AQUAPORIN GENE EXPRESSION IN HUMAN FETAL MEMBRANE

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。羊水（AF）是正常妊娠的基本伴随物，是胎儿运动、生长和发育所必需的。羊水过少（OH）或AF体积减少发生在所有妊娠的8%至38%中。AF液体吸收的膜内途径（穿过羊膜）最近被认为是AF再吸收的关键调节途径，对AF体积稳态有重要贡献。然而，水通过羊膜吸收的潜在分子和细胞机制仍然未知。自10多年前发现AQP水通道以来，人们越来越认识到AQP在水跨膜转运中的重要作用。AF的IM再吸收（水流通过羊膜和可能的绒毛膜到达胎儿血管系统）可能通过AQP水通道介导。然而，11个确定的哺乳动物水通道蛋白，很少有被检查在胎膜。我们已经证明了AQP 8在人和羊的羊膜，绒毛膜和胎盘中的表达。这些结果表明，AQP 8或其他水通道可能提供了一个途径，液体运输通过胎膜，有助于AF体积稳态。本研究的目的是研究人胎儿膜中水通道蛋白基因的表达，以进一步确定哪些水通道参与了羊水吸收的调节。将研究正常足月妊娠的人胎盘和胎膜。剖宫产分娩后，立即解剖羊膜、绒毛膜、胎盘和脐带，并在液氮中冷冻。将组织储存在-70 ℃下，直至进一步分析。使用Trizol试剂（Life Technologies，盖瑟斯堡，MD）从这些组织中分离总RNA进行RT-PCR。采用RT-PCR方法检测水通道蛋白基因在这些组织中的表达。 我们将使用受试者分娩时丢弃的人胎盘和胎膜。不会对受试者做任何事。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for 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.