Hum. Reprod. Advance Access originally published online on January 29, 2004
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Human Reproduction, Vol. 19, No. 3, 607-610,
March 2004
© 2004 European Society of Human Reproduction and Embryology
Apoptosis and necrosis in human ejaculated spermatozoa
1 Department of Biochemistry and Molecular Biology, University of Granada, Campus Universitario ‘Fuentenueva’, 18701 Granada, 2 MAR&Gen, Molecular Assisted Reproduction and Genetics, Gracia 36, 18002 Granada, Spain and 3 Laboratoire d’Eylau, 55 rue Saint Didier, 75116 Paris, France
4 To whom correspondence should be addressed at: MAR&Gen, Gracia 36, 18002 Granada, Spain. e-mail: cmendoza{at}ugr.es
BACKGROUND: Features of both apoptosis and necrosis have been reported in ejaculated human spermatozoa. This study examines the relative contribution of these two modes of cell death to the demise of these terminally differentiated cells. METHODS: Sperm fractions were prepared from aliquots of semen samples from young normozoospermic donors by simple washing from seminal plasma, by discontinuous density gradient centrifugation or by swim-up technique. They were subsequently incubated in vitro at 37°C for 24 h. Sperm motility, viability, and the presence of two apoptotic markers, phosphatidylserine externalization (annexin-V binding) and DNA fragmentation (TUNEL), were examined before incubation and again after 4 and 24 h of incubation. RESULTS: The swim-up technique was the most efficient in terms of the recovery of viable, motile and non-apoptotic spermatozoa, followed by density gradient centrifugation and finally simple washing. No changes in the parameters tested were observed after 4 h of incubation, but a significant decrease in sperm motility and viability was detected after 24 h irrespective of the sperm preparation technique employed. However, these changes were not accompanied by any increase in the incidence of spermatozoa showing markers of an active apoptotic process. CONCLUSIONS: Healthy human ejaculated spermatozoa appear incapable of initiating apoptosis, at least under in vitro conditions.
Key words: apoptosis/cell death/DNA fragmentation/phosphatidylserine externalization/spermatozoa
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