J Vet Sci.
2011 Sep;12(3):267-272. 10.4142/jvs.2011.12.3.267.
Transmission electron microscopy for characterization of acrosomal damage after Percoll gradient centrifugation of cryopreserved bovine spermatozoa
- Affiliations
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- 1Department of Preventive Veterinary Medicine and Animal Reproduction, School of Agrarian Sciences and Veterinary Medicine, Universidade Estadual Paulista, Jaboticabal 14884-900, Brazil. leticiazoccolaro@yahoo.com.br
- 2Laboratory of Histology, Department of Morphology, Biomedical Sciences Institute, Universidade Federal de Uberlandia, Uberlandia 38400-902, Brazil.
- 3Department of Animal Reproduction, School of Veterinary Medicine, Universidade Federal de Uberlandia, Uberlandia 38400-902, Brazil.
- 4Laboratory of Semen Biotechnology and Andrology, Department of Animal Reproduction, Veterinary Medicine and Animal Science School, Universidade de Sao Paulo, Pirassununga 13630-000, Brazil.
- KMID: 1067399
- DOI: http://doi.org/10.4142/jvs.2011.12.3.267
Abstract
- The objective of this study was to characterize acrosomal ultrastructure following discontinuous Percoll gradient centrifugation of cryopreserved bovine sperm. Semen was collected from six bulls of different breeds and three ejaculates per bull were evaluated. Frozen semen samples were thawed and the acrosomal region of sperm cells was evaluated by transmission electron microscopy (TEM) before (n = 18) and after (n = 18) Percoll centrifugation. The evaluation of 20 sperm heads from each of the 36 samples analyzed ensured that a large number of cells were investigated. The data were subjected to analysis of variance at a level of significance of 5%. Percoll centrifugation reduced the percentage of sperm exhibiting normal acrosomes (from 61.77 to 30.24%), reduced the percentage of sperm presenting atypical acrosome reactions (from 28.38 to 4.84%) and increased the percentage of sperm exhibiting damage in the acrosome (from 6.14 to 64.26%). The percentage of sperm with typical acrosome reactions was not significantly different before (3.70%) and after (0.67%) centrifugation. TEM distinguished four different types of acrosomal status and enabled ultrastructural characterization of acrosomal injuries. The percentage of sperm exhibiting normal acrosomes decreased and damage in the acrosome was the most frequent acrosomal injury with the Percoll gradient centrifugation protocol utilized.
Keyword
MeSH Terms
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Acrosome/*pathology/ultrastructure
Animals
Cattle/*physiology
Cell Membrane/*pathology/ultrastructure
Cell Separation/veterinary
Centrifugation, Density Gradient/veterinary
Cryopreservation/veterinary
Male
Microscopy, Electron, Transmission/veterinary
Povidone/*adverse effects
Silicon Dioxide/*adverse effects
Spermatozoa/pathology/ultrastructure
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