Recently, the application of chemical inhibitors against differentiation signaling pathways has improved mESC establishment. In this study, we applied TGF-β (SB431542) and BMP4 (Noggin) inhibitors from cleavage to blastocyst stage in bovine, goat, and sheep embryos. SB significantly decreased blastocyst rate and total cell number (TCN) in sheep blastocysts, while TCN alone decreased significantly in cattle blastocysts.
In contrast to SB, Noggin significantly improved cattle blastocyst development but decreased TCN. However, Noggin treatment led to a significant increase in NTC in sheep blastocysts. Regarding the pluripotency triad (OCT4, NANOG, SOX2) and cell lineage compromise (REX1, CDX2, GATA4), SB led to a significant reduction in SOX2 expression in goats and cattle, while Noggin increased at least one or two of the pluripotent markers in these species. . Taken together, these data suggest that inhibition of TGF-β by Noggin may be more favorable for stem cell derivation in farm animals.
To evaluate the mRNA expression of GDF9, BMP15, FGF2, and their main receptors, transforming growth factor receptor 1 (TGFβ-R1), bone morphogenetic protein receptor, type IB (BMPR-IB), and growth factor receptor 2 of fibroblasts (FGFR2) in bovine follicular cells.
Materials and methods.
Total RNA was isolated from pooled samples of oocytes (OO), cumulus cells (CC) from cumulus-oocyte complexes (COC), and follicular cell (PC) pellets from 70 ovaries obtained from 96 beef heifers, collected in a local slaughterhouse. The expression pattern of growth factors and their receptors in bovine follicular cells was evaluated by the reverse transcriptase-polymerase chain reaction (RT-PCR).
The mRNA transcripts encoding the GDF9, BMP15, FGF2, TGFβ-R1, BMPR-IB, and FGFR2 genes were detected by RT-PCR in all cells studied. This is the first time that the expression of TGFβ-R1 and BMPR-IB receptors has been reported in bovine oocytes.
The presence of growth factors and receptor transcripts in the cells studied indicates that these factors could act as paracrine and autocrine regulators of folliculogenesis.
Oocyte; granulosa cells; growth factors; PCR; bovine (Source: AGROVOC)