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Journal of Assisted Reproduction and Genetics
Published in: Journal of Assisted Reproduction and Genetics 5/2015

01-05-2015 | Gamete Biology

Can apoptosis and necrosis coexist in ejaculated human spermatozoa during in vitro semen bacterial infection?

Authors: Monika Fraczek, Magdalena Hryhorowicz, Dariusz Gaczarzewicz, Anna Szumala-Kakol, Tomasz J. Kolanowski, Lothar Beutin, Maciej Kurpisz

Published in: Journal of Assisted Reproduction and Genetics | Issue 5/2015

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Abstract

Purpose

To evaluate whether ejaculated human spermatozoa undergo complete apoptosis or necrosis during experimental semen bacterial infection in vitro.

Methods

Apoptotic markers, including mitochondrial transmembrane potential (ΔΨm), phosphatidylserine (PS) externalization, and DNA fragmentation, have been detected simultaneously in ejaculated human sperm after their incubation with a known pathogenic (Escherichia coli), as well as with conditionally pathogenic bacterial strains (Staphylococcus haemolyticus, Bacteroides ureolyticus) and/or leukocytes. The ΔΨm and translocation of PS was evaluated using the JC-1 and Annexin V binding tests, respectively. A modified TUNEL assay with additional staining for sperm viability was used to detect the DNA fragmentation level.

Results

The exposure of ejaculated spermatozoa to bacterial strains was associated with a simultaneous decrease in the percentage of sperm with normal ΔΨm and an increase in the proportion of Annexin V-positive sperm. Additionally, in the presence of S. haemolyticus, B. ureolyticus and/or leukocytes, a significant increase in the percentage of live TUNEL-positive (apoptotic) as well as dead TUNEL-positive (necrotic) sperm cells was also observed.

Conclusions

The cellular death observed in spermatozoa in the presence of inflammatory mediators may be due to both apoptosis and necrosis. Here, we demonstrate for the first time that direct contact of conditionally pathogenic bacteria with ejaculated human sperm may play an even greater role in the promotion of apoptosis than in case of some pathogenic bacterial strains. These findings suggest that significant bacteriospermia and leukocytospermia may be direct causes of subfertility or additional negative factors worsening the prognosis of fertility in natural and assisted procreation.
Appendix
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Metadata
Title
Can apoptosis and necrosis coexist in ejaculated human spermatozoa during in vitro semen bacterial infection?
Authors
Monika Fraczek
Magdalena Hryhorowicz
Dariusz Gaczarzewicz
Anna Szumala-Kakol
Tomasz J. Kolanowski
Lothar Beutin
Maciej Kurpisz
Publication date
01-05-2015
Publisher
Springer US
Published in
Journal of Assisted Reproduction and Genetics / Issue 5/2015
Print ISSN: 1058-0468
Electronic ISSN: 1573-7330
DOI
https://doi.org/10.1007/s10815-015-0462-x

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