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01-01-2014 | Review Article

Observation of spermatozoa by a high-magnification microscope

Authors: Akira Komiya, Akihiko Watanabe, Tomonori Kato, Yoko Kawauchi, Hideki Fuse

Published in: Reproductive Medicine and Biology | Issue 1/2014

Abstract

Semen analyses are the primary tool for evaluating male infertility, as semen parameters are useful for predicting potential fertility. In the field of assisted reproductive technology (ART), the single best motile spermatozoon should be selected, especially when performing intracytoplasmic sperm injection (ICSI). In this context, the motile sperm organelle morphology examination (MSOME) was developed as a method of assessing the detailed morphology of motile spermatozoa in real time at a magnification of up to 6,300× on a video system. The use of ICSI with MSOME-selected sperm is called intracytoplasmic morphologically selected sperm injection (IMSI). IMSI improves the outcomes of ICSI. MSOME can be also applied to evaluate male infertility. Among MSOME parameters, the presence of sperm nuclear vacuoles is the most important finding. Large sperm nuclear vacuoles (LNV) are related not only to poor ART outcomes, but also to poor semen quality and sperm DNA damage, such as DNA fragmentation and chromatin condensation failure. It has been suggested that sperm head vacuoles are produced at earlier stages of sperm maturation. It is possible that the number of LNV can be decreased by surgical or medical treatment for male infertility. Therefore, the level of LNV has the potential to be used as an alternative parameter of semen quality and a new tool for evaluating the therapeutic effects of treatment in male patients with infertility.

Palermo G, Joris H, Devroey P, Van Steirteghem A. Pregnancies after intracytoplasmic injection of single spermatozoon into an oocyte. Lancet. 1992;340:17–8.PubMedCrossRef

Jakab A, Sakkas D, Delpiano E, Cayli S, Kovanci E, Ward D, et al. Intracytoplasmic sperm injection: a novel selection method for sperm with normal frequency of chromosomal aneuploidies. Fertil Steril. 2005;84:1665–73.PubMedCrossRef

Sakkas D. Novel technologies for selecting the best sperm for in vitro fertilization and intracytoplasmic sperm injection. Fertil Steril. 2013;99:1023–9.PubMedCrossRef

Worrilow K, Eid S, Woodhouse D, Perloe M, Smith S, Witmyer J, et al. Use of hyaluronan in the selection of sperm for intracytoplasmic sperm injection (ICSI): significant improvement in clinical outcomes—multicenter, double-blinded and randomized controlled trial. Hum Reprod. 2013;28:306–14.PubMedCrossRef

Rogers BJ, Bentwood BJ, Van Campen H, Helmbrecht G, Soderdahl D, Hale RW. Sperm morphology assessment as an indicator of human fertilizing capacity. J Androl. 1983;4:119–25.

Ombelet W, Bosmans E, Janssen M, Cox A, Vlasselaer J, Gyselaers W, et al. Semen parameters in a fertile versus subfertile population: a need for change in the interpretation of semen testing. Hum Reprod. 1997;12:987–93.PubMedCrossRef

Zinaman M, Brown C, Selevan S, Clegg E. Semen quality and human fertility: a prospective study with healthy couples. J Androl. 2000;21:145–53.PubMed

Menkveld R, Wong W, Lombard Y, Wetzels CJ, Thomas AMM, Merkus CMG, Steegers-Theunissen HMW. Semen parameters, including WHO and strict criteria morphology, in a fertile and subfertile population: an effort towards standarization of in vivo thresholds. Hum Reprod. 2001;16:1165–71.PubMedCrossRef

Guzick DS, Overstreet JW, Factor-Litvak P, Brazil CK, Nakajima ST, Coutifaris C, et al. Sperm morphology, motility, and concentration in fertile and infertile men. N Engl J Med. 2001;345:1388–93.PubMedCrossRef

10.

Jedrzejczak P, Taszarek-Hauke G, Hauke J, Pawelczyk L, Duleba AJ. Prediction of spontaneous conception based on semen parameters. Int J Androl. 2008;31:499–507.PubMedCrossRef

11.

