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Article << Previous     |     Next >>   Contents Vol 23(7)

Colloid centrifugation removes seminal plasma and cholesterol from boar spermatozoa

R. Kruse A D, P. C. Dutta B and J. M. Morrell C E

A Clinical Sciences, Swedish University of Agricultural Sciences (SLU), Box 7054, SE-75007 Uppsala, Sweden.
B Department of Food Science, Uppsala BioCenter, Swedish University of Agricultural Sciences, Box 7051, SE-75007 Uppsala, Sweden.
C Division of Reproduction, Clinical Sciences, Swedish University of Agricultural Sciences, Box 7054, SE-75007 Uppsala, Sweden.
D Present address: Department of Clinical Medicine, School of Health and Medical Sciences, Örebro University, Örebro, Sweden.
E Corresponding author. Email: jane.morrell@slu.se

Reproduction, Fertility and Development 23(7) 858-865 http://dx.doi.org/10.1071/RD10260
Submitted: 7 October 2011  Accepted: 28 March 2011   Published: 19 August 2011

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The objective of the present study was to investigate the effect of Single-Layer Centrifugation (SLC) on boar spermatozoa, namely the effect of removal of seminal plasma proteins and cholesterol from the surface of spermatozoa. The presence of porcine seminal plasma proteins I and II (PSP-I/PSP-II) before and after SLC was studied using immunofluorescence, whereas the removal of cholesterol was shown qualitatively by thin-layer chromatography (TLC). Finally, the integrity of the sperm plasma membrane was observed by electron microscopy. It was shown that the seminal plasma proteins PSP-I and -II were removed from spermatozoa during SLC but could be restored by adding seminal plasma to the SLC-selected sperm samples. Some cholesterol was also lost from the spermatozoa during SLC but the plasma membrane itself appeared to be morphologically intact. Further studies are underway to examine the relevance of these findings to boar sperm cryopreservation and sperm fertility.

Additional keywords: PSP-I, PSP-II, SLC.


Amorim, E. A., Graham, J. K., Spizziri, B., Meyers, M., and Torres, C. A. (2009). Effect of cholesterol or cholesteryl conjugates on the cryosurvival of bull sperm. Cryobiology 58, 210–214.
CrossRef | CAS | PubMed |

Arienti, G., Carlini, E., and Palmerinin, C. A. (1997). Fusion of human sperm to prostasomes at acidic pH. J. Membr. Biol. 155, 89–94.
CrossRef | CAS | PubMed |

Avery, B., and Greve, T. (1995). Impact of Percoll on bovine spermatozoa used for in vitro insemination. Theriogenology 44, 871–878.
| CAS | PubMed |

Bligh, E. G., and Dyer, W. J. (1959). A rapid method of total lipid extraction and purification. Can. J. Biochem. Physiol. 37, 911–917.
CrossRef | CAS | PubMed |

Blomqvist, G., Persson, M., Wallgren, M., Wallgren, P., and Morrell, J. M. (2011). Removal of virus from boar semen spiked with porcine circovirus type 2. Anim. Reprod. Sci. 126, 108–114.
| PubMed |

Caballero, I., Vazquez, J. M., Garcia, E. M., Roca, J., Martinez, E. A., Calvete, J. J., Sanz, L., Ekwall, H., and Rodriguez-Martinez, H. (2006). Immunolocalization and possible functional role of PSP-I/PSP-II heterodimer in highly extended boar spermatozoa. J. Androl. 27, 766–773.
CrossRef | CAS | PubMed |

Calvete, J. J., Sanz, L., Garcia, E. M., Caballero, I., Parilla, I., Martinez, E., Roca, J., Vazquez, J. M., Saravia, F., Wallgren, M., Johannisson, A., and Rodriguez-Martinez, H. (2005). On the biological function of boar spermadhesin PSP-I/PSP-II. Reprod. Domest. Anim. 40, 331..

