Inhibition of mouse blastocyst attachment and outgrowth by protease inhibitors

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Abstract
Effects of protease inhibitors on development of mouse blastocysts and fibrinolytic activity of trophoblast were examined by growing embryos on monolayers of decidual cells in the presence of inhibitors. Nitrophenol-p-guanidino benzoate (NPGB) was the most effective inhibitor; 10−4 M NPGB inhibited attachment and trophoblast outgrowth by 24% and 66%, respectively, and inhibited the fibrinolytic activity of trophoblast by 86%. The effects of NPGB were reversible, as demonstrated by the embryos' ability to attach and resume normal development when transferred to NPGB-free medium. Soybean trypsin inhibitor and ϵ-aminocaproic acid were less effective than NPGB in inhibiting blastocyst development. When 10−4 M NPGB and 350 μg/ml of soybean trypsin inhibitor were added together, blastocyst development and fibrinolytic activity were inhibited more severely than when either inhibitor was added alone. We suggest that at least two types of proteolytic activities in mouse blastocysts are involved in implantation, attachment requiring trypsin-like activity, and trophoblast outgrowth requiring both plasminogen activator and trypsin-like activity.