Skip to main content
SpringerLink
Log in

Acid extraction of acrosin from human spermatozoa pretreated by different physicochemical methods

  • Original Contributions
  • Published:
Archives of Dermatological Research Aims and scope Submit manuscript

Summary

Washed human spermatozoa were subjected to different physicochemical methods, followed by acid extraction of the sperm acrosomes to dissolve acrosin. The acrosin activity of the sperm pellet and the supernatant was determined by benzoyl arginine ethyl ester/alcohol dehydrogenase (BAEE/ADH) assay to calculate the total acrosin activity in m U/106 spermatozoa. Membrane-active and zymogen-activating agents increased the total acrosin activity 50%–200% compared to acid extraction alone. Similar results were obtained by osmotic shock, sonication and treatment with glass beads. Snap freezing of unprotected spermatozoa in liquid nitrogen yielded a fivefold increase in total acrosin activity, thus demonstrating that this is the method of choice for optimal acrosin extraction. The possibility is discussed as to whether acrosomal membrane alterations with improved solubilization of membranebound acrosin and/or conformational changes and/or zymogen activation are responsible for the considerable increase observed in acrosin activity.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Bernstein MH, Teichman RJ (1973) A chemical procedure for extraction of the acrosomes of mammalian spermatozoa. J Reprod Fertil 33:239–244

    PubMed  Google Scholar 

  2. Berruti G (1980) Multiple forms of boar, bull and human acrosin. Arch Androl 5:267–277

    PubMed  Google Scholar 

  3. Bhattacharyya AK, Zaneveld LJD (1978) Release of acrosin and acrosin inhibitor from human spermatozoa. Fertil Steril 30:70–78

    PubMed  Google Scholar 

  4. Brown CR, Hartree EF (1976) Effects of acrosin inhibitors on the soluble and membrane-bound forms of ram acrosin, and a reappraisal of the role of the enzyme in fertilzation. Hoppe Seylers Z Physiol Chem 357:57–65

    PubMed  Google Scholar 

  5. Churg A, Zaneveld LJD, Schumacher GFB (1974) Detergent treatment of human and rabbit spermatozoa: ultrastructural changes and release of midpiece enzymes. Biol Reprod 10:429–437

    PubMed  Google Scholar 

  6. Clermont Y, Glegg RE, Leblond CP (1955) Presence of carbohydrates in the acrosomes of the guinea pig spermatozoon. Exp Cell Res 8:452–457

    Google Scholar 

  7. Dudkiewicz AB, Srivastava PN, Yang CH, Williams WL (1979) Extraction of human and rabbit acrosomes: a comparison of sequential and sonication methods. Andrologia 11:355–366

    PubMed  Google Scholar 

  8. Filip C, Fletcher G, Wulff J, Earhart CF (1973) Solubilization of the cytoplasmic membrane of Escherichia coli by ionic detergent sodium-laurly sarcosinate. J Bacteriol 115:717–722

    PubMed  Google Scholar 

  9. Fléchon J-E, Huneau D, Brown CR, Harrison RAP (1977) Immunocytochemical localization of acrosin in the anterior segment of the acrosomes of ram, boar and bull spermatozoa. Ann Biol Anim Biochim Biophys 17:749–758

    Google Scholar 

  10. Fritz H, Förg-Brey B, Fink E, Schiessler H, Schirren C (1972) Humanakrosin Gewinnung und Eigenschaften. Hoppe Seylers Z Physiol Chem 353:1943–1949

    PubMed  Google Scholar 

  11. Fritz H, Schiessler H, Schill W-B, Tschesche H, Heimburger N, Waller O (1975a) Low molecular weight proteinase (acrosin) inhibitors from human and boar seminal plasma and spermatozoa and human cervical mucus-isolation, properties and biological aspects. in: Reich E, Rifkin DB, Shaw E (eds) Proteases and biological control. Cold Spring Harbor Laboratory, Cold Spring Harbor, USA, pp 737–766

    Google Scholar 

  12. Fritz H, Schleuning W-D, Schiessler H, Schill W-B, Wendt V, Winkler G (1975b) Boar, bull and human sperm acrosin-isolation, properties and biological aspects. In: Reich E, Rifkin DB, Shaw E (eds) Proteases and biological control Cold Spring Harbor Laboratory, Cold Spring Harbor USA, pp 715–735

    Google Scholar 

  13. Gaddum P, Blandau RJ (1970) Proteolytic reaction of mammalian spermatozoa on gelatin membranes. Science 170:749–751

    PubMed  Google Scholar 

  14. Garbara B, Gledhill BL, Croce CM, Cesarini JP, Koprowski H (1973) Ultrastructure of rabbit spermatozoa after treatment with lysolecithin and in the presence of hamster somatic cells. Proc Soc Exp Biol Med 143:1120–1124

