Focal Adhesion Kinase Is a Key Mediator of Human Trophoblast Development
Trophoblast differentiation during the first trimester of pregnancy involves cell proliferation and invasion and extracellular matrix (ECM) remodeling. Reports have indicated that, in a variety of cell types, processes such as proliferation, invasion, and ECM remodeling require the turnover of focal...
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| Veröffentlicht in: | Laboratory investigation 2001-11, Vol.81 (11), p.1469-1483 |
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| creator | MacPhee, Daniel James Mostachfi, Homa Han, Robin Lye, Stephen James Post, Martin Caniggia, Isabella |
| description | Trophoblast differentiation during the first trimester of pregnancy involves cell proliferation and invasion and extracellular matrix (ECM) remodeling. Reports have indicated that, in a variety of cell types, processes such as proliferation, invasion, and ECM remodeling require the turnover of focal adhesions mediated by a cytoplasmic tyrosine kinase named focal adhesion kinase (FAK). Therefore, in the present study we examined the expression and spatial localization of FAK during early human placental development. Immunocytochemical and immunoblot analysis showed that FAK and a focal adhesion-associated protein named paxillin were highly expressed between the 5th and 8th weeks of gestation, specifically in villous cytotrophoblast and extravillous trophoblast (EVT) cells. Activated FAK, phosphorylated on Tyr-397, colocalized with α5 integrin and matrix metalloproteinase-2 (MMP2) expression in EVT cells within a previously characterized intermediate, invasive-restrained region. FAK and paxillin expression dramatically decreased after 10 to 12 weeks of gestation coincident with increasing pO2 levels. Exposure of human villous explants of 5 to 8 weeks to a 3% O2 environment resulted in increased trophoblast outgrowth, cell proliferation, and detection of α5 integrin and MMP2, as well as increased activation of FAK in EVT cells compared with explants grown in a 20% O2 environment. To determine whether FAK was a key requisite for trophoblast differentiation, villous explants of 5 weeks gestation were grown in Matrigel in a 3% O2 environment and incubated with 20-mer antisense FAK oligonucleotides. A dramatic reduction of trophoblast outgrowth was observed in antisense-treated explants compared with missense and control cultures, and, in addition, cell proliferation and MMP2 activity in antisense-treated explants were dramatically reduced. These data suggest that FAK is a key kinase involved in early trophoblast cell differentiation and plays a role in regulating cell proliferation and motility during early placental development. |
| doi_str_mv | 10.1038/labinvest.3780362 |
| format | Article |
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Reports have indicated that, in a variety of cell types, processes such as proliferation, invasion, and ECM remodeling require the turnover of focal adhesions mediated by a cytoplasmic tyrosine kinase named focal adhesion kinase (FAK). Therefore, in the present study we examined the expression and spatial localization of FAK during early human placental development. Immunocytochemical and immunoblot analysis showed that FAK and a focal adhesion-associated protein named paxillin were highly expressed between the 5th and 8th weeks of gestation, specifically in villous cytotrophoblast and extravillous trophoblast (EVT) cells. Activated FAK, phosphorylated on Tyr-397, colocalized with α5 integrin and matrix metalloproteinase-2 (MMP2) expression in EVT cells within a previously characterized intermediate, invasive-restrained region. FAK and paxillin expression dramatically decreased after 10 to 12 weeks of gestation coincident with increasing pO2 levels. Exposure of human villous explants of 5 to 8 weeks to a 3% O2 environment resulted in increased trophoblast outgrowth, cell proliferation, and detection of α5 integrin and MMP2, as well as increased activation of FAK in EVT cells compared with explants grown in a 20% O2 environment. To determine whether FAK was a key requisite for trophoblast differentiation, villous explants of 5 weeks gestation were grown in Matrigel in a 3% O2 environment and incubated with 20-mer antisense FAK oligonucleotides. A dramatic reduction of trophoblast outgrowth was observed in antisense-treated explants compared with missense and control cultures, and, in addition, cell proliferation and MMP2 activity in antisense-treated explants were dramatically reduced. These data suggest that FAK is a key kinase involved in early trophoblast cell differentiation and plays a role in regulating cell proliferation and motility during early placental development.</description><identifier>ISSN: 0023-6837</identifier><identifier>EISSN: 1530-0307</identifier><identifier>DOI: 10.1038/labinvest.3780362</identifier><identifier>PMID: 11706056</identifier><identifier>CODEN: LAINAW</identifier><language>eng</language><publisher>New York: Elsevier Inc</publisher><subject>Antisense Elements (Genetics) ; Biological and medical sciences ; Cell Differentiation - physiology ; Cell Division - physiology ; Cell Movement - physiology ; Cells, Cultured ; Cytoskeletal Proteins - analysis ; Cytoskeletal Proteins - genetics ; Embryology: invertebrates and vertebrates. Teratology ; Fetal membranes ; Fetus - cytology ; Focal Adhesion Kinase 1 ; Focal Adhesion Protein-Tyrosine Kinases ; Fundamental and applied biological sciences. Psychology ; Gene Expression Regulation, Developmental - drug effects ; Gene Expression Regulation, Developmental - physiology ; Gene Expression Regulation, Enzymologic - drug effects ; Gene Expression Regulation, Enzymologic - physiology ; General aspects. Development. Fetal membranes ; Humans ; Laboratory Medicine ; Medicine ; Medicine & Public Health ; Oxygen - pharmacology ; Pathology ; Paxillin ; Phosphoproteins - analysis ; Phosphoproteins - genetics ; Protein-Tyrosine Kinases - analysis ; Protein-Tyrosine Kinases - genetics ; Trophoblasts - enzymology</subject><ispartof>Laboratory investigation, 2001-11, Vol.81 (11), p.1469-1483</ispartof><rights>2001 