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Silva, A.W.B.; Ribeiro, R.P.; Menezes, V.G.; Barberino, R.S.; Passos, J.R.S.; Dau, A.M.P.; Costa, J.J.N.; Melo, L.R.F.; Bezerra, F.T.G.; Donato, M.A.M.; Peixoto, C.A.; Matos, M.H.T.; Gonçalves, P.B.D; van den Hurk, R.; Silva, J.R.V.
Animal reproduction science, July 2017, 2017-Jul, 2017-07-00, 20170701, Letnik: 182Journal Article
•TNF-α system members are expressed at different follicular stages in bovine ovaries.•TNF-α reduces follicular survival in cultured bovine ovarian tissue.•TNF-α increases the total number of apoptotic cells in cultured follicles.•Dexamethasone improves ultrastructure of bovine follicle cultured in vitro. This study was conducted to detect the protein expression of TNF-α system members (TNF-α/TNFR1/TNFR2) in bovine ovarian follicles and to evaluate the effects of TNF-α or dexamethasone on the survival and growth of primordial follicles in vitro, as well as on gene expression in cultured ovarian tissue. It was hypothesized that TNF-α induces follicular atresia in ovarian tissues cultured in vitro, and that dexamethasone suppresses the production of endogenous TNF-α, which can improve follicle viability in vitro. Ovarian fragments were cultured for 6days in α-MEM+ supplemented with TNF-α (0, 1, 10, 100 or 200ng/ml) or dexamethasone (0, 1, 10, 100 or 200ng/ml). After culture, the expression of mRNAs for BCL-2, BAX, P53, TNF-α, and CASP3 and CASP6 were evaluated. Immunohistochemical results showed that the TNF-α system members, were detected in bovine preantral and antral follicles. After 6days, the TNF-α (10ng/ml) treatment reduced the percentage of normal preantral follicles and increased the number of TUNEL-positive cells in cultured tissue. Dexamethasone (10ng/ml) during 6days of culture did maintain the percentage of normal follicles and the ultrastructure of follicles, while the presence of TNF-α or dexamethasone did not influence primordial follicle activation. However, TNF-α or dexamethasone had no effect on the levels of mRNA for P53, BCL-2, BAX and CASP6, in cultured tissues, but the presence of dexamethasone reduced the levels of CASP3 compared to ovarian slices cultured in control medium (α-MEM+). In conclusion, proteins of the TNF-α system are expressed at different bovine follicle stages. The addition of TNF-α in culture reduces follicle survival and increases the number of apoptotic cells in ovarian tissue, while the presence of dexamethasone maintains follicle ultrastructure in cultured tissue.
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JCR | SNIP | JCR | SNIP | JCR | SNIP | JCR | SNIP |
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