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Involvement of cortactin and phosphotyrosine proteins in cell–cell contact formation in cultured bovine corneal endothelial cells

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Abstract

Phosphotyrosine proteins involvement, particularly cortactin, was studied in cell–cell contacts of cultured bovine corneal endothelial (BCE) cells. These proteins, including α-catenin, vinculin and cortactin, are localized at cell–cell contacts separate from the cortical actin ring. Approximately 50% of cortactin isoforms p80 and p85 were associated with the Triton-insoluble fraction while phosphotyrosine proteins were in the soluble fraction. Disruption of cell–cell contacts by EDTA treatment was associated with a decrease in cortactin isoforms p80 (26%) and p85 (57%). Cortactin isoform p85 was phosphorylated at Y466, expressed in reattaching cells and associated with the Triton-soluble fraction, whereas cortactin isoform p80 was phosphorylated at Y421 and associated with the Triton-insoluble fraction. In sub-confluent cultures, pY421-cortactin was localized at the leading edge and pY466-cortactin at a perinuclear area. In confluent cultures both pY466- and pY421-cortactin isoforms were localized at the cell–cell contacts. In conclusion, in BCE cells, the most prominent appearance of cortactin was at the cell–cell contacts separate from the cortical actin ring. Isoform p80 was phosphorylated at Y421 and associated with the Triton-insoluble fraction and isoform p85 was phosphorylated at Y466 and associated with the Triton-insoluble fraction.

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Acknowledgments

This study was supported by a grant from the Claire and Amedee Marateir Institute for the Study of Blindness and Visual Disorders. The authors thank Yair Andegeko, Tel-Aviv University, Tel-Aviv, for his contribution to the immunofluorescence studies and to Mark Tarshis, Ph.D. The Hebrew University of Jerusalem, Jerusalem, for his devoted technical support to the confocal microscopy studies. This work was conducted in partial fulfillment of the requirements for Ph.D. degree of Lily Kredy-Farhan, Sackler faculty of medicine, Tel Aviv University, Tel Aviv, Israel.

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Authors and Affiliations

  1. Goldschleger Eye Research Institute and the Department of Human Molecular Genetics and Biochemistry, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel

    Lily Kredy-Farhan, Shlomo Kotev-Emeth & Naphtali Savion

  2. Goldschleger Eye Research Institute, Sheba Medical Center, 52621, Tel Hashomer, Israel

    Naphtali Savion

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  1. Lily Kredy-Farhan
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  2. Shlomo Kotev-Emeth
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  3. Naphtali Savion
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Correspondence to Naphtali Savion.

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Kredy-Farhan, L., Kotev-Emeth, S. & Savion, N. Involvement of cortactin and phosphotyrosine proteins in cell–cell contact formation in cultured bovine corneal endothelial cells. Histochem Cell Biol 129, 193–202 (2008). https://doi.org/10.1007/s00418-007-0357-8

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