Cryopreserved sperm from non-human primates have been used for AI and IVF, but macaque sperm, like human sperm, have highly variable cryoprotection requirements depending on the individual sperm donor. This biological variability has restricted progress in developing a cryopreservation protocol that preserves sperm motility in all semen samples. This significant problem is likely to be exacerbated by significant variability in the cryoprotection requirements of sperm from genetically altered or unique individuals. The long-range goal of this proposal is to assure the preservation of valuable primate models of human disease. The principal objective is to develop a clear understanding of the fundamental cryobiology of macaque sperm. The second objective is to develop a rational and reliable method for cryopreservation of macaque sperm. The specific goals of this project are to determine the major aspects of biochemical composition and biophysical characteristics of macaque sperm, the extent of osmotic stress-induced cell damage in macaque sperm, the effects from temperature on spermatozoal responses to osmotic stress, and to determine whether cryoprotectant exposure can modulate adverse cellular properties resulting from cooling and osmotic stress. Rational development of an optimized cryopreservation protocol will then be developed based on the fundamental knowledge of macaque sperm generated from the proposed experiments. This project has a highly qualified research team comprised of cell biologists and cryobiologists from the University of California, Davis and the University of Minnesota. The project will utilize numerous techniques to accomplish these goals. Fourier-transform infrared spectroscopy will be used to assess sperm membrane phase transitions. Computerized sperm motility analysis, epifluorescence and confocal microscopy, flow cytometry, differential scanning calorimetry, and cell volumetric analysis will be used to evaluate sperm functional parameters and sublethal damage to macaque sperm. Mathematical modeling will then be used to construct optimized cryopreservation protocols.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Research Project (R01)
Project #
1R01RR016581-01A1
Application #
6544939
Study Section
National Center for Research Resources Initial Review Group (RIRG)
Program Officer
Harding, John D
Project Start
2002-06-01
Project End
2006-04-30
Budget Start
2002-06-01
Budget End
2003-04-30
Support Year
1
Fiscal Year
2002
Total Cost
$285,288
Indirect Cost
Name
University of California Davis
Department
Veterinary Sciences
Type
Schools of Veterinary Medicine
DUNS #
094878337
City
Davis
State
CA
Country
United States
Zip Code
95618
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Sumigama, Shiho; Mansell, Steven; Miller, Melissa et al. (2015) Progesterone Accelerates the Completion of Sperm Capacitation and Activates CatSper Channel in Spermatozoa from the Rhesus Macaque. Biol Reprod 93:130
Martorana, Kelly; Klooster, Katie; Meyers, Stuart (2014) Suprazero cooling rate, rather than freezing rate, determines post thaw quality of rhesus macaque sperm. Theriogenology 81:381-8
Burruel, Victoria; Klooster, Katie L; Chitwood, James et al. (2013) Oxidative damage to rhesus macaque spermatozoa results in mitotic arrest and transcript abundance changes in early embryos. Biol Reprod 89:72
Hagedorn, Mary; McCarthy, Megan; Carter, Virginia L et al. (2012) Oxidative stress in zebrafish (Danio rerio) sperm. PLoS One 7:e39397
Cole, Julie A; Meyers, Stuart A (2011) Osmotic stress stimulates phosphorylation and cellular expression of heat shock proteins in rhesus macaque sperm. J Androl 32:402-10
McCarthy, Megan J; Meyers, Stuart A (2011) Antioxidant treatment in the absence of exogenous lipids and proteins protects rhesus macaque sperm from cryopreservation-induced cell membrane damage. Theriogenology 76:168-76
Klooster, Katie L; Burruel, Victoria R; Meyers, Stuart A (2011) Loss of fertilization potential of desiccated rhesus macaque spermatozoa following prolonged storage. Cryobiology 62:161-6
McCarthy, Megan J; Baumber, Julie; Kass, Philip H et al. (2010) Osmotic stress induces oxidative cell damage to rhesus macaque spermatozoa. Biol Reprod 82:644-51
Meyers, Stuart A; Li, Ming-Wen; Enders, Allen C et al. (2009) Rhesus macaque blastocysts resulting from intracytoplasmic sperm injection of vacuum-dried spermatozoa. J Med Primatol 38:310-7

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