Effects of tumour necrosis factor‐alpha and interleukin‐1 beta on in vitro development of bovine secondary follicles

Abstract

The objective of this study was to determine the effects of TNF-α and IL-1β on development and survival of bovine secondary follicle culture in vitro for 18 days. Secondary follicles (~0.2 mm) were isolated from ovarian cortex and individually cultured at 38.5°C, with 5% CO in air, for 18 days, in TCM-199 alone (cultured control) or supplemented with 10 ng/ml IL-1β, 10 ng/ml TNF-α or both TNF-α and IL-1β. The effects of these treatments on growth, follicular survival, antrum formation, viability, ultrastructure and mRNA levels for GDF-9, c-MOS, H1foo and Cyclin B1 were evaluated. The results showed that addition of TNF-α to culture medium increased follicular diameter and rate of antrum formation, whereas that of IL-1β and a mixture of IL-1β and TNF-α did not do so. Ultrastructural analysis showed that, among the tested cytokine treatments, follicles cultured in the presence of TNF-α had the best-preserved oocytes and granulosa cells. The presence of TNF-α, IL-1β or both did not influence the expression of mRNAs analysed. In conclusion, in contrast to IL-1β, TNF-α promotes growth of and antrum formation in in vitro cultured bovine secondary follicles, while their ultrastructure and viability were maintained.

DNA methylome profiling of granulosa cells reveals altered methylation in genes regulating vital ovarian functions in polycystic ovary syndrome

“…However, these studies utilized TNF at 10–20-fold higher doses relative to its physiological levels. In alternate studies, optimal TNF levels have been reported to impart a protective function in the maintenance of bovine GCs and oocytes [64], facilitate ovulation [65], and increase the GC proliferation in animal models [66]. Decreased levels of endogenous TNF in GCs has been attributed to diminished oocyte competence due to a reduction in its downstream effector, i.e.…”

Interleukin-1β and TNF-α systems in ovarian follicles and their roles during follicular development, oocyte maturation and ovulation

“…Previous studies have revealed that IL-1β controls the transition of follicular cells by facilitating their proliferation and differentiation (Brännström, 2004). For bovine secondary follicles, Paulino et al (2018) showed that IL-1β does not influence their development in vitro. Probably, IL-1β acts mainly in large antral follicles.…”

“…In mice, Nilsson et al (2006) demonstrated that TNF-α can interact with P4 to regulate the growth of primordial follicles. In bovine secondary follicles in vitro, Paulino et al (2018) showed that TNF-α promotes growth, antrum formation and maintains the ultrastructure of secondary follicles in vitro. These studies show that the effect of TNF-α on ovarian cells is dependent on the stage of follicle development, which can be associated with different levels of TNF-α receptor expression (Witty et al, 1996).…”

Effects of dexamethasone on growth, viability and ultrastructure of bovine secondary follicles cultured in vitro

“…These ovaries were collected immediately after the death of the cows and were washed in 70% ethanol for about 10 s and twice in sterile 0.9% saline solution. Subsequently, the ovaries were transported within 1 h to the laboratory in tubes containing 0.9% saline solution supplemented with penicillin (100 mg/ml) and streptomycin (100 mg/ml) at 4°C (Paulino et al, 2018). This study was carried out in accordance with procedures approved by the Ethics and Animal Welfare Committee of the Federal University of Ceara.…”