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Biology of Reproduction, Vol 50, 88-94, Copyright © 1994 by Society for the Study of Reproduction
ARTICLES |
M Brannstrom, RJ Norman, RF Seamark and SA Robertson
Department of Obstetrics and Gynaecology, University of Adelaide, Queen Elizabeth Hospital, Woodville, Australia.
To examine the production of cytokines by the ovary during ovulation, ovaries were obtained from immature rats and from eCG/hCG-primed immature rats at different stages of the ovulatory process (before hCG injection, 10 h after hCG, and 20 h after hCG) and were perfused in vitro for 5 h. Large quantities of interleukin (IL)-6 and granulocyte- macrophage colony-stimulating factor (GM-CSF) bioactivity and smaller amounts of tumor necrosis factor-alpha (TNF alpha) and IL-1 bioactivity were found in the perfusate. IL-2 and IL-3 were not detectable in the perfusion media. The GM-CSF content was significantly higher in the perfusate of ovulating ovaries (obtained 10 h after hCG) compared to the earlier stages. Studies on preovulatory ovaries (prior to hCG injection) revealed that GM-CSF release was not influenced by LH, but was markedly increased when recombinant human IL-1 beta (4 ng/ml) was added to the perfusion medium. IL-6 was released in similar amounts from ovaries at all stages. The identity of bioactive GM-CSF was confirmed by neutralization with a specific polyclonal antibody against murine GM-CSF. Size-exclusion chromatography of perfusion medium revealed peaks of GM-CSF and IL-6 bioactivity at approximate molecular masses of 21-23 kDa and 24-25 kDa, respectively. This study demonstrates that the rat ovary produces IL-6, GM-CSF, TNF alpha, and IL-1 prior to and during the ovulatory process and that there are temporal fluctuations in GM-CSF release with a peak in output at ovulation.
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