This subproject is one of many research subprojects utilizing theresources provided by a Center grant funded by NIH/NCRR. The subproject andinvestigator (PI) may have received primary funding from another NIH source,and thus could be represented in other CRISP entries. The institution listed isfor the Center, which is not necessarily the institution for the investigator.Amniotic fluid (AF) is an essential accompaniment of normal pregnancy, necessary for fetal movement, growth and development. Oligohydramnios (OH), or reduced AF volume, occurs in 8 to 38% of all pregnancies. The intramembranous pathway of AF fluid absorption (across the amniotic membrane) has recently been recognized as a critical regulatory path for AF resorption, contributing importantly to AF volume homeostasis. Yet the underlying molecular and cellular mechanisms for water absorption across the amniotic membranes remain unknown. Since the discovery of AQP water channels about a decade ago, AQPs are increasingly recognized to play an important role in water transport across cell membranes. IM resorption of AF (water flow across the amnion and perhaps chorion, to the fetal vasculature) is potentially mediated via AQP water channel(s). However, of the eleven identified mammalian AQPs, few have been examined in fetal membranes. We have demonstrated AQP8 expression in both human and ovine amnion, chorion and placenta. These results suggest that AQP8, or other water channels may provide a pathway for fluid transport across fetal membranes, contributing to AF volume homeostasis. The purpose of the project is to investigate the aquaporins gene expression in human fetal membrane to further determine which water channels contribute to the regulation of amniotic fluid resorption. Human placenta and fetal membranes from normal term pregnancy will be studied. Immediately upon cesarean section delivery, amnion, chorion, placenta, and umbilical cord will be dissected and frozen in liquid nitrogen. Tissues will be stored at -70C until further analysis. Total RNA will be isolated from these tissues using Trizol reagent (Life Technologies, Gaithersburg, MD) for RT-PCR. RT-PCR will be used to detect the aquaporins gene expression in these tissues. We will use the discarded human placenta and fetal membranes from the subject upon delivery. Nothing will be done to the subjects.
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