Shengbiao Wang1, Jiexiong Chen, Katherine T Au, Michael G Ross. 1. Department of Obstetrics and Gynecology, Harbor-UCLA Research and Education Institute, School of Medicine, University of California, Los Angeles 90502, USA. sheng@gcrc.rei.edu
Abstract
OBJECTIVE: Water absorption across the fetal chorioamniotic membranes is a critical regulatory pathway for amniotic fluid volume homeostasis. Aquaporins are cell membrane proteins that significantly enhance membrane permeability to water by acting as water channels. We recently demonstrated that aquaporin 8 is expressed in human amnion, chorion, and placenta. Thus, aquaporin 8 expression represents a molecular mechanism of amniotic water absorption through intramembranous pathways. The current study sought to determine whether aquaporin 8 is expressed in human amnion-derived cell culture and to explore its regulation by second messenger cyclic adenosine monophosphate. STUDY DESIGN: Human amnion-derived WISH cells were cultured. Total RNA was isolated and reverse transcriptase-polymerase chain reaction was used to determine aquaporin 8 gene expression. To determine the effect of cyclic adenosine monophosphate on aquaporin 8 expression, WISH cells were cultured in the presence of either monobutyryl cyclic adenosine monophosphate or the cyclic adenosine monophosphate-elevating agent forskolin. Multiplex semiquantitative reverse transcriptase-polymerase chain reaction was carried out to quantify aquaporin 8 messenger RNA levels. RESULTS: Reverse transcriptase-polymerase chain reaction detected aquaporin 8 expression in WISH cells. After forskolin treatment for 2 hours, aquaporin 8 messenger RNA expression in WISH cells increased 4-fold (P <.001). Stimulation of aquaporin 8 gene expression by colforsin was observed throughout the study period of 20 hours. Incubation of WISH cells with monobutyryl cyclic adenosine monophosphate resulted in a 2-fold increase in aquaporin 8 messenger RNA level (P <.001). However, stimulation of aquaporin 8 gene expression by monobutyryl cyclic adenosine monophosphate attenuated to baseline level after 20 hours of monobutyryl cyclic adenosine monophosphate treatment. CONCLUSION: The current study demonstrates the expression of aquaporin 8 water channel in human amnion-derived WISH cells and aquaporin 8 expression up-regulation by second messenger cyclic adenosine monophosphate. Aquaporin 8 messenger RNA demonstrates a relatively short biologic half-life in vitro, which renders its rapid responsiveness to regulation
OBJECTIVE:Water absorption across the fetal chorioamniotic membranes is a critical regulatory pathway for amniotic fluid volume homeostasis. Aquaporins are cell membrane proteins that significantly enhance membrane permeability to water by acting as water channels. We recently demonstrated that aquaporin 8 is expressed in human amnion, chorion, and placenta. Thus, aquaporin 8 expression represents a molecular mechanism of amniotic water absorption through intramembranous pathways. The current study sought to determine whether aquaporin 8 is expressed in human amnion-derived cell culture and to explore its regulation by second messenger cyclic adenosine monophosphate. STUDY DESIGN:Human amnion-derived WISH cells were cultured. Total RNA was isolated and reverse transcriptase-polymerase chain reaction was used to determine aquaporin 8 gene expression. To determine the effect of cyclic adenosine monophosphate on aquaporin 8 expression, WISH cells were cultured in the presence of either monobutyryl cyclic adenosine monophosphate or the cyclic adenosine monophosphate-elevating agent forskolin. Multiplex semiquantitative reverse transcriptase-polymerase chain reaction was carried out to quantify aquaporin 8 messenger RNA levels. RESULTS: Reverse transcriptase-polymerase chain reaction detected aquaporin 8 expression in WISH cells. After forskolin treatment for 2 hours, aquaporin 8 messenger RNA expression in WISH cells increased 4-fold (P <.001). Stimulation of aquaporin 8 gene expression by colforsin was observed throughout the study period of 20 hours. Incubation of WISH cells with monobutyryl cyclic adenosine monophosphate resulted in a 2-fold increase in aquaporin 8 messenger RNA level (P <.001). However, stimulation of aquaporin 8 gene expression by monobutyryl cyclic adenosine monophosphate attenuated to baseline level after 20 hours of monobutyryl cyclic adenosine monophosphate treatment. CONCLUSION: The current study demonstrates the expression of aquaporin 8water channel in human amnion-derived WISH cells and aquaporin 8 expression up-regulation by second messenger cyclic adenosine monophosphate. Aquaporin 8 messenger RNA demonstrates a relatively short biologic half-life in vitro, which renders its rapid responsiveness to regulation
Authors: Elizabeth M Jablonski; M Adrian Mattocks; Eugene Sokolov; Leonidas G Koniaris; Francis M Hughes; Nelson Fausto; Robert H Pierce; Iain H McKillop Journal: Cancer Lett Date: 2006-11-03 Impact factor: 8.679
Authors: Dawoon Jung; J Denry Sato; Joseph R Shaw; Bruce A Stanton Journal: Comp Biochem Physiol A Mol Integr Physiol Date: 2011-12-13 Impact factor: 2.320