Sperm Apoptosis in Fresh and Cryopreserved Bull Semen Detected by Flow Cytometry and Its Relationship with Fertility

The present study was conducted to detect sperm apoptosis in fresh and frozen semen and to determine its relationship with bull fertility. Three ejaculates were collected from five breeding bulls with different fertility levels and were cryopreserved using standard methods. Two flow cytometric metho...

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Bibliographic Details
Published in:Biology of reproduction 2002-02, Vol.66 (2), p.354-360
Main Authors: ANZAR, Muhammad, LIWEI HE, BUHR, Mary M, KROETSCH, Thomas G, PAULS, Karl P
Format: Article
Language:English
Subjects:
Animals
Annexin A5 - physiology
Apoptosis - drug effects
Apoptosis - physiology
Biological and medical sciences
Cattle
Cryopreservation
Fertility - drug effects
Fertility - physiology
Flow Cytometry
In Situ Nick-End Labeling
In Vitro Techniques
Male
Propidium
Semen - cytology
Semen - drug effects
Semen - physiology
Semen Preservation
Spermatozoa - drug effects
Spermatozoa - physiology
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Summary:The present study was conducted to detect sperm apoptosis in fresh and frozen semen and to determine its relationship with bull fertility. Three ejaculates were collected from five breeding bulls with different fertility levels and were cryopreserved using standard methods. Two flow cytometric methods were employed to measure apoptosis: an assay for phosphatidylserine (PS) translocation across the plasma membranes using fluorescein-labeled Annexin V and propidium iodide (PI), and an assay for nicked DNA using bromodeoxyuridine (BrdU), terminal deoxynucleotidyl transferase, and fluorescein-labeled anti-BrdU monoclonal antibody. Both assays showed that fresh sperm contained 10%–20% apoptotic sperm. Significant differences in the percentage of apoptotic sperm were observed among the bulls. Cryopreservation induced translocation of PS to the outer leaflet of the plasma membrane and caused most of the necrotic cells in fresh sperm to disintegrate. Bull fertility was significantly related to the percentage of necrotic or viable sperm in fresh semen as detected by the Annexin V/PI assay, to the number of apoptotic sperm in fresh semen as detected by the TUNEL assay, and to the level of chromatin or DNA condensation as detected by PI staining. The present study suggests that the presence of apoptotic spermatozoa in fresh semen could be one of the reasons for poor fertility in breeding bulls.
ISSN:0006-3363
1529-7268
DOI:10.1095/biolreprod66.2.354