1. bookVolume 12 (2012): Issue 3 (July 2012)
Journal Details
License
Format
Journal
eISSN
2300-8733
ISSN
1642-3402
First Published
25 Nov 2011
Publication timeframe
4 times per year
Languages
English
access type Open Access

Relationship Between Apoptotic-Like Changes in Stored Boar Semen and DNA Fragmentation in Preimplantation Embryos

Published Online: 24 Jul 2012
Volume & Issue: Volume 12 (2012) - Issue 3 (July 2012)
Page range: 357 - 366
Journal Details
License
Format
Journal
eISSN
2300-8733
ISSN
1642-3402
First Published
25 Nov 2011
Publication timeframe
4 times per year
Languages
English
Relationship Between Apoptotic-Like Changes in Stored Boar Semen and DNA Fragmentation in Preimplantation Embryos

The aim of this experiment was to study the relationship between apoptotic-like changes in spermatozoa and DNA fragmentation in embryos obtained after insemination with fresh and stored semen. The ejaculates collected from three boars (five ejaculates from the same boar) were extended in Biosolwens Plus extender and stored for five days at 15-17°C. Semen, both fresh (Day 0) and stored (Day 5) used for insemination was analysed to detect apoptotic-like changes using fluorescence method: an assay to assess early changes in the membrane integrity of the sperm using the YO-PRO-1 fluorophore. After 5.5 days of insemination embryos were flushed out of the uterus and DNA fragmentation using TUNEL was analysed. In the fresh semen an average of 2.7, 3.7 and 6.2% of apoptotic sperm was observed in boar nos. 1, 2 and 3, respectively. After five days of storage the percentage of apoptotic sperm significantly increased up to 8.0, 15.7 and 23.2% in each analysed boar. The TUNEL index was 7.1% in the morphologically normal expanded blastocysts obtained after insemination with stored semen, and approximately 1.7% after insemination with fresh semen. A greater number of degenerated embryos and higher incidence of DNA fragmentation in the morphologically normal blastocysts were observed after insemination with stored semen which consists of higher percentage of apoptotic sperm compared to results from insemination with fresh semen.

Keywords

Anzar M., Liwei H., Buhr M. M., Kroetsch T. G., Pauls K. P. (2002). Sperm apoptosis in fresh and cryopreserved bull semen detected by flow cytometry and its relationship with fertility. Biol. Reprod., 66: 354-360.Search in Google Scholar

Brison D. R., Schultz R. M. (1997). Apoptosis during mouse blastocyst formation: evidence for a role for survival factors including transforming growth factor. Biol. Reprod., 56: 1088-1096.Search in Google Scholar

Bryła M., Trzcińska M., Wieczorek J. (2009). Analysis of in vivo- and in vitro-derived pig expanded blastocysts based on DNA fragmentation. Anim. Sci. Pap. Rep., 27: 59-68.Search in Google Scholar

Chen Z., Hauser R., Trbovich A. M., Shifren J. L., Dorer D. J., Godfrey-Bailey L., Singh N. P. (2006). The relationship between human semen characteristics and sperm apoptosis: a pilot study. J. Androl., 27: 112-120.Search in Google Scholar

Evenson D. P. (1999). Flow cytometry assays of male fertility. Method. Cell Biol., 33: 401-410.Search in Google Scholar

Gjørret J. O., Knijn H. M., Dieleman S. J., Avery B., Larsson L-I., Maddox-Hyttel P. (2003). Chronology of apoptosis in bovine embryos produced in vivo and in vitro. Biol. Reprod., 69: 1193-1200.Search in Google Scholar

Hao Y., Lai L., Mao J., Im G. S., Bonk A., Prather R. S. (2003). Apoptosis and in vitro development of preimplantation porcine embryos derived in vitro or by nuclear transfer. Biol. Reprod., 69: 501-507.Search in Google Scholar

Idziorek T., Estaquier J., De Bels F., Ameisen J. C. (1995). YOPRO-1 permits cytofluorometric analysis of programmed cell death (apoptosis) without interfering with cell viability. J. Immunol. Methods, 185: 249-258.Search in Google Scholar

Long C. R., Dobrinsky J. R., Garrett W. M., Johnson L. A. (1998). Dual labelling of the cytoskeleton and DNA strand breaks in porcine embryos produced in vivo and in vitro. Mol. Reprod. Dev., 51: 59-65.Search in Google Scholar

Marchetti C., Obert G., Deffosez A., Formstecher P., Marchetti P. (2002). Study of mitochondrial membrane potential, reactive oxygen species, DNA fragmentation and cell viability by flow cytometry in human sperm. Hum. Reprod., 17: 1257-1265.Search in Google Scholar

Martin G., Sabido O., Durand P., Levy R. (2004). Cryopreservation induces an apoptosis-like mechanism in bull sperm. Biol. Reprod., 71: 28-37.Search in Google Scholar

Mateusen B., Van Soom A., Maes D. G., Donnay I., Duchateau L., Lequarre A. S. (2005). Porcine embryo development and fragmentation and their relation to apoptotic markers: a cinematographic and confocal laser scanning microscopic study. Reproduction, 129: 443-452.Search in Google Scholar

Neuber E., Luetjens C. M., Chan A. W., Schatten G. P. (2002). Analysis of DNA fragmentation of in vitro cultured bovine blastocysts using TUNEL. Theriogenology, 57: 2193-2202.Search in Google Scholar

Pomar R. F. J., Teerds K. J., Kidson A., Colenbrander B., Tharasant T., Aguilar B., Roelen B. A. J. (2005). Differences in the incidence of apoptosis between in vivo and in vitro produced blastocysts of farm animal species: a comparative study. Theriogenology, 63: 2254-2268.Search in Google Scholar

Said T., Agarwal A., Grunewald S., Rasch M., Baumann T., Kriegel C., Li L., Glander H. J., Thomas A. J. Jr, Paasch U. (2006). Selection of nonapoptotic spermatozoa as a new tool for enhancing assisted reproduction outcomes: an in vitro model. Biol. Reprod., 74: 530-537.Search in Google Scholar

Szułdrzyńska J., Smorąg Z. (1980). Morphology and rate of in vitro development of mouse embryos frozen in liquid nitrogen (in Polish). Rocz. Nauk. Zoot., 7: 19-30.Search in Google Scholar

Taylor S. L., Weng S. L., Fox P., Duran E. H., Morshedi M. S., Oehninger S., Beebe S. J. (2004). Somatic cell apoptosis markers and pathways in human ejaculated sperm: potential utility as indicators of sperm quality. Mol. Hum. Reprod., 10: 825-834.Search in Google Scholar

Trzcińska M., Bryła M., Bochenek M., Słomski R., Smorąg Z. (2009). Assessment of plasma membrane and chromatin structure of sperm from transgenic and non-transgenic boars. Theriogenology, 72: 1141-1147.Search in Google Scholar

Trzcińska M., Bryła M., Smorąg Z. (2011). Apoptotic-like changes in the spermatozoa of fresh and stored boar semen and the quality of embryos produced in vivo. Anim. Reprod. Sci., 124: 90-97.Search in Google Scholar

Recommended articles from Trend MD

Plan your remote conference with Sciendo