Response of Rhesus Sperm to Freezing: Recovery from Osmotic and Oxidative Stress

Summary

Principal Investigator: S A Meyers
Abstract: DESCRIPTION (provided by applicant): Efficient preservation of genetic resources of the rhesus monkey (Macaca mulatta) is critical to the mission of the National Center for Research Resources. This application is aimed at the development of efficient macaque sperm cryopreservation and will aid in successful propagation of germplasm through artificial insemination and in vitro fertilization programs. Non-human primates (NHPs), especially genetically unique macaques, are important models of human disease with applications to infertility and contraception, infectious diseases and vaccine development, drug and alcohol addiction, neurological disorders, and regenerative medicine. Similar to human sperm, cryopreserved sperm from NHPs show variable cryoprotection success depending on the individual sperm donor. Semen donors are often selected based on successful cryopreservation results, however, many individual males, including rhesus males in research programs, have semen quality that fails to survive cryopreservation. This is likely to be particularly true concerning NHPs generated from embryonic stem cells, somatic cell nuclear transfer, and transgenics where propagation of a specific individual's genotype is desired. This grant analyzes some of the mechanisms that may cause poor cryoperformance and focuses on methods to uniformly cryopreserve these bimodal populations. Our central hypothesis is that an understanding of cryopreservation-associated physiological responses in sperm will lead to improvements in sperm cryosurvival. Our long-term goal is to develop an understanding of crucial mechanisms involved in cryopreservation of sperm in order to optimally preserve sperm from a variety of genotypically unique and valuable NHP males. Our approach will be to investigate the mechanisms of osmotic and oxidative cellular stress at the cell physiology level which includes membrane, cell signaling, and subcellular assessment of sperm function using fluorescence microscopy, flow cytometry, cryomicroscopy, and proteomics methods. As a team with members from UC Davis, Indiana University/Purdue University, and the University of Newcastle (Australia), we are well positioned to undertake the proposed research, because of our combined experience with fundamental cryobiology and sperm cell biology.
Funding Period: ----------------2001 - ---------------2011-
more information: NIH RePORT

Top Publications

  1. ncbi Hyperactivated motility in rhesus macaque (Macaca mulatta) spermatozoa
    Julie Baumber
    Sperm Biology Laboratory, Department of Anatomy, Physiology and Cell Biology, School of Veterinary Medicine, University of California, Davis, 95616, USA
    J Androl 27:459-68. 2006
  2. pmc Oxidative damage to rhesus macaque spermatozoa results in mitotic arrest and transcript abundance changes in early embryos
    Victoria Burruel
    Department of Anatomy, Physiology, and Cell Biology, School of Veterinary Medicine, University of California Davis, Davis, California
    Biol Reprod 89:72. 2013
  3. pmc Oxidative stress in zebrafish (Danio rerio) sperm
    Mary Hagedorn
    Department of Reproductive Sciences, Smithsonian Conservation Biology Institute, Washington, District of Columbia, USA
    PLoS ONE 7:e39397. 2012
  4. pmc Suprazero cooling rate, rather than freezing rate, determines post thaw quality of rhesus macaque sperm
    Kelly Martorana
    Department of Anatomy, Physiology, and Cell Biology, School of Veterinary Medicine, University of California, Davis, CA, USA
    Theriogenology 81:381-8. 2014
  5. pmc Sperm mitochondrial integrity is not required for hyperactivated motility, zona binding, or acrosome reaction in the rhesus macaque
    Pei Hsuan Hung
    Molecular, Cellular, and Integrative Physiology, University of California, Davis, Davis, California 95616, USA
    Biol Reprod 79:367-75. 2008
  6. ncbi The macaque sperm actin cytoskeleton reorganizes in response to osmotic stress and contributes to morphological defects and decreased motility
    Liane M Correa
    School of Veterinary Medicine, Department of Anatomy, Physiology and Cell Biology, University of California, Davis, California 95616, USA
    Biol Reprod 77:942-53. 2007
  7. ncbi Changes in membrane lipid order with capacitation in rhesus macaque (Macaca mulatta) spermatozoa
    Julie Baumber
    Department of Anatomy, Physiology, and Cell Biology, School of Veterinary Medicine, University of California, Davis, California 95616, USA
    J Androl 27:578-87. 2006
  8. ncbi Biophysics of zebrafish (Danio rerio) sperm
    M Hagedorn
    Department of Reproductive Sciences, Smithsonian National Zoological Park, Washington, DC 20008, USA
    Cryobiology 58:12-9. 2009
  9. ncbi Rhesus macaque blastocysts resulting from intracytoplasmic sperm injection of vacuum-dried spermatozoa
    Stuart A Meyers
    Department of Anatomy, Physiology, and Cell Biology, University of California, Davis, CA 95616, USA
    J Med Primatol 38:310-7. 2009
  10. ncbi Osmotic stress stimulates phosphorylation and cellular expression of heat shock proteins in rhesus macaque sperm
    Julie A Cole
    Haring Hall, Department of Anatomy, Physiology, and Cell Biology, School of Veterinary Medicine, University of California, Davis, CA 95616, USA
    J Androl 32:402-10. 2011

Scientific Experts

  • S A Meyers
  • Mary Hagedorn
  • Julie Baumber
  • Megan J McCarthy
  • Katie L Klooster
  • Kelly Martorana
  • Victoria Burruel
  • Julie A Cole
  • Pei Hsuan Hung
  • Liane M Correa
  • Katie Klooster
  • James Chitwood
  • Pablo J Ross
  • Victoria R Burruel
  • Philip H Kass
  • Marion G Miller
  • Catherine A Vandevoort
  • Alysia Thomas

