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Cultured microvascular endothelial cells derived from the bovine corpus luteum possess NCAM-140
A Mayerhofer1, K Spanel-Borowski, S Watkins
1Abteilung Anatomie und Zellbiologie, Universität Ulm, Germany.
Abstract:
Previously, five phenotypically different, stable types of microvascular endothelial cells (MVE) were isolated from the bovine corpus and cultured successfully. We found that three out of these five types of MVE express the neural cell adhesion molecule (NCAM). As shown by immunocytochemistry, weak NCAM immunoreactivity occurred mainly in the perinuclear area of cell type 1. Monolayers of types 2 and 5 revealed heavy NCAM immunoreactivity, which was localized predominantly at the lateral cell surface outlining the contact zones of adjacent cells. In contrast, cell types 3 and 4 were not NCAM immunoreactive. Western blot analyses substantiated these results: While cell type 1 showed a weak immunoreactive band, cell types 2 and 5 displayed strong NCAM-immunoreactive bands of a molecular weight of approximately 140 kDa (NCAM-140), which was absent in cell types 3 and 4. These results reveal for the first time that NCAM can be expressed by cultured MVE and may serve in mediating endothelial cell contacts. Since luteal cells also express NCAM-140, this adhesion molecule could in addition be involved in the interactions of luteal cells with MVE.
Insights
Three bovine microvascular endothelial cell (MVE) types express neural cell adhesion molecule (NCAM). NCAM is localized at cell surfaces, potentially mediating endothelial cell contacts and interactions with luteal cells.
Area of Science:
- Cell Biology
- Endocrinology
- Immunology
Background:
- Five phenotypically distinct bovine microvascular endothelial cell (MVE) types were previously isolated and cultured.
- The expression of neural cell adhesion molecule (NCAM) in MVE was not well-characterized.
Purpose of the Study:
- To investigate the expression and localization of NCAM in cultured bovine MVE.
- To explore the potential role of NCAM in cell-cell interactions within the corpus luteum.
Main Methods:
- Isolation and culture of five stable MVE types from bovine corpus.
- Immunocytochemistry to detect NCAM expression and localization.
- Western blot analysis to confirm NCAM presence and molecular weight.
Main Results:
- Three of the five MVE types expressed NCAM.
- NCAM immunoreactivity varied: weak in cell type 1 (perinuclear), strong in types 2 and 5 (lateral cell surface).
- Western blots confirmed NCAM-140 expression in cell types 1, 2, and 5, with strongest bands in types 2 and 5.
Conclusions:
- Cultured MVE express NCAM, with distinct patterns depending on cell type.
- NCAM may play a role in mediating endothelial cell contacts.
- NCAM-140 expression in MVE suggests a potential role in interactions with luteal cells, which also express NCAM-140.