ICSI, Male Pronuclear Remodeling and Cell Cycle Checkpoints

  • Laura Hewitson
  • Calvin R. Simerly
  • Gerald Schatten
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 518)


Intracytoplasmic sperm injection (ICSI; Palermo et al., 1992; Van Steirteghem et al., 1993) has heralded an era of enormous improvements in treating male infertility; it has resulted in the births of thousands of babies. In fact its success has led some clinicians to claim that male infertility is now “cured”, while others raise concerns over the global application of ICSI for all cases of in vitro fertilization (Tucker et al., 1995). These concerns may be, in part, based on the lack of supporting pre-clinical studies in relevant animal models (Ng et al., 1995; Yanagimachi et al, 1995). Others are worried about the possible long-term effects of ICSI on the offspring (Hamberger et al., 1998]; Moutel et al., 1999). While the current view of ICSI is one of cautious optimism, the development of a preclinical, non-human primate model has been established to address the safety of ICSI and other methods of assisted reproduction (Hewitson et al., 1998; Hewitson et al., 1999; Hewitson et al., 2001). Using advanced fluorescent imaging techniques, the cytoskeletal events that occur during rhesus fertilization have been examined after both in vitro fertilization (IVF) and ICSI. It was found that rhesus monkeys share many similarities with humans in terms of cytoskeletal and chromatin dynamics after fertilization by IVF (Simerly et al., 1995; Wu et al., 1996). However, rhesus oocytes fertilized by ICSI resulted in abnormal nuclear remodeling leading to asynchronous chromatin decondensation in the apical region of the sperm head, delaying the onset of DNA synthesis. The persistence of the acrosome and perinuclear theca on the apex of sperm introduced into the oocyte by ICSI may constrict the DNA in this region (Hewitson et al., 1999; Ramalho-Santos et al., 2000a). Despite these differentces, normal rhesus ICSI embryos have been produced and, after transfer to surrogate females, have resulted in several births (Hewitson et al., 1999; Hewitson et al., 2001)


Rhesus Monkey Assisted Reproduction Intracytoplasmic Sperm Injection Meiotic Spindle Fertilization Failure 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Asch, R., Simerly, C., Ord, T., Ord, V.A., and Schatten, G., 1995, The stages at which human fertilization arrests: microtubule and chromosome configurations in inseminated oocytes which fail to complete fertilization and development in humans. Hum Reprod. 10:1897–1906.PubMedGoogle Scholar
  2. Bonduelle, M., Joris, H., Hofmans, K., Liebaers, I., and A. Van Steirteghem, 1998a, Mental development of 201 ICSI children at 2 years of age. Lancet. 351:1553.Google Scholar
  3. Bonduelle, M., Aytoz, A., and Van Assche, E., 1998b, Incidence of chromosomal aberrations in children born after assisted reproduction through intracytoplasmic sperm injection. Hum Reprod. 13:781–782.PubMedCrossRefGoogle Scholar
  4. Bourne, H., Richings, N., Harari, O., Watkins, W., Speirs, A.L., Johnston, W.I., and Baker, H.W., 1995, The use of intracytoplasmic sperm injection for the treatment of severe and extreme male infertility. Reprod Fertil Dev. 7:237–245.PubMedCrossRefGoogle Scholar
  5. Bowen, J.R., Gibson, F.L., Leslie, G.I. and Saunders, D.M., 1998, Medical and developmental outcome at 1 year for children conceived by intracytoplasmic sperm injection. Lancet. 351:1529–1534.PubMedCrossRefGoogle Scholar
