Towards The Dry Preservation of Feline Sperm: The Effect Of Dehydration On DNA, Centrosomal Function, And Embryo Development

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Patrick, J. (2016). Towards The Dry Preservation of Feline Sperm: The Effect Of Dehydration On DNA, Centrosomal Function, And Embryo Development. Unc Charlotte Electronic Theses And Dissertations.
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Abstract

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.

Details
Author

Patrick, Jennifer

Title

Towards The Dry Preservation of Feline Sperm: The Effect Of Dehydration On DNA, Centrosomal Function, And Embryo Development

Physical Description

1 online resource (103 pages) : PDF

Date

2016

Degree Granting Institution

University of North Carolina at Charlotte

Abstract

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.

Genre

doctoral dissertations

Subjects--Topics

Biology

Degree

Ph.D.

Keywords

Centrosome
Damage
Dehydration
DNA
Feline
Sperm

Subject Area

Biology

Advisor(s)

Elliott, Gloria

Committee Members

Comizzoli, Pierre
Huet, Yvette
Yan, Shan
Trammell, Susan

Degree Note

Thesis (Ph.D.)--University of North Carolina at Charlotte, 2016.

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Rights Holder Information

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Identifier

Patrick_uncc_0694D_11058

Permalink

http://hdl.handle.net/20.500.13093/etd:533