United States & Canadian Academy of Pathology</rights><rights>The United States and Canadian Academy of Pathology, Inc. 2001</rights><rights>2002 INIST-CNRS</rights><rights>Copyright Nature Publishing Group Nov 2001</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c580t-6f28bf4039c4954d548087953ef375694af7eff994e69c18211480f486e5d5993</citedby><cites>FETCH-LOGICAL-c580t-6f28bf4039c4954d548087953ef375694af7eff994e69c18211480f486e5d5993</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=14152850$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/11706056$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>MacPhee, Daniel James</creatorcontrib><creatorcontrib>Mostachfi, Homa</creatorcontrib><creatorcontrib>Han, Robin</creatorcontrib><creatorcontrib>Lye, Stephen James</creatorcontrib><creatorcontrib>Post, Martin</creatorcontrib><creatorcontrib>Caniggia, Isabella</creatorcontrib><title>Focal Adhesion Kinase Is a Key Mediator of Human Trophoblast Development</title><title>Laboratory investigation</title><addtitle>Lab Invest</addtitle><addtitle>Lab Invest</addtitle><description>Trophoblast differentiation during the first trimester of pregnancy involves cell proliferation and invasion and extracellular matrix (ECM) remodeling. Reports have indicated that, in a variety of cell types, processes such as proliferation, invasion, and ECM remodeling require the turnover of focal adhesions mediated by a cytoplasmic tyrosine kinase named focal adhesion kinase (FAK). Therefore, in the present study we examined the expression and spatial localization of FAK during early human placental development. Immunocytochemical and immunoblot analysis showed that FAK and a focal adhesion-associated protein named paxillin were highly expressed between the 5th and 8th weeks of gestation, specifically in villous cytotrophoblast and extravillous trophoblast (EVT) cells. Activated FAK, phosphorylated on Tyr-397, colocalized with α5 integrin and matrix metalloproteinase-2 (MMP2) expression in EVT cells within a previously characterized intermediate, invasive-restrained region. FAK and paxillin expression dramatically decreased after 10 to 12 weeks of gestation coincident with increasing pO2 levels. Exposure of human villous explants of 5 to 8 weeks to a 3% O2 environment resulted in increased trophoblast outgrowth, cell proliferation, and detection of α5 integrin and MMP2, as well as increased activation of FAK in EVT cells compared with explants grown in a 20% O2 environment. To determine whether FAK was a key requisite for trophoblast differentiation, villous explants of 5 weeks gestation were grown in Matrigel in a 3% O2 environment and incubated with 20-mer antisense FAK oligonucleotides. A dramatic reduction of trophoblast outgrowth was observed in antisense-treated explants compared with missense and control cultures, and, in addition, cell proliferation and MMP2 activity in antisense-treated explants were dramatically reduced. 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Reports have indicated that, in a variety of cell types, processes such as proliferation, invasion, and ECM remodeling require the turnover of focal adhesions mediated by a cytoplasmic tyrosine kinase named focal adhesion kinase (FAK). Therefore, in the present study we examined the expression and spatial localization of FAK during early human placental development. Immunocytochemical and immunoblot analysis showed that FAK and a focal adhesion-associated protein named paxillin were highly expressed between the 5th and 8th weeks of gestation, specifically in villous cytotrophoblast and extravillous trophoblast (EVT) cells. Activated FAK, phosphorylated on Tyr-397, colocalized with α5 integrin and matrix metalloproteinase-2 (MMP2) expression in EVT cells within a previously characterized intermediate, invasive-restrained region. FAK and paxillin expression dramatically decreased after 10 to 12 weeks of gestation coincident with increasing pO2 levels. Exposure of human villous explants of 5 to 8 weeks to a 3% O2 environment resulted in increased trophoblast outgrowth, cell proliferation, and detection of α5 integrin and MMP2, as well as increased activation of FAK in EVT cells compared with explants grown in a 20% O2 environment. To determine whether FAK was a key requisite for trophoblast differentiation, villous explants of 5 weeks gestation were grown in Matrigel in a 3% O2 environment and incubated with 20-mer antisense FAK oligonucleotides. A dramatic reduction of trophoblast outgrowth was observed in antisense-treated explants compared with missense and control cultures, and, in addition, cell proliferation and MMP2 activity in antisense-treated explants were dramatically reduced. These data suggest that FAK is a key kinase involved in early trophoblast cell differentiation and plays a role in regulating cell proliferation and motility during early placental development.</abstract><cop>New York</cop><pub>Elsevier Inc</pub><pmid>11706056</pmid><doi>10.1038/labinvest.3780362</doi><tpages>15</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Antisense Elements (Genetics) Biological and medical sciences Cell Differentiation - physiology Cell Division - physiology Cell Movement - physiology Cells, Cultured Cytoskeletal Proteins - analysis Cytoskeletal Proteins - genetics Embryology: invertebrates and vertebrates. Teratology Fetal membranes Fetus - cytology Focal Adhesion Kinase 1 Focal Adhesion Protein-Tyrosine Kinases Fundamental and applied biological sciences. Psychology Gene Expression Regulation, Developmental - drug effects Gene Expression Regulation, Developmental - physiology Gene Expression Regulation, Enzymologic - drug effects Gene Expression Regulation, Enzymologic - physiology General aspects. Development. Fetal membranes Humans Laboratory Medicine Medicine Medicine & Public Health Oxygen - pharmacology Pathology Paxillin Phosphoproteins - analysis Phosphoproteins - genetics Protein-Tyrosine Kinases - analysis Protein-Tyrosine Kinases - genetics Trophoblasts - enzymology |
| title | Focal Adhesion Kinase Is a Key Mediator of Human Trophoblast Development |
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