Cytoskeletal filaments and associated proteins during restricted trophoblast invasion in bovine placentomes: light and transmission electron microscopy and RT-PCR.
Trophoblast cell migration is unusual in epitheliochorial placentae but occurs in placentomes of cows as "restricted" trophoblast invasion of binucleated trophoblast giant cells (TGC). Migration may be induced by integrin binding to the extracellular matrix initiating two pathways: (1) conformational changes of the actin cytoskeleton induced by an accumulation of its associated proteins and (2) integrin-dependent phosphorylation of various protein kinases. In cow placentomes, actin, its associated proteins (alpha-actinin, vinculin) and a key protein kinase of the signal transduction cascade (phosphorylated mitogen-activated protein kinase, pMAPK) were localized by immunogold-silver enhancement and immunoperoxidase staining at the light- and transmission electron-microscopical levels. Findings were confirmed by amplification of specific mRNA transcripts by reverse transcriptase/polymerase chain reaction. Actin and alpha-actinin were co-localized apically in mononuclear trophoblast cells, along the cytoplasmic membrane of TGC and apically in maternal crypt cells. The actin and alpha-actinin immunoreaction occurred as a band of electron-dense particles beneath the cytoplasmic membrane. Vinculin labelling was membrane-associated in TGC and in fetal and maternal endothelial cells. MAPK was observed as nuclear clusters in both kinds of trophoblast cells and was less dense in single uterine epithelial cells. Most MAPK immunoreactivity was detected in the nuclei of the trophoblast epithelium but was also sometimes membrane-associated in the cytoplasm. Thus, actin, alpha-actinin, MAPK and vinculin may be involved in the regulation of TGC migration. "Restricted" trophoblast invasion could serve as a model for invasive processes.
Lang
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CY
Department of Veterinary Anatomy, Histology and Embryology, Justus Liebig University, Frankfurter Strasse 98, 35392 Giessen, Germany. Carola.Y.Lang@vetmed.uni-giessen.de
Hallack
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S
Leiser
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R
Pfarrer
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D000199:Actins / Q000235:genetics / Q000378:metabolism; D000818:Animals; D002417:Cattle; D002465:Cell Movement / Q000502:physiology; D003599:Cytoskeleton / Q000378:metabolism* / Q000648:ultrastructure*; D005260:Female; D007124:Immunoenzyme Techniques; D008840:Microfilament Proteins / Q000378:metabolism* / Q000648:ultrastructure*; D008854:Microscopy, Electron; D020929:Mitogen-Activated Protein Kinase Kinases / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D011247:Pregnancy; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014327:Trophoblasts / Q000378:metabolism* / Q000648:ultrastructure*; D016596:Vinculin / Q000235:genetics / Q000378:metabolism
D000199:Actins; D008840:Microfilament Proteins; D012333:RNA, Messenger; D016596:Vinculin; D020929:Mitogen-Activated Protein Kinase Kinases