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This Article Full Text Full Text (PDF) All Versions of this Article: 71/1/28    most recent biolreprod.103.024281v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal My Folders Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Martin, G. Articles by Levy, R. Search for Related Content PubMed PubMed Citation Articles by Martin, G. Articles by Levy, R. Agricola Articles by Martin, G. Articles by Levy, R.

Cryopreservation Induces an Apoptosis-Like Mechanism in Bull Sperm

Laboratoire de Biologie de la Reproduction—GIMAP,² Hôpital Nord, 42055 Saint-Etienne, France INSERM U418—INRA UMR1245,³ Hôpital Debrousse, 69322 Lyon, France Centre Commun de Cytométrie en Flux,⁴ Université Jean Monnet, 42023 Saint-Etienne Cedex 2, France

Cryopreservation induces many changes in sperm cells, including membrane disorders and cell death. We tested the hypothesis that apoptosis, a form of programmed cell death, can contribute to the fatal effect of cryopreservation on sperm cells. A multiparametric study of apoptosis on bovine sperm is proposed, using flow cytometry, including mitochondrial membrane potential (_m), caspase activation, membrane permeability, nucleus condensation, DNA fragmentation, and phosphatidylserine (PS) externalization. The relevance of each test was first validated on a human somatic cell line, U937. Cryopreservation and/or thawing induced significant changes in all apoptotic markers in living bull sperm cells except those concerning the nucleus. After cryopreservation, 44.9% ± 17% (vs. 11.3% ± 10.6% before cryopreservation) of sperm cells showed low _m, 12% ± 6.3% (vs. 2.2% ± 1.0% before) contained active caspases, and 10.8% ± 5.8% (vs. 1.4% ± 1.1% before) exhibited high membrane permeability. However, cryopreservation had no effect on DNA fragmentation (9.1% ± 7.7% before vs. 11.1% ± 5.7% after cryopreservation) or on nucleus condensation (46% ± 12.7% before vs. 43.8% ± 13.1% after). Cryopreservation acts as an apoptotic mechanism inducer in bovine sperm cells, where the earliest but not the latest features of cells undergoing apoptosis occur. We have named this abortive process an apoptosis-like phenomenon.

¹ Correspondence. FAX: 33 0 4 77 82 84 61; rachel.levy{at}chu-st-etienne.fr

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