  6. Chan, A.W.S., Luetjens, C.M., Dominko, T., Ramalho-Santos, J., Hewitson, L., Simerly, C., and Schatten, G., 2000, TransgenlCSI: Foreign DNA transmission by intracytoplasmic sperm injection: Injection of sperm bound with exogenous DNA results in embryonic GFP expression and live rhesus births. Mol Hum Reprod. 6:26–33.Google Scholar
  7. Compton, D. A., and Cleveland, D.W., 1994, NuMA, a nuclear protein involved in mitosis and nuclear reformation. Curr Opin Cell Biol. 6:343–346.PubMedCrossRefGoogle Scholar
  8. Conner, S., Leaf, D., and Wessel, G., 1997, Members of the SNARE hypothesis are associated with cortical granule exocytosis in the sea urchin egg. Mol Reprod Dev. 48:106–118.PubMedCrossRefGoogle Scholar
  9. Cummins, J.M., and Jequier, A.M., 1995, Concerns and recommendations for intracytoplasmic sperm injection (ICSI) treatment. Hum Reprod. 10, Suppl 1:138–143.PubMedCrossRefGoogle Scholar
  10. Devroey, P., Silber, S., Nagy, Z., Liu, J., Tournaye, H., Joris, H., Verheyen, G., and Van Steirteghem, A., 1995, Ongoing pregnancies and birth after intracytoplasmic sperm injection with frozen-thawed epididymal spermatozoa. Hum Reprod. 10:903–906.PubMedCrossRefGoogle Scholar
  11. Dowsing, A.T., Yong, EX., Clark, M., McLachlan, R.I., de Kretser, D.M., and Trounson, A.O., 1999, Linkage between male infertility and trinucleotide repeat expansion in the androgen-reccptor gene. Lancet. 354:640–643.PubMedCrossRefGoogle Scholar
  12. Dumoulin, J., Coonen, E., Bras, M., Bergers-Janssen, J., Ignoul-Vanvuchelen, R., van Wissen, L., Geraedts, J., and Evers, J., 2001, Embryo development and chromosomal anomalies after ICSI: effect of the injection procedure. Hum Reprod. 16:306–312.PubMedCrossRefGoogle Scholar
  13. Fishel, S., Green, S., and Bishop, M., 1995, Pregnancy after intracytoplasmic injection of a spermatid. Lancet. 345:1641–1642.PubMedCrossRefGoogle Scholar
  14. Hamberger, L., Lundin, K., Sjogren, A., and Soderlund, B., 1998, Indications for intracytoplasmic sperm injection. Hum Reprod. 13,Suppl 1:128–133.CrossRefGoogle Scholar
  15. Harari, O., Bourne, H., Baker, G., Gronow, M., and Johnston I., 1995, High fertilization rate with intracytoplasmic sperm injection in mosaic Klinefelter’s syndrome. Fertil Steril. 63:182–184.PubMedGoogle Scholar
  16. Hargreave, T.B., 2000, Genetics and male infertility. Curr Opin Obst Gyn. 12:207–219.CrossRefGoogle Scholar
  17. Hewitson, L., Simerly, C., Tengowski, M., Sutovsky, P., Navara, C., Haavisto, A.J., and Schatten, G., 1996, Microtubule and chromatin configurations during rhesus intracytoplasmic sperm injection: successes and failures. Biol Reprod. 55:271–280.PubMedCrossRefGoogle Scholar
  18. Hewitson, L., Takahashi, D., Dominko, T., Simerly, C., and Schatten, G., 1998, Fertilization and embryo development to blastocysts after intracytoplasmic sperm injection in the rhesus monkey. Hum Reprod.l3:3449–3455.CrossRefGoogle Scholar
  19. Hewitson, L., Dominko, T., Takahashi, D., Martinovich, C., Ramalho-Santos, J., Sutovsky, P., Fanton, J., Jacob, D., Monteith, D., Neuringer, M., Battaglia, D., Simerly C., and Schatten, G., 1999, Unique checkpoints during the first cell cycle of fertilization after intracytoplasmic sperm injection in rhesus monkeys. Nat Med. 5:431–433.PubMedCrossRefGoogle Scholar
