Long Noncoding RNA LncPGCR mediated by TCF7L2 regulates chicken PGCs formation

Abstract

Background: Several of the thousands of long noncoding RNAs (lncRNAs) have been functionally characterized, yet its specific function and molecular mechanism in the formation of chicken primordial germ cells (PGCs) remains poorly understood. In the present study, we aim to investigate the role of LncPGCR (LncRNA PGCs Regulator) in PGCs formation.Methods: LncPGCR, Cvh, Nanog, C-kit, gga-mir-6577-5p and Btrc expressions were detected by qRT-PCR. The percentage of PGCs cells was detected by flow cytometry, immunocytochemistry, and PAS staining. The interaction of histone acetylation, DNA methylation, transcription factor TCF7L2, and LncPGCR was confirmed by Luciferase reporter assay. The interaction of GAPDH and LncPGCR was measured by RNA pull-down, RIP, and Western blot.Results: We observed the increased expression of LncPGCR in PGCs. It is mainly expressed in the cytoplasm and encodes small peptides. Moreover, over-expression of LncPGCR could promote PGCs formation in vitro and in vivo. Besides, we first reported that histone acetylation, DNA methylation, and transcription factor TCF7L2 can regulate LncPGCR expression in PGCs. In addition, LncPGCR activates WNT signaling pathways to promote PGCs formation by adsorbing gga-mir-6577-5p, relieving its inhibitory effect on target gene Btrc. Meanwhile, LncPGCR contributed to PGCs formation by increasing the phosphorylation level of GAPDH to activate the TGF-β signal pathway.Conclusion: LncPGCR over-expression promoted ESCs differentiation into PGCs through the potential LncPGCR/miR-6577-5p/BTRC pathway or increasing the phosphorylation level of GAPDH to activate the TGF-β signal pathway.