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Submitted on February 21, 2007
Accepted on April 23, 2007
Obstetrics and Gynaecology, Department of Reproductive and Developmental Sciences, University of Edinburgh (M.M., E.G., W.C.D.) and Medical Research Council Human Reproductive Sciences Unit (A.S.M., H.M.F.), Centre for Reproductive Biology, Queen's Institute of Medical Research, Little France, Edinburgh EH16 4TJ, United Kingdom
* To whom correspondence should be addressed. E-mail: m.myers{at}sms.ed.ac.uk.
Luteolysis in women is associated with an up-regulation of the expression and activity of MMP-2 which is inhibited by hCG during maternal recognition of pregnancy. As the primary source of MMP-2 is fibroblasts that do not express LH/hCG receptors, we aimed to investigate the regulation of MMP-2. Women with regular cycles having hysterectomy for non-malignant conditions and women undergoing oocyte retrieval for assisted conception were used in this current study. Novel primary cultures and co-cultures of luteinized granulosa cells and fibroblast-like cells in conjunction with human corpora lutea from different stages of the luteal phase were used to investigate the role of activin-A in the corpus luteum. The effect of hCG, activin-A and follistatin on MMP-2 activity and expression was assessed by gelatin zymography and quantitative RT-PCR in primary cell cultures. Confirmation of signaling pathways involved in the activation of MMP-2 were assessed by immunofluorescence, RT-PCR and quantitative RT-PCR. In primary cell culture, steroidogenic cells secrete activin-A and its inhibitors, inhibin-A and follistatin. Follistatin expression is up-regulated by hCG (P < 0.05). The fibroblast-like cells producing MMP-2 have the machinery for activin reception, expressing both type I and type II activin receptors and Smad proteins. Activin-A up-regulated both activity and expression of MMP-2 in fibroblast-like cells (P < 0.05). This activity was inhibited in co-cultures of luteinized granulosa cells and fibroblast-like cells in the presence of hCG (P < 0.05) or follistatin (P < 0.01). Activin-A is an excellent candidate for an effector molecule in human luteolysis whose paracrine action is inhibited during maternal recognition of pregnancy.
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