  20. Hewitson, L., Martinovich, C., Simerly, C., Takahashi, D., and Schatten, G., 2001, Intracytoplasmic injection of rhesus testicular sperm (TESE-ICSI) or elongated spermatids (ELSI) gives rise to healthy offspring. Fertil Steril. 77:794–801.CrossRefGoogle Scholar
  21. Kamischke, A., Gromoll, J., Simoni, M., Behre, H.M., and Nieschlag, E., 1999, Transmission of a Y chromosomal deletion involving the deleted in azoospermia (DAZ) and chromodomain (CDY1) genes from father to son through intracytoplasmic sperm injection: case report. Hum Reprod. 14:2320–2322.PubMedCrossRefGoogle Scholar
  22. Keefer, C.L., Younis, A.I., and Brackett, B.G., 1990, Cleavage development of bovine oocytcs fertilized by sperm injection. Mol Reprod Dev. 25:281–285.PubMedCrossRefGoogle Scholar
  23. Kent-First, M. G., Kol, S., Muallem, A., Ofir, R., Manor, D., Blazer, S., First, N., and Itskovitz-Eldor, J., 1996, The incidence and possible relevance of Y-linked microdeletions in babies bora after intracytoplasmic sperm injection and their infertile fathers. Mol Hum Reprod. 2:943–950.PubMedCrossRefGoogle Scholar
  24. Kent-First,M., 2000, The critical and expanding role of genetics in assisted reproduction. Prenat Diagn. 20:536–551.PubMedCrossRefGoogle Scholar
  25. Kim, E.D., Bischoff, F.Z., Lipshultz, L.I., and Lamb, D.J., 1998, Genetic concerns for the subfertile male in the era of ICSI. Prenat Diagn. 18:1349–1365.PubMedCrossRefGoogle Scholar
  26. Kim, N.H., Jun, S.H., Do, J.T., Uhm, S.J., Lee, H.T., Chung, K.S., 1999, Intracytoplasmic injection of porcine, bovine, mouse, or human spermatozoon into porcine oocytes. Molec Reprod Dev. 53:84–91.PubMedCrossRefGoogle Scholar
  27. Kupker, W., Schwinger, E., Hiort, O., Ludwig, M., Nikolettos, N., Schlegel, P.N., and Diedrich K, 1999, Genetics of male subfertility: consequences for the clinical work-up. Hum Reprod. 14, Suppl 1:24–37.PubMedCrossRefGoogle Scholar
  28. Lamb, D.J., and Lipshultz L.I., 2000, Male infertility: recent advances and a look towards the future. Curr Opin Urol. 10:359–362.Google Scholar
  29. Liu, J., Nagy, Z., Joris, H., Tournaye, H., Devroey, P., and Van Steirteghem, A., 1995, Successful fertilization and establishment of pregnancies after intracytoplasmic sperm injection in patients with globozoospermia. Hum Reprod. 10:626–629.PubMedGoogle Scholar
  30. Luetjens, C.M., Payne, C., and Schatten, G., 1999, Non-random chromosome positioning in human sperm and sex chromosome anomalies following intracytoplasmic sperm injection. Lancet. 353:1240.PubMedCrossRefGoogle Scholar
  31. Meschede, D., Lemcke, B., Behre, H.M., De Geyter, C., Nieschlag, E., and Horst, J., 2000, Clustering of male infertility in the families of couples treated with intracytoplasmic sperm injection. Hum Reprod. 15:1604–1608.PubMedCrossRefGoogle Scholar
  32. Moutel, G., Leroux, N., and Herve, C., 1999, Keeping an eye on ICSI. Nature Med. 5:593.PubMedCrossRefGoogle Scholar
  33. Ng, S.C., Liow, S.L., Ahmadi, A., Yong, EX., Bongso, A., and Ratnam, S.S., 1995, Intracytoplasmic sperm injection—is there a need for an animal model, especially in assessing the genetic risks involved?. Hum Reprod. 10:2523–2525.PubMedGoogle Scholar
  34. Nudell, D., Castillo, M., Turek, P.J., and Pera, R.R., 2000, Increased frequency of mutations in DNA from infertile men with meiotic arrest. Hum Reprod. 15:1289–1294.PubMedCrossRefGoogle Scholar
  35. Palermo, G., Joris, H., Devroey, P., and Van Steirteghem, A., 1992, Pregnancies after intracytoplasmic sperm injection of single spermatozoon into an oocyte. Hum Reprod. 340:17–18.Google Scholar
