核、胎儿成纤维细胞作核供体细胞来进行核移植,研究小鼠卵母细胞的去核程序和影响重构胚附植前发育的激活条件;随后将重构胚与供体细胞系共培养后获得囊胚阶段的小鼠重构胚,并将其移植到受体鼠体内后获得了24%克隆胚胎。结果表明,小鼠卵母细胞质能够进行重编程来支持早期的胚胎发育。 相似文献
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哺乳动物附植前胚胎的基因表达调控 总被引：4，自引：0，他引：4
哺乳动物胚胎附植前期包括:合子的形成,胚胎基因组的激活和细胞分化的开始。在这个时期,发育由母源物质控制转为合子基因控制,在此过程,同时形成染色质介导的转录抑制时期,要解除抑制必须经过胚胎基因组的激活。通过对体内、外附植前胚胎的mRNA的表达特点以及它们与成功发育联系的研究,可以筛选出最佳的体外培养条件,设计最佳的核移植方案。 相似文献
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胚胎分割是将哺乳动物附植前胚胎分成若干个具有继续发育潜力部分的生物技术,也是胚胎移植常用的辅助技术。运用胚胎分割可增加有限的优质胚胎数目以获得同卵孪生后代〔1〕。运用同卵孪生后代作实验材料,可消除遗传差异,提高实验结果的准确性,更为重要的是通过胚胎的显微切割可 相似文献
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用８～１２周龄ＢＡＬＢ／ｃ公鼠采取皮肤移植和脾细胞腹腔注射２种方式免疫同日龄纯系ＢＡＬＢ／ｃ母鼠,制备Ｈ－Ｙ抗血清。经精子微量细胞毒性试验,筛选的抗血清用于胚胎体外培养,以鉴定附植前小鼠胚胎上Ｈ－Ｙ抗原的表达。选用８ｃｅｌｌ－早期囊胚进行体外培养,分别于５～６、１８和２４ｈ观察胚胎发育情况。５～６ｈ后,Ｈ－Ｙ抗血清培养的胚胎退化率高达４０％,且发育迟缓,与自然性比例差异不明显（Ｐ〉０．０５）。正常 相似文献
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Mihiro SHIBUTANI Jibak LEE Takashi MIYANO Masashi MIYAKE 《The Journal of reproduction and development》2015,61(2):106-115
The embryo culture technique has been improving, but the detailed demands for energy substrates such as glucose, fructose, pyruvate and lactate of preimplantation embryos are still unclear. In the present study, the demands of pig preimplantation embryos at each different developmental stage were investigated by use of parthenogenetic diploids as a model of pig preimplantation embryos. Pig parthenogenetic diploids showed different use of glucose and fructose before and after the 4-cell stage. Although glucose supported the development of pig embryos throughout the preimplantation stages and even maintained the expansion and hatching of blastocysts, it suppressed development to the blastocyst stage when glucose coexisted with pyruvate and lactate from 4 h after activation, but not after 48 h (early 4-cell stage). Since ketohexokinase that metabolizes fructose was not expressed in 2-cell and 4-cell diploids, a medium that included only fructose as a major
energy substrate did not support early cleavage of pig diploids beyond the 4-cell stage, and almost no diploids developed to the morula stage just as in a medium without carbohydrates. These results may explain the different suppressive effects on pig preimplantation development between glucose and fructose when pyruvate and lactate were present in a medium. In addition, 4-cell diploids that had been cultured in a medium with pyruvate and lactate developed to the expanded blastocyst stage without any carbohydrates as a major energy substrate. These results show that the demands for carbohydrates are different depending on the developmental stage in pig preimplantation embryos. 相似文献
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Omar Khadrawy Samuel Gebremedhn Dessie Salilew-Wondim Franca Rings Christiane Neuhoff Michael Hoelker Karl Schellander Dawit Tesfaye 《Reproduction in domestic animals》2020,55(10):1275-1285
Nrf2 is a master regulator for antioxidant machinery against oxidative stress in bovine preimplantation embryos. The endogenous or exogenous modulation of Nrf2-KEAP1 system in bovine embryos may contribute to the understanding of the mechanisms behind the response of embryos to stress conditions. Therefore, here we aimed to investigate the protective effect of quercetin on bovine preimplantation embryos exposed to higher atmospheric oxygen concentration. For that, blastocysts, which were developed from zygotes cultured in media supplemented with or without quercetin under high oxygen level (20%), were subjected intracellular ROS level and mitochondrial analysis, and determining blastocyst formation rate and total cell number. Moreover, mRNA and protein expression level of Nrf2 and selected downstream antioxidant genes were investigated in the resulting blastocysts. Quercetin supplementation in vitro culture did not affect cleavage and blastocyst rate until day 7. However, quercetin supplementation resulted in higher blastocyst total cell number and reduction of intracellular ROS level accompanied by increasing mitochondrial activity compared with control group in both day 7 and day 8 blastocysts. Moreover, quercetin supplementation induced mRNA and protein of Nrf2 with subsequent increase in the expression of downstream antioxidants namely: NQO1, PRDX1, CAT and SOD1 antioxidants. In conclusion, quercetin protects preimplantation embryos against oxidative stress and improves embryo viability through modulation of the Nrf2 signalling pathway. 相似文献
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家畜胚胎性别鉴定的研究进展 总被引：5，自引：0，他引：5
家畜早期胚胎的性别鉴定是家畜胚胎移植的重要内容 ,对实现动物性别的人为控制具有重要意义。近年来随着性别鉴定技术的不断发展 ,出现了许多鉴定方法。本文对性别决定基因、动物性别决定机理和早期胚胎性别鉴定的方法作一简要综述 ,特别是对PCR方法在胚胎性别鉴定中的应用及其基本过程作一简述 相似文献