Causnicu, P. S., Cohen, D. J., Maldera, J. A., Munoz, M. W., Ernesto, J. I., Vasen, G., and Battistone, A. (2010). Behaviour of CRISP proteins during capacitation and their role in gamete interaction. In ‘Proceedings of the 11th International Symposium in Spermatology, Okinawa’. S08–06.

Colleoni S., Lagutina I., Rodriguez-Martinez H., Lazzari G., Galli C., Morrell J. M. (2011). Single layer centrifugation with Androcoll-E and HBA improve selection of the best spermatozoa for equine assisted reproduction. J. Equine Vet. Sci. in press.

Garcia, E. M., Calvete, J. J., Sanz, L., Roca, J., Martinez, E. A., and Vázquez, J. M. (2009). Distinct effects of boar seminal plasma fractions exhibiting different protein profiles on the functionality of highly diluted boar spermatozoa. Reprod. Domest. Anim. 44, 200–205.
CrossRef | CAS | PubMed |

Garner, D. L., Thomas, C. A., Gravance, C. G., Marshall, C. E., De Jarnette, J. M., and Allen, C. H. (2001). Seminal plasma addition attenuates the dilution effect of bovine sperm. Theriogenology 56, 31–40.
CrossRef | CAS | PubMed |

Hansen, C., Vermeiden, T., Vermeiden, J. P. W., Simmet, C., Day, B. C., and Feitsma, H. (2006). Comparison of FACSCount AF system, improved Neubauer hemocyometer, Corning 254 photometer, SpermVision, Ultimate and Nucleocounter SP-100 for determination of sperm concentration of boar semen. Theriogenology 66, 2188–2194.
CrossRef | CAS | PubMed |

Hollinshead, F. K., O’Brien, J. K., Maxwell, W. M. C., and Evans, G. (2004). Assessment of in vitro sperm characteristics after flow cytometric sorting of frozen–thawed bull spermatozoa. Theriogenology 62, 958–968.
CrossRef | CAS | PubMed |

Maxwell, W. M. C., de Graaf, S. P., El-Hajj Ghaouim, R., and Evans, G. (1997). Seminal plasma effects on sperm handling and female fertility. Soc. Reprod. Fertil. Suppl. 64, 13–38.

Maxwell, W. M. C., Parrilla, I., Caballero, I., Garcia, E., Roca, J., Martinez, E. A., Vazquez, J. M., and Rath, D. (2007). Retained functional integrity of bull spermatozoa after double freezing and thawing using PureSperm density gradient centrifugation. Reprod. Domest. Anim. 42, 489–494.
CrossRef | CAS |

Mocé, E., Blanch, E., Tomás, C., and Graham, J. K. (2010). Use of cholesterol in sperm cryopreservation: present moment and perspectives to future. Reprod. Domest. Anim. 45, 57–66.
CrossRef | PubMed |

Moore, A. I., Squires, E. L., and Graham, J. K. (2005). Effect of seminal plasma on the cryopreservation of equine spermatozoa. Theriogenology 63, 2372–2381.
| CAS | PubMed |

Morrell, J. M., and Rodriguez-Martinez, H. (2009). Biomimetic techniques for improving sperm quality in animal breeding: a review. The Open Andrology Journal 1, 1–9.

Morrell, J. M., and Wallgren, M. (2010). Androcoll-P-Large selects boar spermatozoa with good membrane integrity from the sperm-rich fraction of the ejaculate. Anim. Reprod. 7, 16–20.

