Dynamics of the transcriptome during chicken embryo development based on primordial germ cells
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(2020) 13:441 Dunislawska et al. BMC Res Notes https://doi.org/10.1186/s13104-020-05286-w
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RESEARCH NOTE
Dynamics of the transcriptome during chicken embryo development based on primordial germ cells Aleksandra Dunislawska, Agata Szczerba, Maria Siwek and Marek Bednarczyk*
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, Primordial germ cells, Microarray, White Leghorn Introduction Primordial germ cells (PGCs) are the earliest recognisable precursors of adult germ cells. They transfer the genetic information to the next generation of cells. Chicken PGCs originate from the epiblast [1]. They are located at the centre of the area pellucida at stage X of Hamburger-Hamilton (HH) [2] and are translocated anteriorly to the germinal crescent [3]. Subsequently, the PGCs localise in the vascular system and use those extraembryonic blood vessels as a vehicle to reach the germinal ridges (Additional file 1: Figure S1). They accumulate in germinal ridges as gonadal PGCs (gPGCs), are also termed as gonocytes [4] and differentiate into spermatogonia in males or oogonia in females. Circulating *Correspondence: [email protected] Department of Animal Biotechnology and Genetics, UTP University of Science and Technology, Bydgoszcz, Poland
PGCs and gPGCs are functionally very similar. As precursors of reproductive cells, PGCs are an important tool in the study on reproduction of vertebrates and in gene expression and epigenetic studies. The first challenge in gaining knowledge and understanding of the mechanisms associated with PGCs is to analyse the changes that may occur in a given location in the embryo on the basis of PGCs from a given place, regardless of gender. PGCs migrate into the bloodstream before they reach the gonads and are classified as circulating PGCs (cPGCs). However, the regulation of gene expression during th
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