Detail Information

Publications13

  1. ncbi Hyperactivated motility in rhesus macaque (Macaca mulatta) spermatozoa
    Julie Baumber
    Sperm Biology Laboratory, Department of Anatomy, Physiology and Cell Biology, School of Veterinary Medicine, University of California, Davis, 95616, USA
    J Androl 27:459-68. 2006
    ..005) reduced by the addition of H-89. The results of this paper demonstrate that hyperactivation can be reliably estimated for rhesus macaque spermatozoa...
  2. pmc Oxidative damage to rhesus macaque spermatozoa results in mitotic arrest and transcript abundance changes in early embryos
    Victoria Burruel
    Department of Anatomy, Physiology, and Cell Biology, School of Veterinary Medicine, University of California Davis, Davis, California
    Biol Reprod 89:72. 2013
    ..Changes in transcript abundance in embryos destined for mitotic arrest is evident at the two-cell stage of development...
  3. pmc Oxidative stress in zebrafish (Danio rerio) sperm
    Mary Hagedorn
    Department of Reproductive Sciences, Smithsonian Conservation Biology Institute, Washington, District of Columbia, USA
    PLoS ONE 7:e39397. 2012
    ....
  4. pmc Suprazero cooling rate, rather than freezing rate, determines post thaw quality of rhesus macaque sperm
    Kelly Martorana
    Department of Anatomy, Physiology, and Cell Biology, School of Veterinary Medicine, University of California, Davis, CA, USA
    Theriogenology 81:381-8. 2014
    ..These data suggest that sperm quality seems to be more sensitive to the cooling, rather than freezing rate and highlight the role of the suprazero cooling rate in post thaw sperm quality. ..
  5. pmc Sperm mitochondrial integrity is not required for hyperactivated motility, zona binding, or acrosome reaction in the rhesus macaque
    Pei Hsuan Hung
    Molecular, Cellular, and Integrative Physiology, University of California, Davis, Davis, California 95616, USA
    Biol Reprod 79:367-75. 2008
    ....
  6. ncbi The macaque sperm actin cytoskeleton reorganizes in response to osmotic stress and contributes to morphological defects and decreased motility
    Liane M Correa
    School of Veterinary Medicine, Department of Anatomy, Physiology and Cell Biology, University of California, Davis, California 95616, USA
    Biol Reprod 77:942-53. 2007
    ..Together, our results support a mechanism in which reorganization of the actin cytoskeleton induced by osmotic stress and potentially mediated by a Rho A signaling pathway contributes to sublethal sperm flagellar and motility defects...
  7. ncbi Changes in membrane lipid order with capacitation in rhesus macaque (Macaca mulatta) spermatozoa
    Julie Baumber
    Department of Anatomy, Physiology, and Cell Biology, School of Veterinary Medicine, University of California, Davis, California 95616, USA
    J Androl 27:578-87. 2006
    ..Merocyanine may be a useful tool for investigating the role of the plasma membrane on capacitation and other cytotoxic events in macaque spermatozoa...
  8. ncbi Biophysics of zebrafish (Danio rerio) sperm
    M Hagedorn
    Department of Reproductive Sciences, Smithsonian National Zoological Park, Washington, DC 20008, USA
    Cryobiology 58:12-9. 2009
    ..Additional methods will be necessary to analyze and improve cryopreservation techniques and post-thaw fertility of zebrafish sperm. The present study is a first step to explore such techniques...
  9. ncbi Rhesus macaque blastocysts resulting from intracytoplasmic sperm injection of vacuum-dried spermatozoa
    Stuart A Meyers
    Department of Anatomy, Physiology, and Cell Biology, University of California, Davis, CA 95616, USA
    J Med Primatol 38:310-7. 2009
    ..In this study, we examined the feasibility and efficiency of intracytoplasmic sperm injection using vacuum-dried rhesus macaque sperm in CZB medium supplemented with 10% fetal bovine serum...
  10. ncbi Osmotic stress stimulates phosphorylation and cellular expression of heat shock proteins in rhesus macaque sperm
    Julie A Cole
    Haring Hall, Department of Anatomy, Physiology, and Cell Biology, School of Veterinary Medicine, University of California, Davis, CA 95616, USA
    J Androl 32:402-10. 2011
    ..Taken together, these results suggest a differential role for HSP 70 and HSP 90 during osmotic stress conditions in rhesus macaque sperm...
  11. ncbi Loss of fertilization potential of desiccated rhesus macaque spermatozoa following prolonged storage
    Katie L Klooster
    Department of Anatomy, Physiology, and Cell Biology, School of Veterinary Medicine, University of California Davis, CA, USA
    Cryobiology 62:161-6. 2011
    ..This study demonstrates that while the vacuum dried spermatozoa were immotile and had compromised plasma membrane integrity, they were capable of fertilization using ICSI and could support embryo development to the morula stage...
  12. pmc Antioxidant treatment in the absence of exogenous lipids and proteins protects rhesus macaque sperm from cryopreservation-induced cell membrane damage
    Megan J McCarthy
    Department of Anatomy, Physiology, and Cell Biology, School of Veterinary Medicine, University of California, Davis, CA, USA
    Theriogenology 76:168-76. 2011
    ....
  13. pmc Osmotic stress induces oxidative cell damage to rhesus macaque spermatozoa
    Megan J McCarthy
    Departments of Anatomy, Physiology, and Cell Biology, School of Veterinary Medicine, University of California, Davis, CA, USA
    Biol Reprod 82:644-51. 2010
    ..These results clearly indicate that osmotic stress causes oxidative stress in rhesus macaque spermatozoa, which strongly supports the hypothesis that cryopreservation-induced osmotic stress may lead to oxidative cell damage...