AQUAPORIN GENE EXPRESSION IN HUMAN FETAL MEMBRANE

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。羊水是正常妊娠的重要伴生物，是胎儿运动、生长发育所必需的。羊水过少，或房颤体积减少，发生在所有妊娠的8%到38%。房颤液体的膜内吸收途径(跨羊膜)最近被认为是房颤吸收的关键调节途径，对房颤容量的动态平衡有重要作用。然而，跨羊膜吸收水分的潜在分子和细胞机制仍不清楚。自从近十年前水通道蛋白被发现以来，水通道蛋白在水跨细胞膜运输中的作用越来越受到人们的重视。房颤的IM吸收(羊膜和绒毛膜上的水流向胎儿血管)可能是通过水通道调节的(S)。然而，在已鉴定的11种哺乳动物水通道蛋白中，很少有人在胎膜中进行检测。我们已经证明了AQP8在人和绵羊羊膜、绒毛和胎盘中的表达。这些结果表明，AQP8或其他水通道可能为液体跨胎膜运输提供了一条途径，有助于房颤量的稳态。本项目的目的是研究人胎膜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。将对正常足月妊娠的人胎盘和胎膜进行研究。剖宫产后，立即解剖羊膜、绒毛、胎盘和脐带，并将其冷冻在液氮中。组织将储存在零下70摄氏度，直到进一步分析。使用Trizol试剂(Life Technologies，Gaithersburg，MD)从这些组织中提取总RNA，用于RT-PCR。RT-PCR将用于检测水通道蛋白基因在这些组织中的表达。我们将在分娩时使用从受试者身上丢弃的人胎盘和胎膜。不会对受试者做任何事情。

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.