  36. Pedcrsen, R.A., 2001, Sperm and mammalian polarity. Nature. 409:473–474.CrossRefGoogle Scholar
  37. Perry, A.C., Wakayama, T., Kishikawa, H., Kasai, T., Okabe, M., Toyoda, Y., and Yanagimachi, R., 1999, Mammalian transgenesis by intracytoplasmic sperm injection. Science. 284:1180–1183.PubMedCrossRefGoogle Scholar
  38. Ramalho-Santos, J., Sutovsky, P., Simerly, C.R., Oko, R., Wessel, G.M., Hewitson,L., and Schatten, G., 2000a, ICSI choreography: Fate of sperm structures after monospermic ICSI and first cell cycle implications. Hum Reprod. 15:2610–2620.PubMedCrossRefGoogle Scholar
  39. Ramalho-Santos, J., Moreno, R.D., Sutovsky, P., Chan, A.W.-S., Hewitson, L., Wessel, G., Simerly, C., and Schatten, G., 2000b, SNAREs in mammalian sperm mediate membrane fusion events during fertilization. Dev Biol. 223:54–69.PubMedCrossRefGoogle Scholar
  40. Rawe, V.Y, Olmedo, S.B., Nodar, F.N., Doncel, G.D., Acosta, A.A., and Vitullo, A.D., 2000, Cytoskeletal organization defects and abortive activation in human oocytes after IVF and ICSI failure. Mol Hum Reprod. 6:510–516.PubMedCrossRefGoogle Scholar
  41. Rybouchkin, A.V., Van der Straeten, F., Quatacker, J., De Sutter, P., and Dhont, M., 1997, Fertilization and pregnancy after assisted oocyte activation and intracytoplasmic sperm injection in a case of round-headed sperm associated with deficient oocyte activation capacity. Fertil Steril. 68:1144–1147PubMedCrossRefGoogle Scholar
  42. Schatten, G., Simerly, C., and Schatten, H, 1985, Microtubule configurations during fertilization, mitosis, and early development in the mouse and the requirement for egg microtubule-mediated motility during mammalian fertilization. Proc Natl Acad Sci. 82:4152–4156.PubMedCrossRefGoogle Scholar
  43. Schatten, G., 1994, The centrosome and its mode of inheritance: the reduction of the centrosome during gametogenesis and its restoration during fertilization. Dev Biol. 165:299–335.PubMedCrossRefGoogle Scholar
  44. Schatten, G., Hewitson, L., Simerly, C., Sutovsky, P., and Huszar, G., 1998, Cell and molecular biological challenges of ICSI: A.R.T. before science?. J Law Med Ethics. 26:29–37.PubMedCrossRefGoogle Scholar
  45. Schlegel, P.N., Palermo, G.D., Alikani, M, Adler, A., Reing, A.M., Cohen, J., and Rosenwaks Z., 1995, Micropuncture retrieval of epididymal sperm with in vitro fertilization: importance of in vitro micromanipulation techniques. Urology. 46:238–241.PubMedCrossRefGoogle Scholar
  46. Silber, S.J., Van Steirteghem, A.C., Liu, J., Nagy, Z., Tournaye, H., and Devroey, P., 1995, High fertilization and pregnancy rate after intracytoplasmic sperm injection with spermatozoa obtained from testicle biopsy. Hum Reprod. 10:148–152.PubMedCrossRefGoogle Scholar
  47. Silber, S.J., Johnson, L., Verheyen, G., and Van Steriteghem, A., 2000, Round spermatid injection. Fert Steril. 73:897–900.CrossRefGoogle Scholar
  48. Simerly, C., Wu, G., Zoran, S., Ord, T., Rawlins, R., Jones, J., Navara, C., Gerrity, M., Rinehart, J., Binor, Z., and Schatten, G., 1995, The paternal inheritance of the centrosome, the cell’s microtubuleorganizing center, in humans and the implications for infertility. Nature Med. 1:47–52.PubMedCrossRefGoogle Scholar
  49. Sofikitis, N., Mantzavinos, T., Loutradis, D., Yamamoto, Y., Tarlatzis, V., and Miyagawa, I., 1998, Ooplasmic injections of secondary spermatocytes for non-obstructive azoospermia. Lancet. 351:1177–1178.PubMedCrossRefGoogle Scholar
