Pluripotency and embryonic lineage genes expression in the presence of small molecule inhibitors of FGF, TGFβ and GSK3 during pre-implantation development of goat embryos

• Published in: Gene Expression Patterns 2023-12, Vol.50, p.119334-119334, Article 119334
• Main Authors: Hajian, Mehdi, Rouhollahi Varnosfaderani, Shiva, Jafarpour, Farnoosh, Tanhaei Vash, Nima, Nasr-Esfahani, Mohammad Hossein
• Format: Article
• Language: English
• Subjects: Animals; blastocyst; Blastocyst - metabolism; cellular agriculture; embryogenesis; Embryonic Development - genetics; Embryonic stem cells; FGF; GATA transcription factors; gene expression; genome; Glycogen Synthase Kinase 3 - metabolism; Glycogen Synthase Kinase 3 - pharmacology; Goat blastocyst; goats; Goats - genetics; GSK3B; medicine; species; TGFβ; Transforming Growth Factor beta - metabolism; Transforming Growth Factor beta - pharmacology
• ISSN: 1567-133X; 1872-7298
• DOI: 10.1016/j.gep.2023.119334

Generating stable livestock pluripotent stem cells (PSCs) can be used for complex genome editing, cellular agriculture, gamete generation, regenerative medicine and in vitro breeding schemes. Over the past decade, significant progress has been made in characterizing pluripotency markers for livestock species. In this study, we investigated embryo development and gene expression of the core pluripotency triad (OCT4, NANOG, SOX2) and cell lineage commitment markers (REX1, CDX2, GATA4) in the presence of three small molecules and their combination [PD0325901 (FGF inhibitor), SB431542 (TGFβ inhibitor), and CHIR99021 (GSK3B inhibitor)] from day 2–7 post-insemination in goat. Significant reduction in rate of blastocyst formation was observed when SB was used along with PD or CHIR and their three combinations had more sever effect. SB and CHIR decreased the expression of SOX2 while increasing the GATA4 expression. PD decrease the relative expression of NANOG, OCT4 and GATA4, while increased the expression of REX1. Among the combination of two molecules, only SB + CHIR combination significantly decreased the expression of GATA4, while the combination of the three molecules significantly decreases the expression of NANOG, SOX2 and CDX2. According to these results, the inhibition of the FGF signaling pathway, by PD may lead to blocking the hypoblast formation as observed by reduction of GATA4. OCT4 and NANOG expressions did not show signs of maintenance pluripotency. GATA4, NANOG and OCT4 in the PD group were downregulated and REX1 as EPI-marker was upregulated thus REX1 may be considered as a marker of EPI/ICM in goat. •In goat, we particularly showed that inhibition of TGFβ signaling with SB431542 along with PD0325901 and CHIR99021 lead to significant decrease in blastocyst formation.•SB431542 decreased the expression of SOX2 and this effect could have been mediated through inhibition of trophoblast formation.•The combination of the three molecules (SB431542, PD0325901, and CHIR99021) significantly decreases the expression of NANOG, SOX2 and CDX2.