Akashi T, Watanabe A, Komiya A, Fuse H. Evaluation of the sperm motility analyzer system (SMAS) for the assessment of sperm quality in infertile men. Syst Biol Reprod Med. 2010;56:473–7.PubMedCrossRef

12.

WHO. Semen analysis. In: Organization WH, editor. WHO laboratory manual for the examination and processing of human semen. 5th ed. Switzerland: WHO; 2010. p. 7–115.

13.

Fetterolf PM, Rogers BJ. Prediction of human sperm penetrating ability using computerized motion parameters. Mol Reprod Dev. 1990;27:326–31.PubMedCrossRef

14.

Hirano Y, Shibahara H, Obara H, Suzuki T, Takamizawa S, Yamaguchi C, et al. Relationships between sperm motility characteristics assessed by the computer-aided sperm analysis (CASA) and fertilization rates in vitro. J Assist Reprod Genet. 2001;18:213–8.PubMedCrossRef

15.

Parinaud J, Richoilley G, Moutaffian H, Vieitez G, Mieusset R. Are the characteristics of spermatozoa in the insemination medium useful for predicting in vitro fertilization results? Int J Androl. 1996;19:103–8.PubMedCrossRef

16.

Seiringer M, Maurer M, Shebl O, Dreier K, Tews G, Ziehr S, et al. Efficacy of a sperm-selection chamber in terms of morphology, aneuploidy and DNA packaging. Reprod Biomed Online. 2013. doi:10.1016/j.rbmo.2013.03.013 (Epub ahead of print, in press).

17.

Mashiach R, Fisch B, Eltes F, Tadir Y, Ovadia J, Bartoov B. The relationship between sperm ultrastructural features and fertilizing capacity in vitro. Fertil Steril. 1992;57:1052–7.PubMed

18.

Bartoov B, Eltes F, Pansky M, Langzam J, Reichart M, Soffer Y. Improved diagnosis of male fertility potential via a combination of quantitative ultramorphology and routine semen analyses. Hum Reprod. 1994;9:2069–75.PubMed

19.

Bartoov B, Eltes F, Soffer Y, Ron-El R, Langzam J, Ledcrman H. Ultramorphological characteristics of human sperm cells. In: Mashiach S, Ben-Rafael Z, Laufer N, Schenkler JG, editors. Advances in in vitro fertilization and assisted reproduction technologies. New York: Plenum Publishers; 1990.

20.

Berkovitz A, Eltes F, Soffer Y, Zabludovsky N, Beyth Y, Farhi J, et al. ART success and in vivo sperm cell selection depend on the ultramorphological status of spermatozoa. Andrologia. 1999;31:1–8.PubMed

21.

Bartoov B, Berkovitz A, Eltes F. Selection of spermatozoa with normal nuclei to improve the pregnancy rate with intracytoplasmic sperm injection. N Engl J Med. 2001;345:1067–8.PubMedCrossRef

22.

Centonze Frohlich V. Phase contrast and differential interference contrast (DIC) microscopy. J Vis Exp. 2008.

23.

Bartoov B, Berkovitz A, Eltes F, Kogosowski A, Menezo Y, Barak Y. Real-time fine morphology of motile human sperm cells is associated with IVF-ICSI outcome. J Androl. 2002;23:1–8.PubMed

24.

Glezerman M, Bartoov B. Semen analysis. In: Insler V, Lunenfeld B, editors. Infertility: male and female. Edinburgh: Churchill Livingstone; 1993.

25.

Bartoov B, Berkovitz A, Eltes F, Kogosovsky A, Yagoda A, Lederman H, et al. Pregnancy rates are higher with intracytoplasmic morphologically selected sperm injection than with conventional intracytoplasmic injection. Fertil Steril. 2003;80:1413–9.PubMedCrossRef

26.

