Dynamics of the transcriptome during chicken embryo development based on primordial germ cells

Abstract

Objective: 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).

Results: 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.

Keywords: Gene expression; Microarray; Primordial germ cells; White Leghorn.

Fig. 1

Individual values (heat maps) of genes expressed on day 4.5, 8 and 12

Fig. 2

Analysis of the relationship between proteins encoded by genes whose expression was upregulated in PGCs on day 8 of embryo development. Lines of interactions according to STING software: light blue–from curated databases; pink–experimentally determined; dark green–gene neighborhood; red–gene fusions; dark blue–gene co-occurrence; light green–textmining; black–co-expression; violett–protein homology