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United States Department of Agriculture

Agricultural Research Service

Title: Cryopreservation of Flow Cytometrically Sorted Boar Sperm: Effects on in Vivo Embryo Development

Authors
item Johnson, L - COLLABORATOR, ARS
item Guthrie, Howard
item Fiser, P - OTTAWA, ONTARIO, CANADA
item Maxwell, W.M.C. - UNIVERSITY OF SYDNEY
item Welch, Glenn
item Garrett, Wesley

Submitted to: Joint Abstracts of the American Dairy Science and Society of Animal Science
Publication Type: Abstract Only
Publication Acceptance Date: May 8, 2000
Publication Date: N/A

Technical Abstract: Aliquots of neat semen from mature boars was diluted and stained with Hoechst 33342, incubated at 35 C and sorted at room temperature (SORT) into TEST-Yolk (20%) with out regard to separation of the X and Y sperm populations. Sorted samples were frozen in 3% glycerol and control semen in 11% lactose(UNSORT), respectively. Ovulation was controlled in 11 gilts by an altrenogest-PMSG-hCG regime. Gilts were laparotomized at 44 hr post-hCG and inseminated with 200,000 SORT or UNSORT into the isthmus of respective oviducts. Gilts were slaughtered and embryos recovered at 43 hr post insemination. The embryos were fixed in 4% paraformaldehyde, and stained with Texas Red-x phalloidin and Hoechst 33342. Confocal laser microscopy was used to visualize actin cytoskeleton and chromatin configuration. A total of 287 oocytes and embryos were evaluated. Cryopreservation and the subsequent thawing of the SORT sperm had a negative impact on fertilization nand embryo development compared to UNSORT sperm (at P level less than .05) as indicated by the incidence of sperm penetrated ova and total embryos (41.2 vs 96.5%), normal embryos without fragmentation or polyspermy (4.36 vs10.5/oviduct), incidence of normal embryos (55.4 vs 83.2%), and the proportion of 5-9 cell embryos of normal embryos (2.7 vs 21.4%); polyspermy (9.7%) and fragmentation (12.1%) were not different. These results demonstrate the combined negative impact of flow cytometric sorting and the sperm freezing and thawing process on fertilization rate and embryo development in the pig.

Last Modified: 8/30/2014
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