  50. St. John, J.C., Cooke, I.D., and Barratt, C.L., 1997, Mitochondrial mutations and male infertility. Nature Med. 3:124–125.PubMedGoogle Scholar
  51. Sutovsky, P., Hewitson, L., Simerly, C., Tengowski, M.W., Navara, C.S., Haavisto, A.J., and Schatten, G., 1996, Intracytoplasmic sperm injection for rhesus monkey fertilization results in unusual chromatin, cytoskeletal and membrane events, but eventually leads to pronuclear development and sperm aster assembly. Hum Reprod. 11:1703–1712.PubMedCrossRefGoogle Scholar
  52. Sutovsky, P., Oko, R., Hewitson, L., and Schatten, G., 1997, Binding of oocyte microvilli to the perinuclear theca of fertilizing sperm and subsequent theca removal constitute a previously unrecognized step in mammalian fertilization. Dev Biol. 188:75–84.PubMedCrossRefGoogle Scholar
  53. Tesarik, J., Mendoza, C., and Testart, J., 1995, Viable embryos from injection of round spermatids into oocytes. N Engl J Med. 333:525.PubMedCrossRefGoogle Scholar
  54. Terada, Y., Luetjens, C.M., Sutovsky, P., and Schatten, G., 2000, Atypical decondensation of the sperm nucleus, delayed replication of the male genome, and sex chromosome positioning following intracytoplasmic human sperm injection (ICSI) into golden hamster eggs: does ICSI itself introduce chromosomal anomalies?. Fertil Steril. 74:454–460.PubMedCrossRefGoogle Scholar
  55. te Velde, E.R., van Baar, A.L., and van Kooij, R.J., 1998, Concerns about assisted reproduction. Lancet. 351:1524–1525.CrossRefGoogle Scholar
  56. Tucker, M., Wright, G., Morton, C., Mayer, M.P., Ingargiola, P.E., and Jones, A.E., 1995, Practical evolution and application of direct intracytoplasmic sperm injection for male factor and idiopathic fertilization failure infertilities. Fertil Steril. 63:820–827.PubMedGoogle Scholar
  57. Van Steirteghem, A.C., Nagy, Z., Joris, H., Liu, J., Staessen, C., Smitz, J., Wisanto, A., and Devroey, P., 1993, High fertilization and implantation rates after intracytoplasmic sperm injection. Hum Reprod. 8: 1061–1066.PubMedGoogle Scholar
  58. Vogt, P.H., 1998, Human chromosome deletions in Yql 1, AZF candidate genes and male infertility: history and update. Mol Hum Reprod. 4:739–744.PubMedCrossRefGoogle Scholar
  59. Vogt, P.H., 1999, Risk of neurodegencrative diseases in children conceived by intracytoplasmic sperm injection?. Lancet. 354:611–612.PubMedCrossRefGoogle Scholar
  60. Wennerholm, U.B, Bergh, C, Hamberger, L., Lundin, K., Nilsson, L., Wikland, M., and Kallen, B., 2000, Incidence of congenital malformations in children born after ICSI. Hum Reprod. 15:944–948.PubMedCrossRefGoogle Scholar
  61. Wu, G., Simerly, C., Zoran, S., Funte, L.R., and Schatten, G., 1996, Microtubule and chromatin configurations during fertilization and early development in rhesus monkeys, and regulation by intracellular calcium ions. Biol Reprod. 55:260–270.PubMedCrossRefGoogle Scholar
  62. Yanagimachi, R., 1995, Is an animal model needed for intracytoplasmic sperm injection (ICSI) and other assisted reproductive technologies?. Hum Reprod. 10:2525–2526.PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2003

Authors and Affiliations

  • Laura Hewitson
    • 1
  • Calvin R. Simerly
    • 1
  • Gerald Schatten
    • 1
  1. 1.Pittsburgh Development CenterMagee-Womens Research Institute University of PittsburghPittsburghUSA

Personalised recommendations