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Research article - Peer-reviewed, 2000

Evaluation of chilled and frozen-thawed canine spermatozoa using a zona pellucida binding assay

Ström Holst, Bodil; Larsson, Birgitta; Linde-Forsberg, Catharina; Rodriguez-Martinez, Heriberto

Abstract

Zona pellucida binding assays provide information about the fertilizing ability of spermatozoa. A zona-binding assay for canine spermatozoa using intact, denuded homologous oocytes has not been evaluated previously, in the present study, an assay using canine oocytes derived from frozen-thawed ovaries was evaluated using three types of semen: fresh untreated; killed; and a 50:50 mixture of untreated and killed spermatozoa. The assays were performed on 3 x 20 oocytes for each sperm treatment, using semen from pooled ejaculates (0.5 x 10(6) spermatozoa in each 50 Cll droplet containing five oocytes). There was a significant difference (P < 0.001) between all treatments. Thereafter, the same procedure was used to evaluate methods of chilling and freeze-thawing of canine semen. There was a trend (P = 0.067) for more sperm binding after 1 day of chilling compared with after 4 days of chilling. Semen samples frozen using an extender (with or without the addition of Equex STM paste) were evaluated. Equex had a significant (P = 0.034) positive effect on the capacity of the spermatozoa to bind to the zona pellucida. In conclusion, the addition of a zona pellucida binding assay to established in vitro tests should give a better estimate of the damage caused by the various procedures when developing new techniques for chilling and freeze-thawing. Furthermore, the present study showed that chilling for 4 days tended to reduce the zona-binding capacity of the spermatozoon, and that Equex STM paste had a beneficial effect on the capacity of the frozen-thawed spermatozoon to bind to the zona pellucida.

Published in

Journal Of Reproduction And Fertility
2000, Volume: 119, number: 2, pages: 201-206
Publisher: JOURNALS OF REPRODUCTION FERTILITY LTD

      SLU Authors

    • Ström Holst, Bodil

      • Department of Obstetrics and Gynaecology, Swedish University of Agricultural Sciences
      • Larsson, Birgitta

        • Department of Obstetrics and Gynaecology, Swedish University of Agricultural Sciences
        • Linde-Forsberg, Catharina

          • Department of Obstetrics and Gynaecology, Swedish University of Agricultural Sciences
          • Rodriguez, Heriberto

            • Department of Obstetrics and Gynaecology, Swedish University of Agricultural Sciences

          UKÄ Subject classification

          Clinical Science

          Publication identifier

          DOI: https://doi.org/10.1530/reprod/119.2.201

          Permanent link to this page (URI)

          https://res.slu.se/id/publ/43145