构建不同脂肪含量的猪胚胎-一种手工模型
Reconstruction of pig embryos with different lipids - a handmade model
猪的卵母细胞中含有大量的中性脂滴,这些脂滴在卵母细胞成熟及早期胚胎发育过程中发挥重要的作用。本研究尝试构建不同脂滴含量的猪胚胎,为更好的研究脂滴的作用提供研究模型。结合手工克隆与离心去脂方法,获得了3种不同脂滴含量的重组胚胎:全脂胚胎(W-L)、半脂胚胎(H-L)、去脂胚胎(R-L)。化学激活后,3种胚胎分别在WOWs系统中培养6天,观察发育能力。结果显示,三种重组胚胎都具有发育到囊胚的能力。接下来,尝试以半脂胚胎作为模型,用以探究脂滴和线粒体在早期胚胎发育过程中的作用。首先,正常去核的半卵用MitoTracker Red CMXRos 染色,然后,用染色后正常半卵和未染色的去脂半卵以及体细胞三者构建半脂胚胎并培养。在培养过程中,依次取样并观察1细胞、2细胞、4细胞和囊胚四个阶段胚胎中脂滴和线粒体的分布情况。结果显示:在1细胞阶段,脂滴和线粒体都集中分布在一端细胞质中;2细胞阶段,72%的2细胞中线粒体局部分布,28%的2细胞中线粒体已均匀分布在细胞质中,然而此阶段的脂滴并未均匀分布;在4细胞和囊胚中,脂滴和线粒体都趋向于均匀分布在整个细胞质中。因此, 构建不同脂滴含量的胚胎是适宜的脂滴研究模型,可以借助这种模型来研究脂滴、线粒体、内质网以及各细胞器之间的相互作用。?????
Numerous neutral lipids were stored in cytoplasm of porcine oocytes in the form of lipid droplets (LDs). These LDs play an important role in vitro maturation and embryo development. Reconstructed embryos with different lipids were performed in present study to establish a model for study of LDs. Three groups of reconstructed embryos were obtained by hand-made cloning (HMC) and delipation: W-L: whole LDs in embryos; H-L: half LDs in embryos; R-L: removed LDs in embryos. After chemical activation (0h), reconstructed embryos were cultured in WOWs system for 6 days. The in vitro developmental competence of the three kinds of reconstructed was assessed. Results showed that: each kind of these three groups of embryos can develop to blastocyst stage. Therefore, the embryos with other content of LDs can be reconstructed in future studies. Then, the H-L group embryos, which as a model to study the roles of LDs and mitochondria in early development of embryos, were composed of three parts: a half normal oocyte (stained by MitoTracker Red CMXRos before bisecting) without nucleus, a half of delipated oocyte without nucleus and a donor cell. Following, 1-cell (12h), 2-cell (24h), 4-cell (36h) and blastocyst (144h) were collected to observe the distribution of LDs and mitochondria. The results showed that LDs and mitochondria were successfully tracked from 1-cell to blastocyst stage. Both LDs and mitochondria were only visible in normal cytoplasm at 1-cell stage. At 2-cell stage, mitochondria were uniformly distributed in the cytoplasm in 28% of the total number. But the LDs still not evenly distributed. However, both LDs and mitochondria were dispersed fully at 4-cell and blastocyst stage. In conclusion, reconstructed embryos with different lipids were appropriate models for lipids study, as well as investigate the interaction among lipids, mitochondria and other organelles. ?????
曾亚琼、汪成飞、迟大明、牛英杰、李娟
畜牧、动物医学细胞生物学生物科学研究方法、生物科学研究技术
繁殖脂滴重组胚胎线粒体?????
lipid dropletsreconstructed embryosmitochondria?????
曾亚琼,汪成飞,迟大明,牛英杰,李娟.构建不同脂肪含量的猪胚胎-一种手工模型[EB/OL].(2016-03-30)[2025-08-23].http://www.paper.edu.cn/releasepaper/content/201603-438.点此复制
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