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An acute-phase protein as a regulator of sperm survival in the bovine oviduct: alpha 1-acid-glycoprotein impairs neutrophil phagocytosis of sperm in vitro.

Liu J, Marey MA, Kowsar R, Hambruch N, Shimizu T, Haneda S, Matsui M, Sasaki M, Hayakawa H, Pfarrer C, Miyamoto A - J. Reprod. Dev. (2014)

Bottom Line: AGP mRNA is expressed in extrahepatic organs, such as the lung, kidney, spleen, lymph node, uterus, and ovary.The AGP gene was expressed in cultured bovine oviduct epithelial cells (BOECs) and AGP protein was detected in oviduct fluid.The desialylation of AGP eliminated these suppressive effects of AGP on PMN.

View Article: PubMed Central - PubMed

Affiliation: Graduate School of Animal and Food Hygiene, Obihiro University of Agriculture and Veterinary Medicine, Hokkaido 080-8555, Japan.

ABSTRACT
We have previously shown that polymorphonuclear neutrophils (PMNs) are present in bovine oviduct fluid under physiological conditions, and that the oviduct provides a microenvironment that protects sperm from phagocytosis by PMNs. Alpha 1-acid glycoprotein (AGP) is a major acute-phase protein produced mainly in the liver that has immunomodulatory functions. AGP mRNA is expressed in extrahepatic organs, such as the lung, kidney, spleen, lymph node, uterus, and ovary. Therefore, in this study, we investigated, 1) the local production of AGP in the bovine oviduct, 2) the effect of AGP on the phagocytic activity of PMNs for sperm and superoxide production and 3) the impact of AGP desialylation on the PMN phagocytosis of sperm. The AGP gene was expressed in cultured bovine oviduct epithelial cells (BOECs) and AGP protein was detected in oviduct fluid. Preexposure of PMNs to AGP at physiological levels impaired PMN phagocytosis for sperm and superoxide generation. The desialylation of AGP eliminated these suppressive effects of AGP on PMN. Scanning electron microscopy revealed that AGP drastically reduced the formation of DNA-based neutrophil extracellular traps (NETs) for sperm entanglement. Additionally, AGP dose-dependently stimulated BOECs to produce prostaglandin E2 (PGE2) which has been shown to partially contribute to the regulation of sperm phagocytosis in the bovine oviduct. AGP and PGE2 at concentrations detected in the oviducts additively suppressed sperm phagocytosis by PMNs. These results provide evidence that locally produced AGP may be involved in protecting sperm from phagocytosis by PMNs in the bovine oviduct.

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(a) AGP gene expression in PMNs, BOECs, and the liver. (b) The AGP concentrations per oviduct flush during theestrous cycle. (preovulatory, n = 4, postovulatory, n = 5, and mid-luteal stage, n = 5) (c) Percentage of PMNphagocytosis for sperm treated to induce capacitation in vitro. PMNs were incubated for 4 h inculture medium supplemented with 0, 1, 10, or 100 ng/ml AGP and then subjected to a 1-h phagocytosis assay.Numerical values are presented as means ± SEM of three experiments. The different letters indicate significantdifferences between treatments at P < 0.05. (d) The effect of AGP on percentage of superoxide production by PMNsundergoing in vitro phagocytosis of sperm treated to induce capacitation. Numerical values arepresented as means ± SEM of four experiments. The different letters indicate significant differences between themarked treatments at P < 0.05.
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fig_001: (a) AGP gene expression in PMNs, BOECs, and the liver. (b) The AGP concentrations per oviduct flush during theestrous cycle. (preovulatory, n = 4, postovulatory, n = 5, and mid-luteal stage, n = 5) (c) Percentage of PMNphagocytosis for sperm treated to induce capacitation in vitro. PMNs were incubated for 4 h inculture medium supplemented with 0, 1, 10, or 100 ng/ml AGP and then subjected to a 1-h phagocytosis assay.Numerical values are presented as means ± SEM of three experiments. The different letters indicate significantdifferences between treatments at P < 0.05. (d) The effect of AGP on percentage of superoxide production by PMNsundergoing in vitro phagocytosis of sperm treated to induce capacitation. Numerical values arepresented as means ± SEM of four experiments. The different letters indicate significant differences between themarked treatments at P < 0.05.

Mentions: Our results demonstrate local gene expression of AGP by cultured BOECs in vitro that was well comparable tothat for the liver and PMNs (Fig. 1aFig. 1.


