| Autor: |
Brady, H.A., Blanchard, T.L., Evans, J.W., Varner, D.D., Bruemmer, J.E., Day, W., Schwab, C.A., Risek, B., Gilula, N.B., Burghardt, R.C. |
| Zdroj: |
Biology of Reproduction; January 1995, Vol. 52 Issue: 1 p507-514, 8p |
| Abstrakt: |
Gap junction expression in the endometrial epithelium of cycling and early pregnant mares was examined by electron microscopy, immunocytochemistry, and laser cytometry. Endometrial biopsies were procured per vaginum from mares during diestrus, estrus, and early pregnancy (Days 12,14, 15, 16, 18, and 23). One biopsy sample from each collection was immediately fixed for electron microscopy, and a second sample was divided for tissue culture and for immunocytochemistry. Ultrastructural analysis demonstrated the presence of gap junctions in the sub-apical membranes of epithelial cells from uterine glands and luminal epithelium. Small gap junctional contacts were also detected infrequently in the basolateral aspect of uterine gland epithelial cells. Screening of endometrial tissues with antibodies to connexin proteins α1(Cx43), β2(Cx32), and β2(Cx26) by indirect immunofluorescence revealed that α1is a predominant connexin in equine endometrial glandular and luminal epithelium that is concentrated along the subapical membrane and corresponds to the prominent gap junctions seen at the ultrastructural level. Both ultrastructure and immunocytochemistry indicated that gap junctions between epithelial cells were present at all stages examined. Laser cytometric analysis of intercellular communication in isolated uterine glandular and luminal epithelial cells by a fluorescence recovery following photobleaching (FRAP) assay verified the communication competence of these cells regardless of day of cycle or stage of early pregnancy sampled. Immunocytochemical analysis of primary cultures confirmed extensive α1staining along appositional contact between cells. The presence of α1in equine endometrial epithelial cells is in contrast to observations in the rat, where the β2connexin is present. Extensive metabolic coupling may potentially play a role in both embryo recognition and nutritional support required for the survival of the embryo in the mare. |
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