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Equine CRISP3 modulates interaction between spermatozoa and polymorphonuclear neutrophils.
1Department of Animal Sciences, College of Agriculture and Life Sciences, College of Veterinary Medicine, University of Florida, Gainesville, Florida, USA.
Biology of Reproduction
|March 11, 2011
Summary
Equine seminal plasma contains a protein, CRISP3, that suppresses the binding of sperm to neutrophils. This finding suggests CRISP3 regulates sperm removal from the mare's reproductive tract.
Area of Science:
- Reproductive immunology
- Protein biochemistry
Background:
- Equine spermatozoa trigger a uterine inflammatory response involving neutrophils.
- Seminal plasma proteins influence sperm-neutrophil interactions.
- The specific protein responsible for modulating this interaction remained unidentified.
Purpose of the Study:
- To isolate and identify a protein in equine seminal plasma that inhibits sperm-neutrophil binding.
- To elucidate the role of this protein in regulating the inflammatory response in the mare's reproductive tract.
Main Methods:
- Proteins from pooled stallion seminal plasma were precipitated and fractionated using ammonium sulfate precipitation and Sephacryl S200 chromatography.
- Active fractions were further purified by ion-exchange chromatography (DEAE Sephadex).
- Protein identification was achieved through SDS-PAGE, Western blotting, and sequence analysis; functional assays used flow cytometry to quantify PMN-sperm binding.
Main Results:
- A protein suppressing polymorphonuclear neutrophil (PMN)/sperm binding was isolated and identified as equine CRISP3.
- Equine seminal plasma and purified CRISP3 significantly reduced PMN/sperm binding compared to control media (63.09% ± 8.67% vs. 2.06% ± 2.02%, P < 0.0001).
- Flow cytometry confirmed the suppressive effect of CRISP3 on PMN-sperm interactions.
Conclusions:
- Equine seminal plasma contains CRISP3, a protein that effectively suppresses the binding of neutrophils to spermatozoa.
- CRISP3 likely plays a crucial role in regulating the clearance of spermatozoa from the female reproductive tract, modulating the post-mating inflammatory response.
