Apoptosis in human sperm: Its correlation with semen quality and the presence of leukocytes

G. Ricci, S. Perticarari, E. Fragonas, E. Giolo, S. Canova, C. Pozzobon, S. Guaschino, G. Presani

Research output: Contribution to journalArticle

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Abstract

Background: Apoptosis plays an important role in regulating spermatogenesis. However, the biological significance of apoptosis in ejaculated sperm is not yet clear. This study set out to investigate how apoptosis correlates with semen quality and the presence of seminal leukocytes. Methods: Fifty-seven semen samples from the male partners of infertile couples were classified as normal or abnormal according to World Health Organization guidelines. Flow cytometry was used to evaluate sperm populations and seminal leukocytes. Preliminary flow cytometry analysis using 6-carboxyfluoresceindiacetate (6-CFDA), which identifies live cells, and propidium iodide (PI), which stains only dead cells, was performed in order to pinpoint the sperm region accurately. Having thus gated the sperm population, bivariate Annexin V/PI analysis was then carried out in order to measure the percentage of apoptotic and necrotic sperm and the apoptotic index (the ratio between apoptotic:live sperm). Leukocytes were counted by the standard peroxidase test and by flow cytometry using monoclonal antibody (mAb) anti-CD45 or anti-CD53. Results: No significant differences in the apoptotic index and the percentage of live and apoptotic sperm were detected between the subjects with normal and abnormal semen. A significant inverse correlation between sperm concentration and the apoptotic index was observed only in the normal sperm group. There was no correlation between the concentration of leukocytes, detected either by peroxidase or by mAb anti-CD45 or anti-CD53, either with the percentage of apoptotic sperm or with the apoptotic index. In contrast, the ratio between CD45 positive leukocytes and sperm showed a significant correlation with the apoptotic index. A weaker correlation was found when leukocytes were counted by peroxidase, while no correlation was observed using mAb anti-CD53. Conclusions: Sperm apoptosis did not seem to be correlated with semen quality. In the absence of genito-urinary infection, one of the main functions of seminal leukocytes is probably to provide for the removal of apoptotic sperm.

Original languageEnglish
Pages (from-to)2665-2672
Number of pages8
JournalHuman Reproduction
Volume17
Issue number10
Publication statusPublished - Oct 1 2002

Fingerprint

Semen Analysis
Spermatozoa
Leukocytes
Apoptosis
Peroxidase
Flow Cytometry
Propidium
Monoclonal Antibodies
Semen
Annexin A5
Spermatogenesis
Population
Coloring Agents

Keywords

  • Apoptosis
  • Flow cytometry
  • Membrane phosphatidylserine
  • Seminal leukocytes
  • Sperm

ASJC Scopus subject areas

  • Physiology
  • Developmental Biology
  • Obstetrics and Gynaecology
  • Reproductive Medicine

Cite this

Ricci, G., Perticarari, S., Fragonas, E., Giolo, E., Canova, S., Pozzobon, C., ... Presani, G. (2002). Apoptosis in human sperm: Its correlation with semen quality and the presence of leukocytes. Human Reproduction, 17(10), 2665-2672.

Apoptosis in human sperm : Its correlation with semen quality and the presence of leukocytes. / Ricci, G.; Perticarari, S.; Fragonas, E.; Giolo, E.; Canova, S.; Pozzobon, C.; Guaschino, S.; Presani, G.

In: Human Reproduction, Vol. 17, No. 10, 01.10.2002, p. 2665-2672.

Research output: Contribution to journalArticle

Ricci, G, Perticarari, S, Fragonas, E, Giolo, E, Canova, S, Pozzobon, C, Guaschino, S & Presani, G 2002, 'Apoptosis in human sperm: Its correlation with semen quality and the presence of leukocytes', Human Reproduction, vol. 17, no. 10, pp. 2665-2672.
Ricci G, Perticarari S, Fragonas E, Giolo E, Canova S, Pozzobon C et al. Apoptosis in human sperm: Its correlation with semen quality and the presence of leukocytes. Human Reproduction. 2002 Oct 1;17(10):2665-2672.
Ricci, G. ; Perticarari, S. ; Fragonas, E. ; Giolo, E. ; Canova, S. ; Pozzobon, C. ; Guaschino, S. ; Presani, G. / Apoptosis in human sperm : Its correlation with semen quality and the presence of leukocytes. In: Human Reproduction. 2002 ; Vol. 17, No. 10. pp. 2665-2672.
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abstract = "Background: Apoptosis plays an important role in regulating spermatogenesis. However, the biological significance of apoptosis in ejaculated sperm is not yet clear. This study set out to investigate how apoptosis correlates with semen quality and the presence of seminal leukocytes. Methods: Fifty-seven semen samples from the male partners of infertile couples were classified as normal or abnormal according to World Health Organization guidelines. Flow cytometry was used to evaluate sperm populations and seminal leukocytes. Preliminary flow cytometry analysis using 6-carboxyfluoresceindiacetate (6-CFDA), which identifies live cells, and propidium iodide (PI), which stains only dead cells, was performed in order to pinpoint the sperm region accurately. Having thus gated the sperm population, bivariate Annexin V/PI analysis was then carried out in order to measure the percentage of apoptotic and necrotic sperm and the apoptotic index (the ratio between apoptotic:live sperm). Leukocytes were counted by the standard peroxidase test and by flow cytometry using monoclonal antibody (mAb) anti-CD45 or anti-CD53. Results: No significant differences in the apoptotic index and the percentage of live and apoptotic sperm were detected between the subjects with normal and abnormal semen. A significant inverse correlation between sperm concentration and the apoptotic index was observed only in the normal sperm group. There was no correlation between the concentration of leukocytes, detected either by peroxidase or by mAb anti-CD45 or anti-CD53, either with the percentage of apoptotic sperm or with the apoptotic index. In contrast, the ratio between CD45 positive leukocytes and sperm showed a significant correlation with the apoptotic index. A weaker correlation was found when leukocytes were counted by peroxidase, while no correlation was observed using mAb anti-CD53. Conclusions: Sperm apoptosis did not seem to be correlated with semen quality. In the absence of genito-urinary infection, one of the main functions of seminal leukocytes is probably to provide for the removal of apoptotic sperm.",
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