Morrell, J. M., and Wallgren, M. (2011). Removal of bacteria from boar ejaculates by Single Layer Centrifugation can reduce the use of antibiotics in semen extenders. Anim. Reprod. Sci. 123, 64–69.
| CAS | PubMed |

Morrell, J. M., Saravia, F., Wallgren, M., van Wienen, M., and Rodriguez-Martinez, H. (2009). Selection of boar spermatozoa using centrifugation on a glycidoxypropyltrimethoxy-silane-coated silica colloid. J. Reprod. Dev. 55, 547–552.
CrossRef | PubMed |

Morrell, J. M., Mari, G., Kútvölgyi, G., Meurling, S., Mislei, B., Iacono, E., and Rodriguez-Martinez, H. (2011). Pregnancies following artificial insemination with spermatozoa from problem stallion ejaculates processed by single-layer centrifugation with Androcoll-E. Reprod. Domest. Anim. 46, 642–645.
CrossRef | PubMed |

Mortimer, D. (2000). Sperm preparation methods. J. Androl. 21, 357–366.
| CAS | PubMed |

Popwell, J. M., and Flowers, W. L. (2004). Variability in relationships between semen quality and estimates of in vivo and in vitro fertility in boars. Anim. Reprod. Sci. 81, 97–113.
CrossRef | CAS | PubMed |

Pursel, V. G., and Johnson, L. (1975). Freezing of boar spermatozoa. Fertilizing capacity with concentrated semen and a new thawing procedure. J. Anim. Sci. 40, 99–102.
| CAS | PubMed |

Roberts, K. P., Johnston, D. S., Nolan, M. A., Wooters, J. L., Waxmonsky, N. C., Piehl, L. B., Ensrud-Bowlin, K. M., and Hamilton, D. W. (2007). Structure and function of epididymal protein cysteine-rich secretory protein-1. Asian J. Androl. 9, 508–514.
CrossRef | CAS | PubMed |

Roca, J., Rodríguez, M. J., Gil, M. A., Carvajal, G., Garcia, E. M., Cuello, C., Vazquez, J. M., and Martinez, E. A. (2005). Survival and in vitro fertility of boar spermatozoa frozen in the presence of superoxide dismutase and/or catalase. J. Androl. 25, 15–24.

Samardzija, M., Karadjole, M., Getz, I., Makek, Z., Cergolj, M., and Dobranic, T. (2006b). Effects of bovine spermatozoa preparation on embryonic development in vitro. Reprod. Biol. Endocrinol. 4, 58.
CrossRef | PubMed |

Simonsson, A. (1994). ‘Näringsrekommendationer och fodermedelstabeller till svin (Nutritional recommendations and feedstuffs for swine).’ (Swedish University of Agricultural Sciences: Uppsala.)

Suarez, S. S., and Pacey, A. A. (2006). Sperm transport in the female reproductive tract. Hum. Reprod. Update 12, 1–2.

Thys, M., Vandaele, L., Morrell, J. M., Mestach, J., Van Soom, A., Hoogewijs, M., and Rodriguez-Martinez, H. (2009). In vitro fertilising capacity of frozen–thawed bull spermatozoa selected by single-layer glycidoxypropyltrimethoxysilane-coated silica colloidal centrifugation. Reprod. Domest. Anim. 44, 390–394.
CrossRef | CAS | PubMed |

Ubhayasekera, S. J. K. A., Verleyen, T., and Dutta, P. C. (2004). Evaluation of GC and GC-MS methods for the analysis of cholesterol oxidation products. Food Chem. 84, 149–157.
CrossRef | CAS |

Underwood, S. L., Bathgate, R., Gillan, L., Maxwell, W. M. C., and Evans, G. (2006). A comparison of staining media for sex-sorting cooled dairy bull spermatozoa. Reprod. Domest. Anim. 41, 378–382.

van Wienen, M., Johannisson, A., Wallgren, M., Joyce Parlevliet, J., and Morrell, J. M. (2011). Single-layer centrifugation with Androcoll-P can be scaled up to process larger volumes of boar semen. ISRN Veterinary Science , , Article ID 548385.
CrossRef |

Wongtawan, T., Saravia, F., Wallgren, M., Caballero, I., and Rodríguez-Martíneza, H. (2006). Fertility after deep intra-uterine artificial insemination of concentrated low-volume boar semen doses. Theriogenology 65, 773–787.
CrossRef | PubMed |

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