title: Characterization of morphological and cytoskeletal changes in trophoblast cells induced by insulin-like growth factor-I.
abstract: IGF-I and IGF-II were appeared to play major roles in the adhesive and migratory events that are considered to be crucial in the implantation process. The purpose of this study was to determine the effects of IGF-I on trophoblast adhesion to extracellular matrix. Trophoblast cells obtained from early gestation at artificial abortion were incubated with the indicated doses of IGF-I at the indicated times. Trophoblast cells were treated with IGF-I in the presence or absence of RGD peptide and an antibody against alpha-subunit of IGF-I receptor (alphaIR3). Morphometric and morphological changes were studied using light and electron microscopy. Furthermore, vinculin, actin stress fibers, phosphorylated focal adhesion kinase (FAK), phosphotyrosine, and paxillin were immunolocalized in trophoblast cells after IGF-I treatment in the presence or absence of alphaIR3. Immunoprecipitation and anti-phosphotyrosine immunoblotting were carried out to detect the phosphorylated FAK and phosphorylated paxillin contents of the IGF-I-treated and untreated trophoblast cells. The results showed that IGF-I promoted trophoblast adhesion to fibronectin substrate in a time- and dose-dependent manner, and addition of RGD peptide and alphaIR3 monoclonal antibody abolished the effects of IGF-I in these cells. Morphological studies exhibited an increase in the lamellipodia formation upon IGF-I treatment, and confocal images of immunofluorescent staining revealed localization of phosphorylated FAK, paxillin, and vinculin at focal adhesions as well as redistribution of actin microfilaments and formation of actin stress fibers inside the cell. Western blotting, using antiphosphotyrosine demonstrated proteins with molecular masses of 125 kDa (FAK) and 68 kDa (paxillin) present in the IGF-I-treated cells, which were lacking in the control groups. In conclusion, these findings suggest that IGF-I can stimulate lamellipodia formation and promote adhesion of trophoblast cells to extracellular matrix by activating their adhesion molecules that must be activated within the implantation window.
authors.affiliation: Department of Obstetrics and Gynecology, Kyorin University School of Medicine, Mitaka, Tokyo 181-8611, Japan.
authors.forename: Maryam
authors.initials: M
authors.lastname: Kabir-Salmani
authors.forename: Shigetatsu
authors.initials: S
authors.lastname: Shiokawa
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Akimoto
authors.forename: Habib
authors.initials: H
authors.lastname: Hasan-Nejad
authors.forename: Keiji
authors.initials: K
authors.lastname: Sakai
authors.forename: Shinya
authors.initials: S
authors.lastname: Nagamatsu
authors.forename: Ken
authors.initials: K
authors.lastname: Sakai
authors.forename: Yukio
authors.initials: Y
authors.lastname: Nakamura
authors.forename: Ahmad
authors.initials: A
authors.lastname: Hosseini
authors.forename: Mitsutoshi
authors.initials: M
authors.lastname: Iwashita
mesh_terms: D002448:Cell Adhesion / Q000187:drug effects; D002478:Cells, Cultured; D003598:Cytoskeletal Proteins / Q000378:metabolism; D003599:Cytoskeleton / Q000502:physiology*; D004305:Dose-Response Relationship, Drug; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D007334:Insulin-Like Growth Factor I / Q000008:administration & dosage / Q000494:pharmacology*; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D051419:Paxillin; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D014327:Trophoblasts / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure*
chemical_list: D003598:Cytoskeletal Proteins; C496061:PXN protein, human; D051419:Paxillin; D010750:Phosphoproteins; D007334:Insulin-Like Growth Factor I; D011505:Protein-Tyrosine Kinases; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; C496050:PTK2 protein, human