Time- and Dose-Dependent Effects of 17 Beta-Estradiol on Short-Term, Real-Time Proliferation and Gene Expression in Porcine Granulosa Cells

Abstract

The key mechanisms responsible for achievement of full reproductive and developmental capability in mammals are the differentiation and transformation of granulosa cells (GCs) during folliculogenesis, oogenesis, and oocyte maturation. Although the role of 17 beta-estradiol (E2) in ovarian activity is widely known, its effect on proliferative capacity, gap junction connection (GJC) formation, and GCs-luteal cells transformation requires further research. Therefore, the goal of this study was to assess the real-time proliferative activity of porcine GCs in vitro in relation to connexin (Cx), luteinizing hormone receptor (LHR), follicle stimulating hormone receptor (FSHR), and aromatase (CYP19A1) expression during short-term (168 h) primary culture. The cultured GCs were exposed to acute (at 96 h of culture) and/or prolonged (between 0 and 168 h of culture) administration of 1.8 and 3.6 μM E2. The relative abundance of Cx36, Cx37, Cx40, Cx43, LHR, FSHR, and CYP19A1 mRNA was measured. We conclude that the proliferation capability of GCs in vitro is substantially associated with expression of Cxs, LHR, FSHR, and CYP19A1. Furthermore, the GC-luteal cell transformation in vitro may be significantly accompanied by the proliferative activity of GCs in pigs.

The blood vessels development, morphogenesis and blood circulation are three ontologic groups highly up-regulated in porcine oocytes before in vitro maturation

“…Moreover it was defined that proper regulation of these processes is required for complete maturation of oocytes. Our previous studies have shown various aspects of changes occurring in oocytes during maturation [1][2][3] and impact of environmental changes on these processes [4][5]. Morphological and biochemical alterations associated with the ability of embryo to formation of blood vessels networks seem to play pivotal role in successful implantation, decidualization and early embryo -maternal interaction.…”

Molecular regulation of miR-378 on the development of mouse follicle and the maturation of oocyte in vivo

“…The relationship between generative hormones level and connexin (Cx) has been well studied. Estrogens could lead to the increased expression of Cx43 in rat [32][33][34] and pig [35]. The appearance of the gap and the decrease of the molecules expression level involved in interaction between granulosa cells and oocyte may have blocked the signaling, nutrition, growth factor and steroids transmitting between granulosa cells and oocytes, which induced the apoptosis of granulosa cells and affected the maturation of oocyte and resulted in reduced ovary size (Figure 2(a,b)), as well as oocyte maturation rate ( Figure 6(a,b)).…”

Analysis of fructose and mannose – regulatory peptides signaling pathway in porcine epithelial oviductal cells (OECs) primary cultured long-term in vitro

“…Indeed, we introduced the primary culture systems based on: (1) human ovarian granulosa cells, (2) porcine ovarian granulosa cells, (3) porcine endometrial and myometrial cells, and (4) porcine epithelial oviductal cells (OECs) [1][2][3][4][5].…”