Expression of aquaporin 8 and its up-regulation by cyclic adenosine monophosphate in human WISH cells.

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

journal_name

Am J Obstet Gynecol

authors

Wang S,Chen J,Au KT,Ross MG

doi

10.1067/mob.2003.214

subject

Has Abstract

pub_date

2003-04-01 00:00:00

pages

997-1001

issue

4

eissn

0002-9378

issn

1097-6868

pii

S0002937802715342

journal_volume

188

pub_type

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