Levels of mRNA for bone morphogenetic proteins, their receptors and SMADs in goat ovarian follicles grown in vivo and in vitro

Abstract

This study investigated the stability of housekeeping genes (glyceraldehyde-3-phosphate dehydrogenase, β-tubulin, β-actin, phosphoglycerate kinase (PGK), 18S rRNA, ubiquitin and ribosomal protein 19) and the levels of mRNA for bone morphogenetic protein-2 (BMP-2), -4 (BMP-4), -6 (BMP-6), -7 (BMP-7) and -15 (BMP-15), their receptors (BMPR-IA, -IB and -II) and Similar to Mothers Against Decapentaplegic (SMADs) (-1, -5 and -8) in goat follicles of 0.2, 0.5 and 1.0mm, as well as in secondary follicles before and after culture for 18 days. β-tubulin and PGK were the most stable housekeeping genes and the levels of mRNA for BMP-2 in follicles of 0.2mm were higher than in follicles of 0.5 and 1.0mm. For BMP-4, -6 and -7, the highest levels of mRNA were found in follicles of 1.0mm. The expression of BMPR-IB was higher in follicles of 0.2mm, whereas the levels of BMPR-II were higher in follicles of 0.5mm. The levels of mRNA for SMAD-5 were higher in follicles of 0.2mm, whereas SMAD-8 had higher levels in 0.5-mm follicles. After culture, follicles showed increased levels of mRNA for BMP-2 and reduced mRNA for BMP-4, BMP-7, BMPR-IA and SMAD-5. In conclusion, β-tubulin and PGK are the most stable reference genes, and BMPs, their receptors and SMADs have variable levels of mRNA in the follicular size classes analysed.

Bone Morphogenetic Protein‐6 (BMP‐6) Stimulates the Antrum Formation by the Regulation of its Signalling Pathway in Caprine Pre‐antral Follicles Cultured In Vitro

“…() and Costa et al. () observed similar results, showing the down‐regulation of BMPR1A and BMPR2 expression after in vitro culture of ovine granulosa cells and caprine secondary follicles, respectively. Previous studies demonstrated that BMP‐6 plays a role in follicular development through the proliferation and differentiation of granulosa cells (Juengel et al.…”

“…The results of a recent study showed that the mRNA expression for BMP6 gene increases with in vivo follicular growth from primordial to secondary follicle stage (Costa et al. ), indicating the importance of BMP‐6 during the follicular growth in goats. Moreover, the protein and mRNA expression of BMP6 was demonstrated in the caprine oocytes of follicles during all stages of development, particularly in those from antral follicles (Frota et al.…”

“…The caprine pre‐antral and antral follicles expressed mRNA for SMAD1 , SMAD5 and SMAD8 in vivo and in vitro after culture using only rFSH added to the follicular culture medium (Costa et al. ). In cattle, antral follicles expressed SMAD1 protein, and its expression was activated through BMP‐6 in granulosa cells (Glister et al.…”

“…Recently, Frota et al (2013) showed that the addition of rFSH to the culture medium increased the mRNA expression for BMP6 gene in isolated caprine pre-antral follicles. In other study, Costa et al (2012) studied the mRNA expression for SMAD1, SMAD5 and SMAD8; BMP2, BMP4, BMP6, BMP7 and BMP15; and BMPR1A, BMPR1B and BMPR2 genes in a basic culture medium containing only rFSH. However, to our knowledge, there are no reports in goats regarding the effect of BMP-6 of its own on the mRNA expression for the whole BMP receptors/Smad signalling pathway, which includes BMPR1A, BMPR2, SMAD1, SMAD4, SMAD5, SMAD6, SMAD7 and SMAD8, in the in vitro culture of caprine-isolated secondary follicles.…”

Felis catus ovary as a model to study follicle biology in vitro

“…Follicle culture at different stages of development has been used to study diverse biological processes (folliculogenesis, steroidogenesis, and oocyte maturation) in various species, such as goat [9,10], mouse [11][12][13], rat [14], sheep [15,16], dog [17][18][19], and monkey [20][21][22][23]. Regarding the domestic cat, efforts to date have focused on oocytes [7,[24][25][26][27][28][29][30] or preantral follicle cultures [19,31].…”