Berkovitz A, Eltes F, Yaari S, Katz N, Barr I, Fishman A, et al. The morphological normalcy of the sperm nucleus and pregnancy rate of intracytoplasmic injection with morphologically selected sperm. Hum Reprod. 2005;20:185–90.PubMedCrossRef

27.

Berkovitz A, Eltes F, Ellenbogen A, Peer S, Feldberg D, Bartoov B. Does the presence of nuclear vacuoles in human sperm selected for ICSI affect pregnancy outcome? Hum Reprod. 2006;21:1787–90.PubMedCrossRef

28.

de Almeida Ferreira Braga DP, Setti AS, Figueira RC, Nichi M, Martinhago CD, Iaconelli Jr A. Sperm organelle morphologic abnormalities: contributing factors and effects on intracytoplasmic sperm injection cycles outcomes. Urology. 2011;78:786–91.PubMedCrossRef

29.

Balaban B, Yakin K, Alatas C, Oktem O, Isiklar A, Urman B. Clinical outcome of intracytoplasmic injection of spermatozoa morphologically selected under high magnification: a prospective randomized study. Reprod Biomed Online. 2011.

30.

Setti AS, Figueira RC, Paes de Almeida Ferreira Braga D, de Cassia Savio Figueira R, Iaconelli Jr A, Borges Jr E. Intracytoplasmic sperm injection outcome versus intracytoplasmic morphologically selected sperm injection outcome: a meta-analysis. Reprod Biomed Online. 2010;21:450–5.CrossRef

31.

Gonzalez-Ortega C, Cancino-Villarreal P, Perez-Torres A, Vargas-Maciel MA, Martinez-Garza SG, Perez-Pena E, et al. Intracytoplasmic morphologically selected sperm injection (IMSI) vs. intracytoplasmic sperm injection (ICSI) in patients with repeated ICSI failure. Ginecol Obstet Mex. 2010;78:652–9.PubMed

32.

Mauri AL, Petersen CG, Oliveira JB, Massaro FC, Baruffi RL, Franco JG. Comparison of day 2 embryo quality after conventional ICSI versus intracytoplasmic morphologically selected sperm injection (IMSI) using sibling oocytes. Eur J Obstet Gynecol Reprod Biol. 2010;150:42–6.PubMedCrossRef

33.

Tanaka A, Nagayoshi M, Tanaka I, Kusunoki H. Human sperm head vacuoles are physiological structures formed during the sperm development and maturation process. Fertil Steril. 2012;98:315–20.PubMedCrossRef

34.

Silva LF, Oliveira JB, Petersen CG, Mauri AL, Massaro FC, Cavagna M, et al. The effects of male age on sperm analysis by motile sperm organelle morphology examination (MSOME). Reprod Biol Endocrinol. 2012;10:19.PubMedCentralPubMedCrossRef

35.

Jacquez JA, Biesele JJ. Nuclear vacuoles in spermatids of C-57 black mice after injection of beta-1 naphthylalanine. Anat Rec. 1951;109:699–705.PubMedCrossRef

36.

Billard R. Spermiogenesis in the rainbow trout (Salmo gairdneri). An ultrastructural study. Cell Tissue Res. 1983;233:265–84.PubMedCrossRef

37.

Czaker R. On the origin of nuclear vacuoles in spermatozoa a fine structural and cytochemical study in mice. Andrologia. 1985;17:547–57.PubMedCrossRef

38.

Bonet S, Briz M, Fradera A. Ultrastructural abnormalities of boar spermatozoa. Theriogenology. 1993;40:383–96.PubMedCrossRef

39.

Thundathil J, Palasz AT, Barth AD, Mapletoft RJ. Fertilization characteristics and in vitro embryo production with bovine sperm containing multiple nuclear vacuoles. Mol Reprod Dev. 1998;50:328–33.PubMedCrossRef

40.

Parkinson TJ. Nuclear vacuolation as a cause of sterility in an Angus bull. Vet J. 2000;159:207–10.PubMedCrossRef

41.

