Hypoxia inhibits differentiation of lineage-specific Rcho-1 trophoblast giant cells.
Defects in placental development lead to pregnancies at risk for miscarriage and intrauterine growth retardation and are associated with preeclampsia, a leading cause of maternal death and premature birth. In preeclampsia, impaired placental formation has been associated with alterations in a specific trophoblast lineage, the invasive trophoblast cells. In this study, an RT-PCR Trophoblast Gene Expression Profile previously developed by our laboratory was utilized to examine the lineage-specific gene expression of the rat Rcho-1 trophoblast cell line. Our results demonstrated that Rcho-1 cells represent an isolated, trophoblast population committed to the giant cell lineage. RT-PCR analysis revealed that undifferentiated Rcho-1 cells expressed trophoblast stem cell marker, Id2, and trophoblast giant cell markers. On differentiation, Rcho-1 cells downregulated Id2 and upregulated Csh1, a marker of the trophoblast giant cell lineage. Neither undifferentiated nor differentiated Rcho-1 cells expressed spongiotrophoblast marker Tpbpa or labyrinthine markers Esx1 and Tec. Differentiating Rcho-1 cells in hypoxia did not alter the expression of lineage-specific markers; however, hypoxia did inhibit the downregulation of the trophoblast stem cell marker Id2. Differentiation in hypoxia also blocked the induction of CSH1 protein. In addition, hypoxia inhibited stress fiber formation and abolished the induction of palladin, a protein associated with stress fiber formation and focal adhesions. Thus, Rcho-1 cells can be maintained as a proliferative, lineage-specific cell line that is committed to the trophoblast giant cell lineage on differentiation in both normoxic and hypoxic conditions; however, hypoxia does inhibit aspects of trophoblast giant cell differentiation at the molecular, morphological, and functional levels.
Department of Neuroscience, Cell Biology, and Physiology, Wright State University Boonshoft School of Medicine, Dayton, Ohio 45435, USA.
Amy D
AD
Gultice
Kaisa L
KL
Selesniemi
Thomas L
TL
Brown
D000818:Animals; D002454:Cell Differentiation / Q000235:genetics / Q000502:physiology*; D015687:Cell Hypoxia / Q000502:physiology*; D002460:Cell Line; D019070:Cell Lineage / Q000235:genetics / Q000502:physiology; D049109:Cell Proliferation; D003598:Cytoskeletal Proteins / Q000032:analysis / Q000235:genetics / Q000502:physiology; D015536:Down-Regulation; D005260:Female; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental / Q000235:genetics / Q000502:physiology; D051797:Inhibitor of Differentiation Protein 2 / Q000032:analysis / Q000235:genetics / Q000502:physiology; D010750:Phosphoproteins / Q000032:analysis / Q000235:genetics / Q000502:physiology; D010920:Placenta / Q000473:pathology; D010928:Placental Lactogen / Q000032:analysis / Q000235:genetics / Q000502:physiology; D010929:Placentation; D011257:Pregnancy Proteins / Q000032:analysis / Q000235:genetics / Q000502:physiology; D011505:Protein-Tyrosine Kinases / Q000032:analysis / Q000235:genetics / Q000502:physiology; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014157:Transcription Factors / Q000032:analysis / Q000235:genetics / Q000502:physiology; D014327:Trophoblasts / Q000737:chemistry / Q000166:cytology* / Q000502:physiology*; D015854:Up-Regulation
D003598:Cytoskeletal Proteins; C497451:Id2 protein, rat; D051797:Inhibitor of Differentiation Protein 2; C485874:LOC103693936; D010750:Phosphoproteins; D011257:Pregnancy Proteins; C411574:Tpbpa protein, rat; D014157:Transcription Factors; C085699:placental lactogen I, rat; D010928:Placental Lactogen; D011505:Protein-Tyrosine Kinases; C513330:Tec protein, rat