Department of Animal Biotechnology and Genetics, UTP University of Science and Technology, Bydgoszcz, Poland.
BMC Res Notes. 2020 Sep 18;13(1):441. doi: 10.1186/s13104-020-05286-w.
Regulation of gene expression during embryo development on the basis of migration of primordial germ cells (PGCs) in vivo has been rarely studied due to limited cell number and the necessity to isolate PGCs from a large number of embryos. Moreover, little is known about the comprehensive dynamics of the transcriptome in chicken PGCs during early developmental stages. The current study investigated transcriptome dynamics of chicken PGCs at key developmental stages: 4.5, 8 and 12 days of embryo incubation. PGCs were collected, and RNA was isolated using a commercial kit for single cells. The isolated RNA was subjected to microarray analysis (Agilent Technologies).
Between 8 and 12 days of incubation, the highest number of genes was regulated. These data indicate that the most intense biological activity occurs between 8 and 12 days of embryo development. Heat map showed a significant decrease in gene expression on day 8, while it increased on day 12. The development of a precise method to isolate bird PGCs as well as the method to isolate RNA from single cells isolated from one embryo allows for early molecular analysis and detection of transcriptome changes during embryonic development.
由于原始生殖细胞 (PGC) 的数量有限,且需要从大量胚胎中分离 PGC,因此,体内 PGC 迁移过程中的胚胎发育基因表达调控研究甚少。此外,人们对鸡 PGC 在早期发育阶段的转录组的综合动态知之甚少。本研究调查了鸡 PGC 在关键发育阶段的转录组动态:胚胎孵育的 4.5、8 和 12 天。使用商业试剂盒从单细胞中分离 RNA。分离的 RNA 进行微阵列分析(Agilent Technologies)。
在孵育的 8 至 12 天之间,调节了最多数量的基因。这些数据表明,胚胎发育的 8 至 12 天之间发生了最强烈的生物学活性。热图显示第 8 天的基因表达显着下降,而第 12 天则增加。开发一种精确的方法从单个胚胎中分离鸟类 PGC 以及从单个细胞中分离 RNA 的方法,可实现早期分子分析并检测胚胎发育过程中的转录组变化。