Revay T, Nagy S, Kopp C, Flyckt A, Rens W, Rath D, et al. Macrocephaly in bull spermatozoa is associated with nuclear vacuoles, diploidy and alteration of chromatin condensation. Cytogenet Genome Res. 2009;126:202–9.PubMedCrossRef

42.

Komiya A, Watanabe A, Kawauchi Y, Fuse H. Sperm with large nuclear vacuoles and semen quality in the evaluation of male infertility. Syst Biol Reprod Med. 2013;59:13–20.PubMedCrossRef

43.

Falagario D, Brucculeri AM, Depalo R, Trerotoli P, Cittadini E, Ruvolo G. Sperm head vacuolization affects clinical outcome in ICSI cycle. A proposal of a cut-off value. J Assist Reprod Genet. 2012;29:1281–7.PubMedCrossRef

44.

Vanderzwalmen P, Hiemer A, Rubner P, Bach M, Neyer A, Stecher A, et al. Blastocyst development after sperm selection at high magnification is associated with size and number of nuclear vacuoles. Reprod Biomed Online. 2008;17:617–27.PubMedCrossRef

45.

Boitrelle F, Albert M, Theillac C, Ferfouri F, Bergere M, Vialard F, et al. Cryopreservation of human spermatozoa decreases the number of motile normal spermatozoa, induces nuclear vacuolization and chromatin decondensation. J Androl. 2012;33:1371–8.PubMedCrossRef

46.

Garolla A, Fortini D, Menegazzo M, De Toni L, Nicoletti V, Moretti A, et al. High-power microscopy for selecting spermatozoa for ICSI by physiological status. Reprod Biomed Online. 2008;17:610–6.PubMedCrossRef

47.

Boitrelle F, Ferfouri F, Petit JM, Segretain D, Tourain C, Bergere M, et al. Large human sperm vacuoles observed in motile spermatozoa under high magnification: nuclear thumbprints linked to failure of chromatin condensation. Hum Reprod. 2011;26:1650–8.PubMedCrossRef

48.

Watanabe S, Tanaka A, Fujii S, Mizunuma H, Fukui A, Fukuhara R, et al. An investigation of the potential effect of vacuoles in human sperm on DNA damage using a chromosome assay and the TUNEL assay. Hum Reprod. 2011;26:978–86.PubMedCrossRef

49.

Perdrix A, Saidi R, Menard JF, Gruel E, Milazzo JP, Mace B, et al. Relationship between conventional sperm parameters and motile sperm organelle morphology examination (MSOME). Int J Androl. 2012;35:491–8.PubMedCrossRef

50.

Oliveira JB, Petersen CG, Massaro FC, Baruffi RL, Mauri AL, Silva LF, et al. Motile sperm organelle morphology examination (MSOME): intervariation study of normal sperm and sperm with large nuclear vacuoles. Reprod Biol Endocrinol. 2010;8:56.PubMedCentralPubMedCrossRef

51.

Sermondade N, Vialard F, Berge`re M, Hammoud I, Cavelot P, Selva J, et al. Evaluation de l’apport de la me′ thode d’observation des spermatozo¨ıdes a` fort grossissement en ICSI. Andrologie. 2007;17:212–21.CrossRef

52.

De Kretser DM. Ultrastructural features of human spermiogenesis. Zeitschrift fur Zellforschung und mikroskopische Anatomie. 1969;98:477–505.PubMedCrossRef

53.

Sousa M, Barros A, Takahashi K, Oliveira C, Silva J, Tesarik J. Clinical efficacy of spermatid conception: analysis using a new spermatid classification scheme. Hum Reprod. 1999;14:1279–86.PubMedCrossRef

54.

Toshimori K, Ito HC. Human sperm ultrastructures and fertility. J Mamm Ova Res. 2008;25:232–9.CrossRef

55.

Toshimori K. Dynamics of the mammalian sperm head: modifications and maturation events from spermatogenesis to egg activation. Adv Anat Embryol Cell Biol. 2009;204:5–94.PubMed

56.

Perdrix A, Travers A, Chelli MH, Escalier D, Do Rego JL, Milazzo JP, et al. Assessment of acrosome and nuclear abnormalities in human spermatozoa with large vacuoles. Hum Reprod. 2011;26:47–58.PubMedCrossRef

57.