    PubMed  Google Scholar 

  15. Gilboa E, Elkana Y Rigbi M (1973) Purification and properties of human acrosin. Eur J Biochem 39:85–92

    PubMed  Google Scholar 

  16. González-Angulo A, Martinez-Zedillo G, Delgado N Martínez-Manautou J (1971) Effect of hypertonic calcium chloride upon the fine structure of human spermatozoa. Int J Fertil 16:161–168

    PubMed  Google Scholar 

  17. Goodpasture JC, Polakoski KL, Zaneveld LJD (1980) Acrosin, proacrosin, and acrosin inhibitor of human spermatozoa: extraction, quantitation, and stability, J Androl 1:16–27

    Google Scholar 

  18. Harrison RAP, Brown CR (1979) The zymogen form of acrosin in testicular, epididymal, and ejaculated spermatozoa from ram. Gamete Res 2:75–87

    Google Scholar 

  19. Hartree EF, Srivastava PN (1965) Chemical composition of the acrosomes of ram spermatozoa. J Reprod Fertil 9:47–60

    Google Scholar 

  20. Hathaway RR, Hartree EF (1963) Observations on the mammalian acrosome: experimental removal of acrosomes from ram and bull spermatozoa. J Reprod Fertil 5:225–232

    PubMed  Google Scholar 

  21. Hernández-Montes H, Iglesias G, Mújica A (1973) Selective solubilization of mammalian spermatozoa structures. Exp Cell Res 76:437–440

    PubMed  Google Scholar 

  22. Johnson LA, Pursel VG, Chaney N, Garner D (1976) Comparison of several extraction procedures for boar spermatozoal acrosin. Biol Reprod 15:79–83

    PubMed  Google Scholar 

  23. McRorie RA, Williams WL (1974) Biochemistry of mammalian fertilization. Annu Rev Biochem 43:777–803

    Article  PubMed  Google Scholar 

  24. Morton DB (1975) Acrosmal enzymes: immunochemical localization of acrosin and hyaluronidase in ram spermatozoa. J Reprod Fertil 45:375–378

    PubMed  Google Scholar 

  25. Müller-Esterl W, Fritz H (1980) Interaction of boar acrosin with detergents. Hoppe Seylers Z Physiol Chem 361:1673–1682

    PubMed  Google Scholar 

  26. Mukerji SK, Meizel S (1975) Conversion of rabbit testis proacrosin to acrosin. FEBS Lett 54:269–273

    Article  PubMed  Google Scholar 

  27. Multamäki S (1973) Isolation of pure acrosomes by subcellular fractionation of bull spermatozoa. Int J Fertil 18:193–205

    PubMed  Google Scholar 

  28. Multamäki S, Niemi N (1972) Trypsin-like proteolytic activity in an acrosomal extract of bull spermatozoa. Int J Fertil 17:43–52

    PubMed  Google Scholar 

  29. Multamäki S, Suominen J (1976) Distribution and removal of the acrosin of bull spermatozoa. Int J Fertil 21:69–81

    Google Scholar 

  30. Parrish RF, Polakoski KL (1977) Effect of polyamines on the activity of acrosin and the activation of proacrosin. Biol Reprod 17:417–422

    PubMed  Google Scholar 

  31. Pedersen H (1972) The acrosome of the human spermatozoon: a new method for its extraction, and an analysis of its trypsin-like enzyme activity. J Reprod Fertil 31:99–107

    PubMed  Google Scholar 

  32. Polakoski KL, Zaneveld LJD, Williams WL (1972) Purification of a proteolytic enzyme from rabbit acrosomes. Biol Reprod 6:23–39

    PubMed  Google Scholar 

  33. Polakoski KL, Zahler WL, Paulson JD (1977) Demonstration of proacrosin and quantitation of acrosin in ejaculated human spermatozoa. Fertil Steril 28:668–670

    PubMed  Google Scholar 

  34. Reynolds JA, Trayer H (1971) Solubility of membrane proteins in aqueous media. J Biol Chem 246:7337–7342

    PubMed  Google Scholar 

  35. Schill W-B (1973) Acrosin activity in human spermatozoa: methodological investigations. Arch Dermol Forsch 248:257–273

    Google Scholar 

  36. Schill W-B (1974a) Quantitation determination of acrosin activity in human spermatozoa. Fertil Steril 25:703–712

    PubMed  Google Scholar 

  37. Schill W-B (1974b) The influence of glycerol on the extractability of acrosin from human spermatozoa. Hoppe Seylers Z Physiol Chem 355:225–228

    PubMed  Google Scholar 

  38. Schill W-B (1975) Acrosin activity of cryo-preserved human spermatozoa. Fertil Steril 26:711–720

    PubMed  Google Scholar 

  39. Schill W-B (1976) Verhalten der Akrosinaktivität von Kryosperma. In: Kaden R, Lübke F, Schirren C (eds) Fortschritte der Fertilitätsformung. III. Grosse, Berlin, pp 259–265

    Google Scholar 

  40. Schill W-B (1981) Acrosin and seminal plasma proteinase inhibitors in the diagnostic workup of male infertility. In: Insler V, Bettendorf G (eds) Advances in diagnosis and treatment of infertility. Elsevier/North-Holland New York, pp 321–337

