title: Expression and cellular localisation of chloride intracellular channel 3 in human placenta and fetal membranes.
abstract: Chloride channels regulate the movement of a major cellular anion and are involved in fundamental processes that are critical for cell viability. Regulation of intracellular chloride is achieved by multiple classes of channel proteins. One class of putative channels are the chloride intracellular channel (CLIC) family. Evidence suggests that several CLICs are expressed in human placenta, although their roles in this tissue are not certain. Northern blot analysis has shown that CLIC3 is highly expressed in placenta relative to other human tissues; however, its cellular distribution is not known. This study used microarray expression profiling to clarify which CLICs are expressed in human placenta and RT-PCR, Western blot and immunohistochemistry to determine the expression pattern of CLIC3 in human placenta and fetal membranes. Placentas and fetal membranes were obtained from term pregnancies after delivery and placental tissue was obtained from first trimester following either chorionic villous sampling or elective pregnancy termination. Trophoblast cells were isolated from first trimester and term placentas and placental endothelial cells were isolated from term placentas. Microarray expression profiling identified high expression of mRNA for CLICs 1, 3 and 4 in the isolated first trimester and term trophoblast cells. High mRNA expression in the isolated endothelial cells was also found for CLICs 1 and 4, but not CLIC3. Low expression was found for CLIC5 in all three types of isolated cells. RT-PCR confirmed that CLIC3 mRNA was expressed in trophoblast cells at both gestational ages, but was not present in endothelial cells. CLIC3 mRNA was also identified in whole placental extracts at both gestational ages and in term amnion and choriodecidua. Immunohistochemistry using a chicken anti-human CLIC3 antibody localised strong CLIC3-specific staining to the syncytiotrophoblast and villous cytotrophoblast cells in both first trimester and term placentas, and weaker staining in extravillous trophoblast cells in first trimester. In fetal membranes at term strong CLIC3-specific staining was localised to chorionic trophoblast cells, with weaker staining in amniotic epithelial and decidual cells. It was previously shown that chloride uptake was increased into cells that had been transfected with CLIC3. CLIC3 may facilitate chloride ion movement and the regulation of cellular processes associated with the movement of chloride in the placental and fetal membrane cells in which it is expressed.
authors.affiliation: Department of Pharmacology, Monash University, Clayton, Vic. 3800, Australia.
authors.forename: T T
authors.initials: TT
authors.lastname: Money
authors.forename: R G
authors.initials: RG
authors.lastname: King
authors.forename: M H
authors.initials: MH
authors.lastname: Wong
authors.forename: J L
authors.initials: JL
authors.lastname: Stevenson
authors.forename: B
authors.initials: B
authors.lastname: Kalionis
authors.forename: J J H M
authors.initials: JJ
authors.lastname: Erwich
authors.forename: M A
authors.initials: MA
authors.lastname: Huisman
authors.forename: A
authors.initials: A
authors.lastname: Timmer
authors.forename: U
authors.initials: U
authors.lastname: Hiden
authors.forename: G
authors.initials: G
authors.lastname: Desoye
authors.forename: N M
authors.initials: NM
authors.lastname: Gude
mesh_terms: D018118:Chloride Channels / Q000235:genetics*; D017931:DNA Primers; D005321:Extraembryonic Membranes / Q000166:cytology / Q000502:physiology*; D005260:Female; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D007150:Immunohistochemistry; D020411:Oligonucleotide Array Sequence Analysis; D010920:Placenta / Q000166:cytology / Q000502:physiology*; D011247:Pregnancy; D011261:Pregnancy Trimester, First; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: C117019:CLIC3 protein, human; D018118:Chloride Channels; D017931:DNA Primers; D012313:RNA