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  • Cryopreservation of Boar Semen
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III: Ultrastructure of Boar Spermatozoa Frozen Ultra-Rapidly at Various Stages of Conventional Freezing and Thawing

Djupfrysning av galtspermier. III: Ultrastrukturen hos galtspermier, som frysts med ultrahög hastighet vid olika stadier av den konventionella infrysning- upptinings proceduren

Acta Veterinaria Scandinavica volume 32, pages 463–471 (1991)Cite this article

Abstract

Ejaculated boar spermatozoa subjected to a conventional freezing and thawing process, were ultra-rapidly fixed, freeze-substituted and examined by electron microscopy to monitor the presence of real or potential intracellular ice and the degree of cell protection attained with the different extenders used during the process. Numerous ice crystal marks representing the degree of hydration of the cells were located in the perinuclear space of those spermatozoa not in proper contact with the extender containing glycerol (i.e. prior to freezing). The spermatozoa which were in proper contact with the extenders presented a high degree of preservation of the acrosomes, plasma membranes as well as the nuclear envelopes. No ice marks were detected in acrosomes before thawing, indicating that the conventional assayed cryopreservation method provided a good protection against cryoinjury. The presence of acrosomal changes (internal vesiculization, hydration and swelling) in thawed samples however, raises serious questions about the thawing procedure employed.

Sammanfattning

Elakulerade galtspermier, som utsatts för konventionell infrysning och upptining, fixerades med ultrarapid hastighet, fryssubstituerades och under-söktes elektronmikroskopiskt för bestämning av närvaron av verklig eller potentiell intracellulär is och graden av cellskada uppkommen under processen. Tallrika spår av iskristaller, represente-rande graden av hydrering i cellerna, var lokaliserade til perinukleära utrymmet hos de spermier som inte var i tillräcklig kontakt med den glyceroltillsatta spädningsvätskan. De spermier, som var i tillräcklig kontakt med den glyceroltillsatta spädningsvätskan, visade i hög grad intakta akrosomer, plasmamembraner och kärnmembraner. Inga spår av iskristaller upptäcktes i akrosomerna före upptining, vilket visar att den konventionella djupfrysningsmetoden gav ett gott skydd mot frysskador. Förekomsten av akrosomskador (inre vesikulering, hydrering, avsvällning) i upptinade prover, reser allvarliga frågor om den använda tiningsproceduren.

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Authors and Affiliations

  1. Department of Anatomy and Histology and Department of Obstetrics and Gynaecology, Faculty of Veterinary Medicine, Swedish University of Agricultural Sciences, P. O. Box 7039, S-75007, Uppsala, Sweden

    C. O. Bwanga, H. Ekwall & H. Rodriguez-Martinez

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  1. C. O. Bwanga
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  2. H. Ekwall
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  3. H. Rodriguez-Martinez
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Bwanga, C.O., Ekwall, H. & Rodriguez-Martinez, H. III: Ultrastructure of Boar Spermatozoa Frozen Ultra-Rapidly at Various Stages of Conventional Freezing and Thawing. Acta Vet Scand 32, 463–471 (1991). https://doi.org/10.1186/BF03546946

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Acta Veterinaria Scandinavica

ISSN: 1751-0147