Comparison of the MicroRNA Expression Profiles of Male and Female Avian Primordial Germ Cell Lines

Abstract

Primordial germ cells (PGCs) are the precursors of adult germ cells, and among the embryonic stem-like cells in the bird embryo, only they can transmit the genetic information to the next generation. Despite the wide range of applications, very little is known about the mechanism that governs primordial germ cell self-renewal and differentiation. As a first step, we compared 12 newly established chicken PGC lines derived from two different chicken breeds, performing CCK-8 proliferation assay. All of the lines were derived from individual embryos. A significant difference was found among the lines. As microRNAs have been proved to play a key role in the maintenance of pluripotency and the cell cycle regulation of stem cells, we continued with a complex miRNA analysis. We could discover miRNAs expressing differently in PGC lines with high proliferation rate, compared to PGC lines with low proliferation rate. We found that gga-miR-2127 expresses differently in female and male cell lines. The microarray analysis also revealed high expression level of the gga-miR-302b-3p strand (member of the miR-302/367 cluster) in slowly proliferating PGC lines compared to the gga-miR-302b-5p strand. We confirmed that the inhibition of miR-302b-5p significantly increases the doubling time of the examined PGC lines. In conclusion, we found that gga-miR-181-5p, gga-miR-2127, and members of the gga-miR-302/367 cluster have a dominant role in the regulation of avian primordial germ cell proliferation.

miR-302d Competitively Binding with the lncRNA-341 Targets TLE4 in the Process of SSC Generation

“…These studies have shown that miR-302 family has high specificity or differentiation expression patterns in mammalian ESCs, and it is consistent with our RNA-seq data in chicken which shows the differentially high expression of miR-302d in chicken ESCs. A small number of studies have also shown that gga-miR-181-5p, gga-miR-2127, and gga-miR-302/367 clusters play a leading role in the regulation of proliferation of avian primitive germ cells [ 25 ]. Both gga-miR-302b and gga-miR-17-5p can regulate glucose phosphate isomerase and affect the proliferation of PGCs [ 26 ] which suggests that miR-302d may play an important role in the differentiation of chicken male germ cells.…”

Comprehensive analysis of miRNAs, lncRNAs, and mRNAs reveals potential players of sexually dimorphic and left-right asymmetry in chicken gonad during gonadal differentiation

“…MiR-302 b and miR-17- 5p regulated Glucose Phosphate Isomerase (GPI) in chicken PGCs proliferation ( Rengaraj et al., 2013 ). MiR-181-5p, miR-2127, and members of the miR-302/367 cluster had a dominant role in the regulation of PGCs proliferation ( Lee et al., 2011 , Lazar et al., 2018 ). The number of PGCs and germ cells proliferated rapidly, and in the left gonad, it was greater than that in the right gonad during gonadal differentiation ( Vallisneri et al., 1990 ).…”

“…However, there was little known about the contributions of miRNAs and lncRNAs in left-right asymmetry in chicken. Several miRNAs were reported to play key role in testicular development and the proliferation of primordial germ cells, such as miR-202, miR-107 and miR-302b ( Bannister et al., 2011 , Miao et al., 2016 , Lazar et al., 2018 ). Currently, growing evidences indicated that lncRNAs might have contributed to ovarian development ( Bouckenheimer et al., 2018 , Li et al., 2018 , Liu et al., 2018 ).…”

Investigation of the Guinea fowl and domestic fowl hybrids as potential surrogate hosts for avian cryopreservation programmes

“…61 . A male GFP expressing cell line (4ZP) was selected for injection 62 . Hybrid eggs were incubated to stage HH15-16.…”