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La vitalité des spermatozoïdes

The viability of spermatozoa

Résumé

Malgré l’existence de nombreux moyens d’étude de la vitalité des spermatozoïdes, la plupart restent dans le domaine de la recherche biomédicale. Ce sont surtout les tests classiques, tels que le test à l’éosinenigrosine et le test de gonflement flagellaire en milieu hypo-osmolaire (HOST) qui permettent d’apprécier le pourcentage de vitalité des spermatozoïdes en spermiologie courante. Un résultat anormalement bas définit la nécrozoospermie, dont l’interprétation est souvent difficile, mais permet d’orienter le bilan masculin vers la recherche de certains contextes étiologiques, notamment celui des infections génitales chroniques.

L’étude de la vitalité des spermatozoïdes constitue également l’élément-clé dans le choix des spermatozoïdes susceptibles d’être utilisés en vue d’une micro-injection intracytoplasmique. Dans cette indication, seuls les tests en milieu hypo-osmolaire sont applicables. Une analyse individuelle de chaque spermatozoïde avec un temps de séjour minimal dans la solution hypo-osmolaire semble actuellement préférable.

L’exploration de la vitalité des spermatozoïdes concerne désormais également l’apoptose. Cette forme de mort cellulaire peut être mise en évidence dans les spermatozoïdes par différents moyens techniques et pose le problème de la sélection de spermatozoïdes utilisables en assistance médicale à la procréation selon des critères d’intégrité non seulement membranaire, mais aussi génomique.

Abstract

Most of the numerous techniques used to assess sperm viability only have research applications, while only two classical tests, i.e. eosin-Y and hypo-osmotic swelling test (HOST), are currently used in routine sperm analysis to determine the percentage of viable sperm.

A viability rate below 50% of living sperm defines necrozoospermia, a condition whose clinical significance is fairly difficult to assess as the mechanisms of sperm cell death are still poorly understood. However, even when a precise cause for necrozoospermia cannot be identified, abnormal viability requires further andrological investigations with particular emphasis on clinical and laboratory signs of chronic infection of the male reproductive tract.

Intracytoplasmic sperm injection (ICSI) can yield very good pregnancy rates, even in couples with the most severe forms of male infertility. However, when no motile sperm are available after sperm preparation, the outcome of ICSI is seriously impaired, probably because of a high risk of injecting dead sperm.

In these patients, sperm viability could therefore be assessed by the hypo-osmotic swelling test in order to select only viable sperm for ICSI. However, the long incubation time of sperm in the hypo-osmotic solution, as recommended in the classical HOST procedure, has been shown to be detrimental to the spermatozoa. A single sperm test able to assess the viability of each individual spermatozoon within microdroplets covered by mineral oil therefore seems to be preferable. This selection procedure is less suitable in the case of immotile frozen-thawed sperm, as viability does not appear to be reliably predicted by HOST in cryopreserved sperm.

Examination of sperm viability now also evaluates programmed cell death or apoptosis, as apoptotic alterations can be detected in spermatozoa by several techniques. The percentage of apoptotic sperm is correlated with deficient sperm parameters and poor outcome of assisted reproductive techniques. More effective selection procedures are therefore needed in order to identify spermatozoa not only with intact membranes but also with an intact genome to be used for ICSI.

References

  1. 1.

    AHMADI A., NG S.C.: The single sperm curling test, a modified hypo-osmotic swelling test, as a potential technique for the selection of viable sperm for intracytoplasmic sperm injection. Fertil. Steril., 1997, 68: 346–350.

    PubMed  Article  CAS  Google Scholar 

  2. 2.

    AITKEN R.J., GORDON E., HARKISS D.et al.: Relative impact of oxidative stress on the functional competence and genomic integrity of human spermatozoa. Biol. Reprod., 1998, 59: 1037–1046.

    PubMed  Article  CAS  Google Scholar 

  3. 3.

    BACHTELL N.E., CONAGHAN J., TUREK P.J.: The relative viability of human spermatozoa from the vas deferens, epididymis and testis before and after cryopreservation. Hum. Reprod., 1999, 14: 3048–3051.

    PubMed  Article  CAS  Google Scholar 

  4. 4.

    BARROSO G., MORSHEDI M., OEHNINGER S.: Analysis of DNA fragmentation, plasma membrane translocation of phosphatidylserine and oxidative stress in human spermatozoa. Hum. Reprod., 2000, 15: 1338–1344.

