Toward an integrative and predictive sperm quality analysis in Bos taurus

Area of Science:

• Reproductive Biology
• Animal Science
• Biotechnology

Background:

• Current sperm quality analysis methods lack integration for simultaneous multi-parametric evaluation.
• There is a need for advanced techniques to assess sperm subpopulations cell by cell.

Purpose of the Study:

• To develop and validate a novel multi-parametric fluorescent test for comprehensive sperm analysis.
• To discriminate sperm subpopulations based on labeling patterns and motility characteristics.

Main Methods:

• Utilized cryopreserved Holstein bull semen samples.
• Performed computer-assisted sperm analysis (CASA-mot) for motility.
• Assessed membrane integrity using acridine orange-propidium iodide.
• Employed the ISAS®3Fun kit for multi-parametric analysis.

Main Results:

• The ISAS®3Fun kit successfully discriminated sperm subpopulations by membrane and acrosomal integrity, motility, and morphology.
• Identified live spermatozoa exhibiting capacitation signs like hyperactivated motility.
• Demonstrated correlations between intact plasma membrane/acrosome and higher motility.
• Observed static or weakly motile sperm with intact plasmalemma but damaged acrosomes.

Conclusions:

• The ISAS®3Fun kit is an integrated method advancing sperm quality analysis.
• This method shows potential for improving fertility predictions in bulls.
• Highlights the importance of assessing multiple sperm parameters concurrently.