Identification of WNT/{beta}-CATENIN signaling pathway components in human cumulus cells |
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| Authors: | Wang Hong-Xing; Tekpetey Francis R; Kidder Gerald M |
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| Institution: | 1Department of Physiology and Pharmacology, The University of Western Ontario, London, ON, Canada N6A 5C1
2Department of Obstetrics and Gynaecology, The University of Western Ontario, London, ON, Canada N6A 5C1
3Reproductive Endocrinology and Infertility Program, London Health Sciences Centre, 339 Windermere Road, London, ON, Canada N6A 5A5
4Department of Paediatrics, The University of Western Ontario, London, ON, Canada N6A 5C1
5 Children’s Health Research Institute, 800 Commissioners Road East, London, ON, Canada N6C 2V5 |
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| Abstract: | Signaling via the conserved WNT/β-CATENIN pathway controlsdiverse developmental processes. To explore its potential rolein the ovary, we investigated the expression of WNTs, frizzled(FZD) receptors and other pathway components in human cumuluscells obtained from oocytes collected for in vitro fertilization.Proteins were detected in cultured cells using immunofluorescencemicroscopy. Protein–protein interactions were analyzedby means of immunoprecipitation. WNT2, FZD2, FZD3 and FZD9 wereidentified but WNT1, WNT4 and FZD4 were not detected. WNT2 isco-expressed with FZD2, FZD3 and FZD9. Co-immunoprecipitationusing WNT2 antibody demonstrated that WNT2 interacts with bothFZD3 and FZD9, but only FZD9 antibody precipitated WNT2. Wealso identified DVL (disheveled), AXIN, GSK-3β (glycogensynthase kinase-3β) and β-CATENIN. β-CATENINis concentrated in the plasma membranes. DVL co-localizes withFZD9 and AXIN in the membranes, but GSK-3β has little co-localizationwith AXIN and β-CATENIN. Interestingly, β-CATENINis highly co-localized with FZD9 and AXIN. CDH1 (E-cadherin)was also detected in the plasma membranes and cytoplasm, co-localizedwith β-CATENIN, and CDH1 antibody precipitated β-CATENIN.The results suggest that WNT2 could act through its receptorFZD9 to regulate the β-CATENIN pathway in cumulus cells,recruiting β-CATENIN into plasma membranes and promotingthe formation of adherens junctions involving CDH1. |
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| Keywords: | ovarian follicle/paracrine signaling/folliculogenesis/gene expression/signal transduction |
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