Manganese inhibits spontaneous nuclear maturation in bovine cumulus-enclosed oocytes

Abstract

The objective of this work was to examine the effects of manganese concentration on nuclear maturation of bovine cumulus-enclosed (CEO) and denuded oocytes cultured for 7 h or 22 h. Following culture, oocytes were then fixed and stained for assessment of nuclear maturation stage. The addition of MnCl2 significantly suppressed nuclear maturation after 7h of culture (15, 69, 84 and 70% of oocytes were still at the germinal vesicle stage after culture with 0, 50 μM, 0.5 mM and 5mM MnCl2, respectively; P < 0.001). However, MnCl2 was without significant effect on denuded oocytes cultured for 7 h. When CEO and denuded oocytes were cultured with manganese for 22 h, the percentages of mature oocytes were reduced (96 , 20, 15, 3% and 80, 39, 53 and 16% for CEO and denuded oocytes cultured with 0, 50 μM, 0.5 mM and 5 mM MnCl2, respectively; P < 0.0005). The inhibitory effect of 50 μM MnCl2 was transient and reversible because it did not maintain oocytes in meiotic arrest after 22 h of culture. In addition, 72% of the CEO cultured with 50 μM MnCl2 for 7 h and subsequently cultured without manganese for 18 h were mature. The concentration of manganese (6 ± 1 μM) in follicular fluid (as determined by atomic absorption spectrophotometry) was below inhibitory concentrations. In conclusion, manganese inhibited germinal vesicle breakdown in bovine CEO; however, only the effect of the lowest concentration tested (50 μM) was reversible. Key words: Bovine, oocyte, meiosis, manganese, follicular fluid