Chen SS, Huang WJ, Chang LS, Wei YH. 8-Hydroxy-2′-deoxyguanosine in leukocyte DNA of spermatic vein as a biomarker of oxidative stress in patients with varicocele. J Urol. 2004;172:1418–21.PubMedCrossRef

58.

Agarwal A, Said TM. Oxidative stress, DNA damage and apoptosis in male infertility: a clinical approach. BJU Int. 2005;95:503–7.PubMedCrossRef

59.

Smith R, Kaune H, Parodi D, Madariaga M, Rios R, Morales I, et al. Increased sperm DNA damage in patients with varicocele: relationship with seminal oxidative stress. Hum Reprod. 2006;21:986–93.PubMedCrossRef

60.

Aitken RJ, Wingate JK, De Iuliis GN, Koppers AJ, McLaughlin EA. Cis-unsaturated fatty acids stimulate reactive oxygen species generation and lipid peroxidation in human spermatozoa. J Clin Endocrinol Metab. 2006;91:4154–63.PubMedCrossRef

61.

Ishikawa T, Fujioka H, Ishimura T, Takenaka A, Fujisawa M. Increased testicular 8-hydroxy-2′-deoxyguanosine in patients with varicocele. BJU Int. 2007;100:863–6.PubMedCrossRef

62.

Dada R, Shamsi MB, Venkatesh S, Gupta NP, Kumar R. Attenuation of oxidative stress and DNA damage in varicocelectomy: implications in infertility management. Indian J Med Res. 2010;132:728–30.PubMed

63.

O’Brien J, Zini A. Sperm DNA integrity and male infertility. Urology. 2005;65:16–22.PubMedCrossRef

64.

Zini A, Blumenfeld A, Libman J, Willis J. Beneficial effect of microsurgical varicocelectomy on human sperm DNA integrity. Hum Reprod. 2005;20:1018–21.PubMedCrossRef

65.

Zini A, Libman J. Sperm DNA damage: clinical significance in the era of assisted reproduction. CMAJ. 2006;175:495–500.PubMedCentralPubMedCrossRef

66.

Fuse H, Akashi T, Mizuno I, Nozaki T, Watanabe A. Postoperative changes of sperm chromatin heterogeneity, using acridine orange staining, in varicocele patients. Arch Androl. 2006;52:223–6.PubMedCrossRef

67.

Medicine PCoASfR. The clinical utility of sperm DNA integrity testing. Fertil Steril. 2008;90:S178–80.

68.

Papachristou F, Simopoulou M, Touloupidis S, Tsalikidis C, Sofikitis N, Lialiaris T. DNA damage and chromosomal aberrations in various types of male factor infertility. Fertil Steril. 2008;90:1774–81.PubMedCrossRef

69.

Moskovtsev SI, Lecker I, Mullen JB, Jarvi K, Willis J, White J, et al. Cause-specific treatment in patients with high sperm DNA damage resulted in significant DNA improvement. Syst Biol Reprod Med. 2009;55:109–15.PubMedCrossRef

70.

Menezo Y, Dale B, Cohen M. DNA damage and repair in human oocytes and embryos: a review. Zygote. 2010;18:357–65.PubMedCrossRef

71.

Moskovtsev SI, Mullen JB, Lecker I, Jarvi K, White J, Roberts M, et al. Frequency and severity of sperm DNA damage in patients with confirmed cases of male infertility of different aetiologies. Reprod Biomed Online. 2010;20:759–63.PubMedCrossRef

72.

Sakkas D, Alvarez JG. Sperm DNA fragmentation: mechanisms of origin, impact on reproductive outcome, and analysis. Fertil Steril. 2010;93:1027–36.PubMedCrossRef

73.

Oliveira JB, Massaro FC, Baruffi RL, Mauri AL, Petersen CG, Silva LF, et al. Correlation between semen analysis by motile sperm organelle morphology examination and sperm DNA damage. Fertil Steril. 2010;94:1937–40.PubMedCrossRef

74.