An acute-phase protein as a regulator of sperm survival in the bovine oviduct: alpha 1-acid-glycoprotein impairs neutrophil phagocytosis of sperm in vitro.

Liu J, Marey MA, Kowsar R, Hambruch N, Shimizu T, Haneda S, Matsui M, Sasaki M, Hayakawa H, Pfarrer C, Miyamoto A - J. Reprod. Dev. (2014)

(a) AGP gene expression in PMNs, BOECs, and the liver. (b) The AGP concentrations per oviduct flush during theestrous cycle. (preovulatory, n = 4, postovulatory, n = 5, and mid-luteal stage, n = 5) (c) Percentage of PMNphagocytosis for sperm treated to induce capacitation in vitro. PMNs were incubated for 4 h inculture medium supplemented with 0, 1, 10, or 100 ng/ml AGP and then subjected to a 1-h phagocytosis assay.Numerical values are presented as means ± SEM of three experiments. The different letters indicate significantdifferences between treatments at P < 0.05. (d) The effect of AGP on percentage of superoxide production by PMNsundergoing in vitro phagocytosis of sperm treated to induce capacitation. Numerical values arepresented as means ± SEM of four experiments. The different letters indicate significant differences between themarked treatments at P < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4219990&req=5

fig_001: (a) AGP gene expression in PMNs, BOECs, and the liver. (b) The AGP concentrations per oviduct flush during theestrous cycle. (preovulatory, n = 4, postovulatory, n = 5, and mid-luteal stage, n = 5) (c) Percentage of PMNphagocytosis for sperm treated to induce capacitation in vitro. PMNs were incubated for 4 h inculture medium supplemented with 0, 1, 10, or 100 ng/ml AGP and then subjected to a 1-h phagocytosis assay.Numerical values are presented as means ± SEM of three experiments. The different letters indicate significantdifferences between treatments at P < 0.05. (d) The effect of AGP on percentage of superoxide production by PMNsundergoing in vitro phagocytosis of sperm treated to induce capacitation. Numerical values arepresented as means ± SEM of four experiments. The different letters indicate significant differences between themarked treatments at P < 0.05.
Mentions: Our results demonstrate local gene expression of AGP by cultured BOECs in vitro that was well comparable tothat for the liver and PMNs (Fig. 1aFig. 1.

Bottom Line: AGP mRNA is expressed in extrahepatic organs, such as the lung, kidney, spleen, lymph node, uterus, and ovary.The AGP gene was expressed in cultured bovine oviduct epithelial cells (BOECs) and AGP protein was detected in oviduct fluid.The desialylation of AGP eliminated these suppressive effects of AGP on PMN.

View Article: PubMed Central - PubMed

Affiliation: Graduate School of Animal and Food Hygiene, Obihiro University of Agriculture and Veterinary Medicine, Hokkaido 080-8555, Japan.

ABSTRACT
We have previously shown that polymorphonuclear neutrophils (PMNs) are present in bovine oviduct fluid under physiological conditions, and that the oviduct provides a microenvironment that protects sperm from phagocytosis by PMNs. Alpha 1-acid glycoprotein (AGP) is a major acute-phase protein produced mainly in the liver that has immunomodulatory functions. AGP mRNA is expressed in extrahepatic organs, such as the lung, kidney, spleen, lymph node, uterus, and ovary. Therefore, in this study, we investigated, 1) the local production of AGP in the bovine oviduct, 2) the effect of AGP on the phagocytic activity of PMNs for sperm and superoxide production and 3) the impact of AGP desialylation on the PMN phagocytosis of sperm. The AGP gene was expressed in cultured bovine oviduct epithelial cells (BOECs) and AGP protein was detected in oviduct fluid. Preexposure of PMNs to AGP at physiological levels impaired PMN phagocytosis for sperm and superoxide generation. The desialylation of AGP eliminated these suppressive effects of AGP on PMN. Scanning electron microscopy revealed that AGP drastically reduced the formation of DNA-based neutrophil extracellular traps (NETs) for sperm entanglement. Additionally, AGP dose-dependently stimulated BOECs to produce prostaglandin E2 (PGE2) which has been shown to partially contribute to the regulation of sperm phagocytosis in the bovine oviduct. AGP and PGE2 at concentrations detected in the oviducts additively suppressed sperm phagocytosis by PMNs. These results provide evidence that locally produced AGP may be involved in protecting sperm from phagocytosis by PMNs in the bovine oviduct.

Show MeSH
Related in: MedlinePlus