Apoptosis in spermatozoa of infertile men, clinical correlations

The methods for evaluation of male infertility include not
only routine investigations, standardized by the WHO, but also
complementary techniques, developed over the last years, in
order to improve the predictive value of seminal analysis for
natural conception and assisted reproduction. With reference
to these new methods, studies suggest that sperm with certain
levels of DNA fragmentation serve as a strong predictor of
reduced male fertility. We studied subjects who underwent
seminal fluid evaluation, because of an infertility condition, at
the Department of Biomedical Sciences of the University of
Sassari.The samples collected by masturbation were evaluated
according to the World Health Organisation (1999).The samples
was washed twice in PBS and cytocentrifuged for 10 min
at 1800 rpm on polylysine-coated slides that were fixed in
methanol at room temperature. The apoptosis was evaluated
using the TUNEL (In Situ Cell Death Detection Kit,
Fluorescein, Roche, Cat.No. 1 684 795). At fluorescent miscroscopy are counted at least 300 cells. Quantitative evaluation
of apoptosis by the TUNEL method confirmed that apoptosis
did not seem to be correlated with sperm concentration or morphology; however, we found a higher apoptotic rate in semen
from patients affected by andrologic diseases, such as varicocele,
than from those with alteration of semen characteristics.
Apoptosis analysis might be used in infertile patients in order
to understand the etiology of unexplained infertility and to
improve therapeutic effectiveness.