The expression of the placental anticoagulant protein, annexin V, by villous trophoblasts: immunolocalization and in vitro regulation.

We evaluated the histological and ultrastructural localization of the potent anticoagulant protein, annexin V, at the light and electron microscopic levels, using immunohistochemistry and an immunogold method. Annexin V was found to localize to the microvillar surface of the villous syncytiotrophoblasts. Isolated villous-derived trophoblasts were then utilized to evaluate the expression of annexin 1 protein mRNA in response to syncytialization in vitro, as well as to exposure to adenylate cyclase and protein kinase C agonists. Levels of immunoreactive annexin V released into the conditioned media and associated with cell protein were assessed by ELISA while levels of annexin V mRNA were evaluated by Northern analysis. No significant change in either media or cell-associated annexin V concentrations were detected over time in culture or in response to 1.5 mM 8-bromo-cyclic-adenosine-monophosphate (8-b-cAMP) or 0.15 nM phorbol ester myristic acid (PMA). These results indicate that annexin V is ideally positioned to inhibit intervillous thrombosis and maintain the fluidity of the intervillous circulation. Moreover, the absence of trophoblast annexin V regulation by intracellular second messenger regulators suggests that this crucial placental anticoagulant factor is constitutively produced.