    PubMed  Article  CAS  Google Scholar 

  5. 5.

    BLOM E.: A one-minute live-dead sperm stain by means of eosin-nigrosin. Fertil. Steril., 1950, 1: 176.

    Google Scholar 

  6. 6.

    CASPER R.F., MERIANO J.S., JARVI K.A., COWAN L., LUCATO M.L.: The hypo-osmotic swelling test for selection of viable sperm for intracytoplasmic sperm injection in men with complete asthenozoospermia. Fertil. Steril., 1996, 65: 972–976.

    PubMed  CAS  Google Scholar 

  7. 7.

    CHAN P.J., TREDWAY D.R., PANG S.C., CORSELLI J., SU B.C.: Assessment of sperm for cryopreservation using the hyposmotic viability test. Fertil. Steril., 1992, 58: 841–844.

    PubMed  CAS  Google Scholar 

  8. 8.

    DONNELLY E.T., O’CONNELL M., McCLURE N., LEWIS S.E.M.: Differences in nuclear DNA fragmentation and mitochondrial integrity of semen and prepared human spermatozoa. Hum. Reprod., 2000, 15: 1552–1561.

    PubMed  Article  CAS  Google Scholar 

  9. 9.

    ESTEVES S.C., SHARMA R.K., THOMAS A.J., AGARWAL A.: Suitability of the hypo-osmotic swelling test for assessing the viability of cryopreserved sperm. Fertil. Steril., 1996, 66: 798–804.

    PubMed  CAS  Google Scholar 

  10. 10.

    GANDINI L., LOMBARDO F., PAOLI D.et al.: Study of apoptotic DNA fragmentation in human spermatozoa. Hum. Reprod., 2000, 15: 830–839.

    PubMed  Article  CAS  Google Scholar 

  11. 11.

    GARNER D.A., PINKEL D., JOHNSONL A., PACE M.M.: Assessment of spermatozoal function using dual fluorecscent staining and flow cytometric analyses. Biol. Reprod., 1986, 34: 127–138.

    PubMed  Article  CAS  Google Scholar 

  12. 12.

    GARNER D.L., JOHNSON L.A.: Viability assessment of mammalian sperm using SYBR-14 and propidium iodide. Biol. Reprod., 1995, 53: 276–284.

    PubMed  Article  CAS  Google Scholar 

  13. 13.

    HOSSAIN A.M., RIZK B., BARIK S., HUFF C., THORNEYCROFT I.H.: Time course of hypo-osmotic swellings of human spermatozoa: evidence of ordered transition between swelling subtypes. Hum. Reprod., 1998, 13: 1578–1583.

    PubMed  Article  CAS  Google Scholar 

  14. 14.

    JEYENDRAN R.S., VAN DER VEN H.H., PEREZPELAES B.G., CRABO B.G., ZANEVELD L.J.: Development of an assay to assess the functional integrity of the human sperm membrane and its relationship to other semen characteristics. J. Reprod. Fertil., 1984, 70: 219–228.

    PubMed  CAS  Article  Google Scholar 

  15. 15.

    LIN M.H., MORSHEDI M., SRISOMBUT C., NASSAR A., OENINGER S.: Plasma membrane integrity of cryopreserved human sperm: an investigation of the results of the hypoosmotic swelling test, the water test, and eosin-Y staining. Fertil. Steril., 1998, 70: 1148–1155.

    PubMed  Article  CAS  Google Scholar 

  16. 16.

    LIU J., NAGY JORIS H.et al.: Analysis of 76 total fertilization-failure cycles out of 2732 intracytoplasmic sperm injection cycles. Hum. Reprod., 1995, 10: 2630–2636.

    PubMed  CAS  Google Scholar 

  17. 17.

    LIU J., TSAI Y.L., KATZ E., COMPTON G., GARCIA J.E., BARAMKI T.A.: High fertilization rate obtained after intracytoplasmic sperm injection with 100% nonmotile spermatozoa selected by using a simple modified hypo-osmotic swelling test. Fertil. Steril., 1997, 68: 373–375.

    PubMed  Article  CAS  Google Scholar 

  18. 18.

    LOMEO A.M., GIAMBERSIO A.M.: Water-test: a simple method to assess sperm membrane integrity. Int. J. Androl., 1991, 14: 278–282.

    PubMed  Article  CAS  Google Scholar 

  19. 19.

