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Purification of a factor from human peritoneal fluid that is able to immobilize spermatozoa
G A Soldati1, A Piffaretti-Yanez, G Medici
1Andrology Laboratory, La Carità Hospital, Locarno, Switzerland.
Abstract:
Human peritoneal fluid has been claimed to influence sperm motility. This report gives evidence for the presence in midcycle peritoneal fluid of a protein-bound, lipidic (hydrophobic) component able to immobilize spermatozoa as a function of time. This component was extracted from molecular weight-sieving and ion-exchange/high pressure liquid chromatography (HPLC)-purified peritoneal fluid fractions by either chloroform/methanol or charcoal treatments; resuspension of the chloroform/methanol extract with BWW-buffer and subsequent testing on spermatozoa resulted in sperm immobilization. Sequential or step-down chromatographic procedures (molecular weight-sieving-->cation-exchange-->anion-exchange HPLC separations of native peritoneal fluid) and extensive dialysis against double distilled water allowed the purification of the sperm immobilizing factor, as evidenced by the shorter incubation times necessary for sperm immobilization. Furthermore, the active fraction was found to immobilize spermatozoa without affecting its viability. Separation of the chloroform/methanol extracted immobilizing fraction on thin layer chromatography under conditions for phospholipid detection allowed the identification of a characteristic band which, after re-extraction, was found to be the sperm immobilizing substance. This factor does not contain choline, ethanolamine or serine. These results suggest that some lipidic peritoneal fluid components may influence sperm motility.
Insights
Midcycle peritoneal fluid contains a lipidic component that immobilizes sperm over time. This hydrophobic factor affects sperm motility without impacting viability, suggesting a role for peritoneal fluid lipids in reproductive processes.
Area of Science:
- Reproductive Biology
- Biochemistry
- Sperm Physiology
Background:
- Human peritoneal fluid's influence on sperm motility is debated.
- Identifying factors within peritoneal fluid that affect sperm function is crucial for understanding reproductive health.
Purpose of the Study:
- To investigate the presence and nature of a sperm immobilizing factor in human midcycle peritoneal fluid.
- To characterize the biochemical properties of this sperm immobilizing component.
Main Methods:
- Extraction of peritoneal fluid components using chloroform/methanol and charcoal treatments.
- Purification using molecular weight-sieving, ion-exchange, and high-pressure liquid chromatography (HPLC).
- Sperm immobilization assays and thin-layer chromatography for component identification.
Main Results:
- A protein-bound, lipidic (hydrophobic) component was identified in midcycle peritoneal fluid that immobilizes spermatozoa.
- The active factor was purified through sequential chromatography and dialysis.
- The immobilizing substance affects sperm motility without compromising sperm viability and is identified as a phospholipid lacking choline, ethanolamine, or serine.
Conclusions:
- Lipidic components in human peritoneal fluid can influence sperm motility.
- A specific hydrophobic factor in peritoneal fluid acts as a sperm immobilizing agent.
- This finding contributes to understanding the biochemical interactions affecting sperm function in the female reproductive tract.