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Expression of a P-selectin ligand in zona pellucida of porcine oocytes and P-selectin on acrosomal membrane of porcine sperm cells. Potential implications for their involvement in sperm-egg interactions.

Geng JG, Raub TJ, Baker CA, Sawada GA, Ma L, Elhammer AP - J. Cell Biol. (1997)

Bottom Line: In addition, a search for a specific receptor for this ligand leads to the identification of P-selectin on the acrosomal membrane of porcine sperm cells.Moreover, porcine sperm cells were found to be capable of binding to human promyeloid cell line HL-60.Taken together, our findings implicate a potential role for the oocyte P-selectin ligand and the sperm P-selectin in porcine sperm-egg interactions.

View Article: PubMed Central - PubMed

Affiliation: Cell Biology and Inflammation Research, Pharmacia and Upjohn, Inc., Kalamazoo, Michigan 49001, USA. jian-guo.geng@am.pnu.com

ABSTRACT
The selectin family of cell adhesion molecules mediates initial leukocyte adhesion to vascular endothelial cells at sites of inflammation. O-glycan structural similarities between oligosaccharides from human leukocyte P-selectin glycoprotein ligand-1 (PSGL-1) and from zona pellucida glycoproteins of porcine oocytes indicate the possible existence of a P-selectin ligand in the zona pellucida. Here, using biochemical as well as morphological approaches, we demonstrate that a P-selectin ligand is expressed in the porcine zona pellucida. In addition, a search for a specific receptor for this ligand leads to the identification of P-selectin on the acrosomal membrane of porcine sperm cells. In vitro binding of porcine acrosome-reacted sperm cells to oocytes was found to be Ca2+ dependent and inhibitable with either P-selectin, P-selectin receptor-globulin, or leukocyte adhesion blocking antibodies against P-selectin and PSGL-1. Moreover, porcine sperm cells were found to be capable of binding to human promyeloid cell line HL-60. Taken together, our findings implicate a potential role for the oocyte P-selectin ligand and the sperm P-selectin in porcine sperm-egg interactions.

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Staining of porcine  zona pellucida with PSGL-1  peptide antibody. Zona pellucida of porcine oocytes  were stained with either (A)  PSGL-1 peptide antibody or  (B) preimmune IgG followed  by indirect immunoperoxidase EM as described above.  The outer surfaces of the  zona are shown. Bar, 2 μm.
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Figure 4: Staining of porcine zona pellucida with PSGL-1 peptide antibody. Zona pellucida of porcine oocytes were stained with either (A) PSGL-1 peptide antibody or (B) preimmune IgG followed by indirect immunoperoxidase EM as described above. The outer surfaces of the zona are shown. Bar, 2 μm.

Mentions: Using EM, the distribution of the P-selectin ligand in the zona pellucida of porcine oocytes was examined after labeling either with P-selectin followed by P-selectin antibody or with PSGL-1 peptide antibody. The experiments demonstrated that the P-selectin ligand was associated with membrane fragments and vesicles embedded throughout the matrix of the zona pellucida (Figs. 3 and 4). The specificities of these approaches were confirmed by the absence, or marked reduction, of peroxidase reaction product in control oocytes, where either P-selectin (Fig. 3 C) or P-selectin antibody (data not shown) was omitted, where P-selectin was incubated in the presence of 2 mM EDTA (Fig. 3 D), or where preimmune IgG was used (Fig. 4 B).


Expression of a P-selectin ligand in zona pellucida of porcine oocytes and P-selectin on acrosomal membrane of porcine sperm cells. Potential implications for their involvement in sperm-egg interactions.

Geng JG, Raub TJ, Baker CA, Sawada GA, Ma L, Elhammer AP - J. Cell Biol. (1997)

Staining of porcine  zona pellucida with PSGL-1  peptide antibody. Zona pellucida of porcine oocytes  were stained with either (A)  PSGL-1 peptide antibody or  (B) preimmune IgG followed  by indirect immunoperoxidase EM as described above.  The outer surfaces of the  zona are shown. Bar, 2 μm.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2139885&req=5

Figure 4: Staining of porcine zona pellucida with PSGL-1 peptide antibody. Zona pellucida of porcine oocytes were stained with either (A) PSGL-1 peptide antibody or (B) preimmune IgG followed by indirect immunoperoxidase EM as described above. The outer surfaces of the zona are shown. Bar, 2 μm.
Mentions: Using EM, the distribution of the P-selectin ligand in the zona pellucida of porcine oocytes was examined after labeling either with P-selectin followed by P-selectin antibody or with PSGL-1 peptide antibody. The experiments demonstrated that the P-selectin ligand was associated with membrane fragments and vesicles embedded throughout the matrix of the zona pellucida (Figs. 3 and 4). The specificities of these approaches were confirmed by the absence, or marked reduction, of peroxidase reaction product in control oocytes, where either P-selectin (Fig. 3 C) or P-selectin antibody (data not shown) was omitted, where P-selectin was incubated in the presence of 2 mM EDTA (Fig. 3 D), or where preimmune IgG was used (Fig. 4 B).

Bottom Line: In addition, a search for a specific receptor for this ligand leads to the identification of P-selectin on the acrosomal membrane of porcine sperm cells.Moreover, porcine sperm cells were found to be capable of binding to human promyeloid cell line HL-60.Taken together, our findings implicate a potential role for the oocyte P-selectin ligand and the sperm P-selectin in porcine sperm-egg interactions.

View Article: PubMed Central - PubMed

Affiliation: Cell Biology and Inflammation Research, Pharmacia and Upjohn, Inc., Kalamazoo, Michigan 49001, USA. jian-guo.geng@am.pnu.com

ABSTRACT
The selectin family of cell adhesion molecules mediates initial leukocyte adhesion to vascular endothelial cells at sites of inflammation. O-glycan structural similarities between oligosaccharides from human leukocyte P-selectin glycoprotein ligand-1 (PSGL-1) and from zona pellucida glycoproteins of porcine oocytes indicate the possible existence of a P-selectin ligand in the zona pellucida. Here, using biochemical as well as morphological approaches, we demonstrate that a P-selectin ligand is expressed in the porcine zona pellucida. In addition, a search for a specific receptor for this ligand leads to the identification of P-selectin on the acrosomal membrane of porcine sperm cells. In vitro binding of porcine acrosome-reacted sperm cells to oocytes was found to be Ca2+ dependent and inhibitable with either P-selectin, P-selectin receptor-globulin, or leukocyte adhesion blocking antibodies against P-selectin and PSGL-1. Moreover, porcine sperm cells were found to be capable of binding to human promyeloid cell line HL-60. Taken together, our findings implicate a potential role for the oocyte P-selectin ligand and the sperm P-selectin in porcine sperm-egg interactions.

Show MeSH
Related in: MedlinePlus