Towards The Dry Preservation of Feline Sperm: The Effect Of Dehydration On DNA, Centrosomal Function, And Embryo Development
1 online resource (103 pages) : PDF
University of North Carolina at Charlotte
The current gold standard for gamete preservation is cryogenic storage. Dry preservation is an attractive alternative, eliminating the need for refrigeration, reducing storage maintenance costs, and providing logistical flexibility for shipping. The current study focuses on understanding the injury during dehydration of feline epididymal sperm. This animal model serves as a closer model to humans than a rodent species because of centrosomal inheritance from the sperm. Drying to end moisture levels of 0.16 gH20/gDW did not significantly increase sperm nuclear DNA damage compared to control samples. Large sperm aster formation 5 hours post insemination (hpi) was significantly lower (p<0.05) in the dried group (5.6%) compared to control groups (14.6%), indicating a level of centrosomal damage during dehydration. No difference in the overall level of sperm DNA decondensation at 5hpi was noted. Analysis of embryo development post dehydration showed a significant decrease (p<0.01) in development to the blastocyst stage in the dehydrated group (6.4%) compared to the fresh group (15.1%). The mRNA levels, however, in developed blastocysts from dried sperm were not significantly different from the control group, suggesting equal fetal development capacity. Together, these data add to the current understanding of cellular damage during dehydration of sperm and supports the feasibility of anhydrous preservation. Future studies to evaluate specific embryonic gene activation patterns and centrosomal replacement post-dehydration could yield further advances.
Comizzoli, PierreHuet, YvetteYan, ShanTrammell, Susan
Thesis (Ph.D.)--University of North Carolina at Charlotte, 2016.
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