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Chapter 10 - Capacitation and Acrosome Reaction: Histochemical Techniques to Determine Acrosome Reaction

Published online by Cambridge University Press:  05 April 2021

Ashok Agarwal
The Cleveland Clinic Foundation, Cleveland, OH
Ralf Henkel
University of the Western Cape, South Africa
Ahmad Majzoub
Hamad Medical Corporation, Doha
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Fertilization is an exceptionally specific cell recognition event that represents the culmination of a complex sequence of morphological and functional maturational events. In the case of the male gamete, this process is initiated by the commitment of spermatogonial stem cells to differentiate, sequentially forming spermatogonia, spermatocytes and eventually spermatozoa that are released into the lumen of the seminiferous tubules [1]. In addition to meiotic divisions, this process encompasses extensive cytoplasmic, organelle and nuclear remodeling events, thus establishing the unique and highly polarized architecture of the mature spermatozoon. A key aspect of this phase of development is the modification and repositioning of the Golgi apparatus to form a highly specialized secretory organelle, known as the acrosome, overlying the anterior aspect of the sperm head. Upon release from the testes the functionally immature spermatozoa enter the epididymis where they are progressively remodeled and acquire both motility and the potential to fertilize an oocyte [2]. This potential is eventually realized after passage through the female reproductive tract whereupon the ejaculated cells complete a suite of biochemical and biophysical changes known as capacitation [3]. These successive phases of functional maturation culminate in the acquired ability to release the acrosomal contents, during an event known as the acrosome reaction. This unique exocytotic event facilitates sperm passage through the outer vestments of the oocyte and is essential for successful in vivo fertilization in all mammalian species, including the human [4]. Consequently, failure of acrosomal exocytosis represents a common etiology in defective spermatozoa of male infertility patients that have failed in vitro fertilization (IVF) in a clinical setting; accounting for as much as 29 percent of unexplained male infertility cases [5, 6]. Much of our current mechanistic understanding of the acrosome reaction is grounded in the ability to stimulate this process in vitro using simple chemically defined media and the application of pharmacological interventions, and/or transgenic mouse models. Here, we discuss the biological significance of the acrosome reaction and the application of histochemical techniques that have been developed to study the progression and completion of this critical physiological event.