    LOPES S., SUN J.G., JURISICOVA A., MERIANO J., CASPER R.F.: Sperm deoxyribonucleic acid fragmentation is increased in poor-quality semen samples and correlates with failed fertilization in intracytoplasmic sperm injection. Fertil. Steril., 1998, 69: 528–532.

    PubMed  Article  CAS  Google Scholar 

  20. 20.

    McLAUGHLIN E.A., FORD W.C.L., HULL M.G.R.: Effects of cryopreservation on the human sperm acrosome and its response to A23187. J. Reprod. Fertil., 1993, 99: 71–76.

    PubMed  CAS  Google Scholar 

  21. 21.

    NAGY Z., LIU J., JORIS H.et al.: The result of intracytoplasmic sperm injection is not related to any of the three basic sperm parameters. Hum. Reprod., 1995, 10: 1123–1129.

    PubMed  CAS  Google Scholar 

  22. 22.

    NDUWAYO L., BARTHELEMY C., LANSAC J., THARANNE M.J., LECOMTE P.: Conduite à tenir devant une nécrozoospermie. Contracept. Fertil. Sex., 1995, 23: 682–685.

    PubMed  CAS  Google Scholar 

  23. 23.

    OOSTERHUIS G.J., MULDER A.B., KALSBEEK-BATENBURG E., LAMBALK C.B., SCHOEMAKER J., VERMES I.: Measuring apoptosis in human spermatozoa: a biological assay for semen quality? Fertil. Steril., 2000, 74: 245–250.

    PubMed  Article  CAS  Google Scholar 

  24. 24.

    SAKKAS D., MARIETHOS K., St JOHN J.C.: Abnormal sperm parameters in humans are indicative of an abortive apoptotic mechanism linked to the Fas-mediated pathway. Exp. Cell. Res., 1999, 251: 350–355.

    PubMed  Article  CAS  Google Scholar 

  25. 25.

    SUN J.G., JURISICOVA A., CASPER R.F.: Detection of deoxyribonucleic acid fragmentation in human sperm: correlation with fertilization in vitro. Biol. Reprod., 1997, 56: 602–607.

    PubMed  Article  CAS  Google Scholar 

  26. 26.

    THOMAS C.A., GARNER D.L.: Post-thaw bovine spermatozoal quality estimated from fresh samples. J. Androl., 1994, 15: 305–340.

    Google Scholar 

  27. 27.

    TSAI Y.L., LIU J., GARCIA J.E., KATZ E., COMPTON G., BARAMKI T.A.: Establishment of an optimal hypo-osmotic swelling test by examining single spermatozoa in four different hypo-osmotic solutions. Hum. Reprod., 1997, 12: 1111–1113.

    PubMed  Article  CAS  Google Scholar 

  28. 28.

    VAN DEN SAFFELE J., VERMEULEN L., SCHOONJANS F., COMHAIRE F.H.: Evaluation of the hypo-osmotic swelling test in relation with advanced methods of semen analysis. Andrologia, 1992, 24: 213–217.

    PubMed  Google Scholar 

  29. 29.

    VENDRELL F.J., RUBIO C., TARIN J.J.: The heparin-glutathion test: an alternative to the hypo-osmotic swelling test to select viable sperm for intracytoplasmic sperm injection. Fertil. Steril., 1998, 70: 1156–1161.

    PubMed  Article  CAS  Google Scholar 

  30. 30.

    WHO Laboratory Manual for the Examination of Human Semen and Sperm-Cervical Mucus Interaction. London, Cambridge University Press, 4th edition, 1999.

  31. 31.

    ZAMBONI L.: The ultrastructural pathology of the spermatozoon as a cause of infertility: the role of electron microscopy in the evaluation of semen quality. Fertil. Steril., 1987, 48: 711–734.

    PubMed  CAS  Google Scholar 

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Correspondence to T. Forges.

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Communication aux 3èmes Journées Nationales Assurance de Qualité en Biologie de Reproduction, 26 & 27 October 2000, Nancy.

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Forges, T., Monnier-Barbarino, P. & Foliguet, B. La vitalité des spermatozoïdes. Androl. 11, 45–55 (2001). https://doi.org/10.1007/BF03034509

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Mots clés

  • Vitalité
  • Spermatozoïde
  • éosine
  • HOST
  • ICSI
  • Apoptose

Key words

  • Viability
  • Spermatozoa
  • Eosin-Y
  • HOST
  • ICSI
  • Apoptosis
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