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Identification and Characterization of Protein Glycosylation using Specific Endo- and Exoglycosidases
Published on: December 26, 2011
Identification and characterization of glycosylation-dependent cell adhesion molecule 1-like protein expression in
T E Spencer1, F F Bartol, F W Bazer
1Center for Animal Biotechnology, Albert B. Alkek Institute of Biosciences and Technology, Texas A&M University, College Station, Texas 77843-2471, USA. tspencer@ansc.tamu.edu
Insights
Glycosylation-dependent cell adhesion molecule 1 (GlyCAM-1) is found in the sheep uterus during pregnancy. This protein may play a role in interactions between the conceptus and mother during early pregnancy.
Area of Science:
- Reproductive Biology
- Immunology
- Glycobiology
Background:
- Glycosylation-dependent cell adhesion molecule 1 (GlyCAM-1) is an endothelial glycoprotein crucial for lymphocyte extravasation in lymph nodes.
- Its function in the uterus, particularly during pregnancy, is not well understood.
Purpose of the Study:
- To investigate the presence and distribution of GlyCAM-1-like protein in the ovine uterus during the estrous cycle and pregnancy.
- To explore the potential role of GlyCAM-1 in conceptus-maternal interactions during peri-implantation.
Main Methods:
- Immunohistochemical analysis using rabbit anti-rat GlyCAM-1 IgG on ovine endometrial and conceptus tissues.
- Western blot analysis of endometrial extracts and uterine flushings to identify immunoreactive GlyCAM-1 protein.
Main Results:
- GlyCAM-1 expression in the endometrial luminal epithelium and glandular epithelium varied with the estrous cycle and pregnancy status.
- Immunoreactive GlyCAM-1 was detected in the conceptus trophectoderm during mid-to-late gestation.
- A 45 kDa GlyCAM-1-like protein was identified in endometrial extracts and uterine flushings, with levels increasing during early pregnancy.
Conclusions:
- A GlyCAM-1-like protein is expressed in the ovine uterus and may be secreted by the endometrial epithelium and/or conceptus trophectoderm.
- The distribution patterns suggest GlyCAM-1-like protein involvement in conceptus-maternal interactions during peri-implantation in sheep.
Abstract:
Glycosylation-dependent cell adhesion molecule 1 (GlyCAM-1) is an endothelial glycoprotein secreted in lymph nodes that serves as a ligand for leukocyte cell surface selectin and mediates lymphocyte extravasation. In the present studies, rabbit anti-rat GlyCAM-1 IgG was used in immunochemical analyses of GlyCAM-1-like protein in the ovine uterus. In cyclic ewes, GlyCAM-1 expression increased in the endometrial luminal epithelium (LE) and shallow glandular epithelium (cGE) between Days 1 and 5 and then decreased between Days 11 and 15. In pregnant ewes, GlyCAM-1 in the LE and cGE was low on Days 11 and 13, increased on Day 15, and was abundant on Days 17 and 19. Immunoreactive GlyCAM-1 was also detected in the conceptus trophectoderm on Days 13-19. Staining for GlyCAM-1 in the smooth muscle of the vasculature and myometrium was constitutive, and no staining was detected in the stroma. An immunoreactive protein of approximately 45 kDa was identified in endometrial extracts and uterine flushings from cyclic and pregnant ewes. In pregnant ewes, the relative amount of immunoreactive GlyCAM-1 in uterine flushings was low on Days 11 and 13 but high on Days 15 and 17. Results suggest that a GlyCAM-1-like protein may be a secretory product of the endometrial epithelium and/or conceptus trophectoderm. Patterns of distribution observed for immunoreactive GlyCAM-1-like protein in the endometrial epithelium, combined with proposed functions for lymphoid GlyCAM-1, suggest that this mucin glycoprotein may be involved in conceptus-maternal interactions during the periimplantation period of pregnancy in sheep.
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