Skowronek F, Casanova G, Alciaturi J, Capurro A, Cantu L, Montes JM, et al. DNA sperm damage correlates with nuclear ultrastructural sperm defects in teratozoospermic men. Andrologia. 2012;44:59–65.PubMedCrossRef

75.

Gopalkrishnan K, Padwal V, Meherji PK, Gokral JS, Shah R, Juneja HS. Poor quality of sperm as it affects repeated early pregnancy loss. Arch Androl. 2000;45:111–7.PubMedCrossRef

76.

Franco JG Jr, Mauri AL, Petersen CG, Massaro FC, Silva LF, Felipe V, et al. Large nuclear vacuoles are indicative of abnormal chromatin packaging in human spermatozoa. Int J Androl. 2012;35:46–51.PubMedCrossRef

77.

Cassuto NG, Hazout A, Hammoud I, Balet R, Bouret D, Barak Y, et al. Correlation between DNA defect and sperm-head morphology. Reprod Biomed Online. 2012;24:211–8.PubMedCrossRef

78.

Utsuno H, Oka K, Yamamoto A, Shiozawa T. Evaluation of sperm head shape at high magnification revealed correlation of sperm DNA fragmentation with aberrant head ellipticity and angularity. Fertil Steril. 2013.

79.

Tanii I, Aradate T, Matsuda K, Komiya A, Fuse H. PACAP-mediated sperm–cumulus cell interaction promotes fertilization. Reproduction. 2011;141:163–71.PubMedCrossRef

80.

Monqaut AL, Zavaleta C, Lopez G, Lafuente R, Brassesco M. Use of high-magnification microscopy for the assessment of sperm recovered after two different sperm processing methods. Fertil Steril. 2011;95:277–80.PubMedCrossRef

81.

Montjean D, Belloc S, Benkhalifa M, Dalleac A, Menezo Y. Sperm vacuoles are linked to capacitation and acrosomal status. Hum Reprod. 2012;27:2927–32.PubMedCrossRef

82.

Kacem O, Sifer C, Barraud-Lange V, Ducot B, De Ziegler D, Poirot C, et al. Sperm nuclear vacuoles, as assessed by motile sperm organellar morphological examination, are mostly of acrosomal origin. Reprod Biomed Online. 2010;20:132–7.PubMedCrossRef

83.

Neyer A, Vanderzwalmen P, Bach M, Stecher A, Spitzer D, Zech N. Sperm head vacuoles are not affected by in vitro conditions, as analysed by a system of sperm-microcapture channels. Reprod Biomed Online. 2012.

84.

Furuya Y, Akashi T, Fuse H. Effect of Bu-zhong-yi-qi-tang on seminal plasma cytokine levels in patients with idiopathic male infertility. Arch Androl. 2004;50:11–4.PubMedCrossRef

85.

Furuya Y, Akashi T, Fuse H. Treatment of traditional Chinese medicine for idiopathic male infertility. Hinyokika Kiyo. 2004;50:545–8.PubMed

86.

Komiya A, Watanabe A, Fuse H. Herbal medicine in Japan. J Mens Health. 2011;8:S15–8.CrossRef

87.

Komiya A, Watanabe A, Kawauchi Y, Fuse H. Observation of changes in sperm nuclear vacuoles among patients with male infertility. Andrology. 2013;59(Suppl. 1):74.

Title: Observation of spermatozoa by a high-magnification microscope
Authors: Akira Komiya
Akihiko Watanabe
Tomonori Kato
Yoko Kawauchi
Hideki Fuse
Publication date: 01-01-2014
Publisher: Springer Japan
Published in: Reproductive Medicine and Biology / Issue 1/2014
Print ISSN: 1445-5781
Electronic ISSN: 1447-0578
DOI: https://doi.org/10.1007/s12522-013-0164-y

Keynote webinar | Spotlight on medication adherence

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Observation of spermatozoa by a high-magnification microscope

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