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Mammalian preimplantation embryos enter the S phase immediately after the end of the M phase; their cell cycle lacks a substantial G1 phase. Previously, we suggested that the absence of the G1 phase was attributable to a loss of retinoblastoma protein (RB) function, which is required for suppression of S phase entrance and that this loss of RB function in turn was attributable to the low RB expression level during preimplantation development in mouse embryos. The present study aimed to examine whether or not RB inhibition by CDK4/6-cyclin D-dependent phosphorylation is involved in the loss of RB function in preimplantation mouse embryos by the expression of p16(INK4a), a potent endogenous inhibitor of CDK4/6-cyclin D. First, the decrease in RB expression between the four-cell and morula stages was confirmed in in vivo-derived mouse embryos. We then examined the efficiency of the p16(INK4a) expression vector in inhibiting RB phosphorylation and cell cycle progression using NIH-3T3 cells and obtained gradual RB dephosphorylation and a significantly lower proliferation rate in p16(INK4a)-transfected cells than in control cells. This indicated the successful p16(INK4a) effects on cell-cycle progression by the vector used. On the other hand, the development rate of mouse embryos injected with the p16(INK4a) expression vector was the same as that of the control embryos, although p16(INK4a) expression was detected at mRNA and protein levels in the former group but not in the control group. These results suggest that RB phosphorylation is not involved in RB dysfunction or in the lack of a G1 phase in mouse embryos and that the decrease in RB expression is important for preimplantation-embryo-specific cell-cycle regulation. Moreover, the present study indicates the similarity between preimplantation embryos and cancer cells, which p16(INK4a) expression does not arrest at the G1 phase. 相似文献
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Tom Seeling Elisa Haucke Alexander Navarrete Santos Katarzyna J. Grybel Jacqueline Gürke S. Mareike Pendzialek Maria Schindler Andreas Simm Anne Navarrete Santos 《Reproduction in domestic animals》2019,54(Z3):4-11
In a diabetic pregnancy, an altered maternal metabolism led to increased formation of reactive α‐dicarbonyls such as glyoxal (GO) and methylglyoxal (MGO) in the reproductive organs and embryos. The enzyme glyoxalase (GLO) 1 detoxifies reactive α‐dicarbonyls thus protecting cells against malfunction or modifications of proteins by advanced glycated end products (AGEs). The aim of this study was to analyse the influence of a maternal insulin‐dependent diabetes mellitus (IDD) on GLO1 expression and activity in preimplantation embryos in vivo and human trophoblast cells (Ac‐1M88) in vitro. Maternal diabetes was induced in female rabbits by alloxan before conception and maintained during the preimplantation period. GLO1 expression and activity were investigated in 6‐day‐old blastocysts from healthy and diabetic rabbits. Furthermore, blastocysts and human trophoblast cells were exposed in vitro to hyperglycaemia, GO and MGO and analysed for GLO1 expression and activity. During gastrulation, GLO1 was expressed in all compartments of the rabbit blastocyst. Maternal diabetes decreased embryonic GLO1 protein amount by approx. 30 per cent whereas the enzymatic activity remained unchanged, indicating that the specific GLO1 activity increases along with metabolic changes. In in vitro cultured embryos, neither hyperglycaemia nor MGO and GO had an effect on GLO1 protein amount. In human trophoblast cells, a stimulating effect on the GLO1 expression was shown in the highest GO concentration, only. Our data show that maternal diabetes mellitus affects the specific activity of GLO1, indicating that GLO1 was post‐translationally modified due to changes in metabolic processes in the preimplantation embryos. 相似文献
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Epigenetic characteristics of cloned and in vitro-fertilized swamp buffalo (Bubalus bubalis) embryos
Suteevun T Parnpai R Smith SL Chang CC Muenthaisong S Tian XC 《Journal of animal science》2006,84(8):2065-2071
Swamp buffalos are becoming endangered due to reproductive inefficiencies. This is of concern because many countries depend heavily on their products. Somatic cell nuclear transfer (SCNT) is a potential strategy for preserving endangered species. To date, SCNT in swamp buffalo has succeeded in the creation of blastocyst embryos. However, development to term of SCNT swamp buffalos is extremely limited, and only 1 live birth has been reported. An abnormal epigenetic mechanism is suspected to be the cause of developmental failure, as is also seen in other species. The DNA methylation and histone acetylation are key players in epigenetic modification and display marked variability during embryonic preimplantation development. Knowledge of epigenetic modifications will aid in solving the developmental problems of SCNT embryos and improving reproductive technology in the swamp buffalo. The objective of this study was to determine the relationship between preimplantation embryonic development and 2 epigenetic patterns, global DNA methylation and histone acetylation, in SCNT and in vitro-fertilized (IVF) swamp buffalo embryos. In addition, we examined the correlations between those 2 mechanisms in the SCNT and IVF swamp buffalo embryos throughout the developmental stages using double immunostaining and quantification of the emission intensities using confocal microscopy. We discovered an aberrant methylation pattern in early preimplantation-stage swamp buffalo SCNT embryos. In addition, greater variability in the DNA methylation levels among nuclei within SCNT embryos was discovered. Hyperacetylation was also observed in SCNT embryos compared with IVF embryos at the 4- and 8-cell stages (P < 0.05). Dynamic changes and interplay between these 2 epigenetic mechanisms could be crucial for embryonic development during the early preimplantation period. The aberrancies uncovered here may contribute to the low efficiency of SCNT. 相似文献