Publisher: Cambridge University Press
Print publication year: 2021

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Hermo, L, Pelletier, RM, Cyr, DG, Smith, CE. Surfing the wave, cycle, life history, and genes/proteins expressed by testicular germ cells. Part 2: changes in spermatid organelles associated with development of spermatozoa. Microsc Res Tech 2010; 73: 279319.CrossRefGoogle ScholarPubMed
Aitken, RJ, Nixon, B, Lin, M, Koppers, AJ, Lee, YH, Baker, MA. Proteomic changes in mammalian spermatozoa during epididymal maturation. Asian J Androl 2007; 9: 554–64.CrossRefGoogle ScholarPubMed
Nixon, B, Bromfield, EG. (2018) Sperm capacitation. In Skinner, M.K., ed., The Encyclopedia of Reproduction. Cambridge, MA: Academic Press, Elsevier, pp. 272–8.Google Scholar
Baker, HW, Liu, DY, Garrett, C, Martic, M. The human acrosome reaction. Asian J Androl 2000; 2: 172–8.Google ScholarPubMed
Liu, DY, Clarke, GN, Martic, M, Garrett, C, Baker, HW. Frequency of disordered zona pellucida (ZP)-induced acrosome reaction in infertile men with normal semen analysis and normal spermatozoa-ZP binding. Hum Reprod 2001; 16: 1185–90.Google Scholar
Liu, DY, Baker, HW. Disordered zona pellucida-induced acrosome reaction and failure of in vitro fertilization in patients with unexplained infertility. Fertil Steril 2003; 79: 7480.CrossRefGoogle ScholarPubMed
Guyonnet, B, Egge, N, Cornwall, GA. Functional amyloids in the mouse sperm acrosome. Mol Cell Biol 2014; 34: 2624–34.CrossRefGoogle ScholarPubMed
Guyonnet, B, Zabet-Moghaddam, M, SanFrancisco, S, Cornwall, GA. Isolation and proteomic characterization of the mouse sperm acrosomal matrix. Mol Cell Proteomics 2012; 11: 758–74.Google Scholar
Zhou, W, Anderson, AL, Turner, AP, De Iuliis, GN, McCluskey, A, McLaughlin, EA, Nixon, B. Characterization of a novel role for the dynamin mechanoenzymes in the regulation of human sperm acrosomal exocytosis. Mol Hum Reprod 2017; 23: 657–73.CrossRefGoogle ScholarPubMed
Buffone, MG, Foster, JA, Gerton, GL. The role of the acrosomal matrix in fertilization. Int J Dev Biol 2008; 52: 511–22.Google Scholar
Bourne, H, Liu, DY, Clarke, GN, Baker, HW. Normal fertilization and embryo development by intracytoplasmic sperm injection of round-headed acrosomeless sperm. Fertil Steril 1995; 63: 1329–32.CrossRefGoogle ScholarPubMed
Nagy, ZP, Liu, J, Joris, H, Verheyen, G, Tournaye, H, Camus, M, Derde, MC, Devroey, P, Van Steirteghem, AC. The result of intracytoplasmic sperm injection is not related to any of the three basic sperm parameters. Hum Reprod 1995; 10: 1123–9.CrossRefGoogle ScholarPubMed
Stival, C, Puga Molina Ldel, C, Paudel, B, Buffone, MG, Visconti, PE, Krapf, D. Sperm capacitation and acrosome reaction in mammalian sperm. Adv Anat Embryol Cell Biol 2016; 220: 93106.Google Scholar
Nixon, B, MacIntyre, DA, Mitchell, LA, Gibbs, GM, O'Bryan, M, Aitken, RJ. The identification of mouse sperm-surface-associated proteins and characterization of their ability to act as decapacitation factors. Biol Reprod 2006; 74: 275–87.Google Scholar
Buffone, MG, Hirohashi, N, Gerton, GL. Unresolved questions concerning mammalian sperm acrosomal exocytosis. Biol Reprod 2014; 90: 112.CrossRefGoogle ScholarPubMed
La Spina, FA, Puga Molina, LC, Romarowski, A, Vitale, AM, Falzone, TL, Krapf, D, Hirohashi, N, Buffone, MG. Mouse sperm begin to undergo acrosomal exocytosis in the upper isthmus of the oviduct. Dev Biol 2016; 411: 172–82.Google Scholar
Hirohashi, N, Gerton, GL, Buffone, MG. Video imaging of the sperm acrosome reaction during in vitro fertilization. Commun Integr Biol 2011; 4: 471–6.Google Scholar
Liu, DY, Garrett, C, Baker, HW. Acrosome-reacted human sperm in insemination medium do not bind to the zona pellucida of human oocytes. Int J Androl 2006; 29: 475–81.Google Scholar
Buffone, MG, Rodriguez-Miranda, E, Storey, BT, Gerton, GL. Acrosomal exocytosis of mouse sperm progresses in a consistent direction in response to zona pellucida. J Cell Physiol 2009; 220: 611–20.Google Scholar
Liu, DY, Baker, HW. Evaluation and assessment of semen for IVF/ICSI. Asian J Androl 2002; 4: 281–5.Google Scholar
Liu, DY, Stewart, T, Baker, HW. Normal range and variation of the zona pellucida-induced acrosome reaction in fertile men. Fertil Steril 2003; 80: 384–9.CrossRefGoogle ScholarPubMed
Talbot, P, Chacon, RS. A triple-stain technique for evaluating normal acrosome reactions of human sperm. J Exp Zool 1981; 215: 201–8.Google Scholar
Mitchell, LA, Nixon, B, Aitken, RJ. Analysis of chaperone proteins associated with human spermatozoa during capacitation. Mol Hum Reprod 2007; 13: 605–13.CrossRefGoogle ScholarPubMed
Aitken, RJ, Buckingham, DW, Fang, HG. Analysis of the responses of human spermatozoa to A23187 employing a novel technique for assessing the acrosome reaction. J Androl 1993; 14: 132–41.Google Scholar
Didion, BA, Dobrinsky, JR, Giles, JR, Graves, CN. Staining procedure to detect viability and the true acrosome reaction in spermatozoa of various species. Gamete Res 1989; 22: 51–7.CrossRefGoogle ScholarPubMed
Larson, JL, Miller, DJ. Simple histochemical stain for acrosomes on sperm from several species. Mol Reprod Dev 1999; 52: 445–9.Google Scholar
Esterhuizen, AD, Franken, DR, Lourens, JG, van Rooyen, LH. Clinical importance of zona pellucida-induced acrosome reaction and its predictive value for IVF. Hum Reprod 2001; 16: 138–44.Google Scholar
Aitken, RJ, Thatcher, S, Glasier, AF, Clarkson, JS, Wu, FC, Baird, DT. Relative ability of modified versions of the hamster oocyte penetration test, incorporating hyperosmotic medium or the ionophore A23187, to predict IVF outcome. Hum Reprod 1987; 2: 227–31.CrossRefGoogle ScholarPubMed
Liu, DY, Baker, HW. A simple method for assessment of the human acrosome reaction of spermatozoa bound to the zona pellucida: lack of relationship with ionophore A23187-induced acrosome reaction. Hum Reprod 1996; 11: 551–7.Google Scholar
Risopatron, J, Pena, P, Miska, W, Sanchez, R. Evaluation of the acrosome reaction in human spermatozoa: comparison of cytochemical and fluorescence techniques. Andrologia 2001; 33: 63–7.Google Scholar
Aitken, RJ. Sperm function tests and fertility. Int J Androl 2006; 29: 6975.Google Scholar

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