    Google Scholar 

  41. Schill W-B, Fritz H (1976) Enhancement of sperm acrosin activity by glycerol-pretreatment-quantitative estimations. Arch Dermatol Res 256:1–11

    PubMed  Google Scholar 

  42. Schill W-B, Wolff HH (1981) Ultrastructure of human spermatozoa in the presence of the spermicide nonoxinol-9 and a vaginal contraceptive containing nonoxinol-9. Andrologia 13:42–49

    PubMed  Google Scholar 

  43. Schill W-B, Schleuning WD, Fritz H, Wendt V, Heimburger N (1975) Immunofluorescent localization of acrosin in spermatozoa by boar acrosin antibodies. Naturwissenschaften 62:540–541

    PubMed  Google Scholar 

  44. Schill W-B, Feifel M, Fritz H, Hammerstein J (1981) Inhibitors of acrosomal proteinase as antifertility agents. A problem of acrosomal membrane permeability. Int J Androl 4:25–38

    PubMed  Google Scholar 

  45. Schirren C (1971) Praktische Andrologie. Hartmann, Berlin

    Google Scholar 

  46. Schleuning W-D, Hell R, Fritz H (1976) Multiple forms of human acrosin. Isolation and properties. Hoppe Seylers Z Physiol Chem 357:855–865

    PubMed  Google Scholar 

  47. Srivastava PN (1973) Removal of acrosomes of ram and rabbit spermatozoa. J Reprod Fertil 33:323–326

    PubMed  Google Scholar 

  48. Srivastava PN, Munnell JF, Yang CH, Foley CW (1974) Sequential release of acrosomal membranes and acrosomal enzymes of ram spermatozoa. J Reprod Fertil 36: 363–372

    PubMed  Google Scholar 

  49. Stambaugh R, Buckley J (1968) Zona pellucida dissolution enzymes of the rabbit sperm head. Science 161:585–586

    PubMed  Google Scholar 

  50. Stambaugh R, Buckley J (1969) Identification and subcellular localization of the enzymes affecting penetration of the zona pellucide by rabbit spermatozoa. J Reprod Fertil 19:423–432

    PubMed  Google Scholar 

  51. Stambaugh R, Buckley J (1970) Comparative studies of the acrosomal enzymes of rabbit, rhesus monkey, and human spermatozoa. Biol Reprod 3:275–282

    PubMed  Google Scholar 

  52. Stambaugh R, Smith M (1973) A comparison of several extraction procedures for rabbit acrosomal enzymes. J Reprod Fertil 35:127–130

    PubMed  Google Scholar 

  53. Teichman RJ, Bernstein MH (1969) A morphological and biochemical comparison on rabbit, human, and bull acrosomes. J Cell Biol 43:144a-145a

    Google Scholar 

  54. Tobias PS, Schumacher GFB (1976) The extraction of acrosin from human spermatozoa. Biol Reprod 15:187–194

    PubMed  Google Scholar 

  55. Trautschold J, Werle E (1961) Spektrophotometrische Bestimmung des Kallikreins und seiner Inaktivatoren. Hoppe Seylers Z Physiol Chem 325:48–59

    PubMed  Google Scholar 

  56. Wolff HH, Schill W-B (1975) Ultrastructural study of human sperm acrosome during acetic acid extraction of acrosin. J Reprod Fertil 42:385–387

    PubMed  Google Scholar 

  57. Wooding FBP (1973) The effect of triton X-100 on the ultrastructure of ejaculated bovine sperm. J Ultrastruc Res 42:502–516

    Google Scholar 

  58. Wooding FBP (1975) Studies on the mechanism of the hyamine-induced acrosome reaction in ejaculated bovine spermatozoa. J Reprod Fertil 44:185–192

    PubMed  Google Scholar 

  59. Zaneveld LJD, Dragoje BM, Schumacher GFB (1972) Acrosomal proteinase and proteinase inhibitors of human spermatozoa. Science 177:702–703

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Dermatologische Klinik und Poliklinik der Ludwig-Maximilians-Universität München, Frauenlobstr. 9-11, D-8000, München 2, Federal Republic of Germany

    W.-B. Schill & M. Feifel

  2. Abteilung für Klinische Chemie und Klinische Biochemie an der Chirurgischen Klinik der Ludwig-Maximilians-Universität München, Nußbaumstr. 20, D-8000, München 2, Federal Republic of Germany

    H. Fritz

Authors
  1. W.-B. Schill
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. M. Feifel
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. H. Fritz
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Dedicated to Prof. Dr. Dr. h.c. Otto Braun-Falco in honour of his 60th birthday

Rights and permissions

Reprints and permissions

About this article

Cite this article

Schill, WB., Feifel, M. & Fritz, H. Acid extraction of acrosin from human spermatozoa pretreated by different physicochemical methods. Arch Dermatol Res 273, 273–285 (1982). https://doi.org/10.1007/BF00409256

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00409256

Key words

Search

Navigation