The objective of this project is to devise technical protocols for in vitro maturation (IVM) and cryopreservation of non-human primate oocytes using rhesus monkeys. At present, IVM success in primates is very poor compared with laboratory species such as cattle. Different culture methods for supporting primate IVM will be tested to find an optimal protocol. IVM protocols will be empirically determined by testing the addition of specific amino acids, energy substrates and growth factors, which has proved successful with oocytes of other laboratory species. A key part of this approach will be use of protein-free culture media to avoid unknown factors associated with serum or serum albumin preparations, leading to development of more reproducible protocols. In a parallel study, the ability of oocytes to survive a range of osmotic shocks, cooling rates and types of cryoprotective agents will be measured. This information will be used to construct new cryopreservation protocols. The success of both IVM and cryopreservation protocols will be measured by morphological and developmental criteria, including determination of the integrity of metaphase II spindle structure, cortical granules and zonae pellucidae; examination of the organization of oocyte cytoplasm (e.g., mitochondrial distribution) using confocal and multiphoton microscopy; and assessment of viability of oocytes by their ability to undergo fertilization and embryo development, using standard protocols. Embryo development will be further evaluated by timelapse videomicrography and by embryo transfers to validate normality. Then, cryopreserved oocytes will be used to test the ability of the optimized IVM protocol to support maturation of stressed oocytes. An effective system of primate oocyte cryopreservation and IVM would provide a means for producing and making available large numbers of primate embryos for other studies on early development and would also assist primate breeding and conservation efforts.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Research Project (R01)
Project #
5R01RR013439-04
Application #
6394700
Study Section
National Center for Research Resources Initial Review Group (RIRG)
Program Officer
Harding, John D
Project Start
1998-08-01
Project End
2002-07-31
Budget Start
2001-08-01
Budget End
2002-07-31
Support Year
4
Fiscal Year
2001
Total Cost
$317,943
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
Puttabyatappa, Muraly; Brogan, Rebecca S; Vandevoort, Catherine A et al. (2013) EGF-like ligands mediate progesterone's anti-apoptotic action on macaque granulosa cells. Biol Reprod 88:18
Mtango, Namdori R; Sutovsky, Miriam; Vandevoort, Catherine A et al. (2012) Essential role of ubiquitin C-terminal hydrolases UCHL1 and UCHL3 in mammalian oocyte maturation. J Cell Physiol 227:2022-9
Mtango, Namdori R; VandeVoort, Catherine A; Latham, Keith E (2011) Ontological aspects of pluripotency and stemness gene expression pattern in the rhesus monkey. Gene Expr Patterns 11:285-98
Tollner, Theodore L; Dong, Qiaoxiang; VandeVoort, Catherine A (2011) Frozen-thawed rhesus sperm retain normal morphology and highly progressive motility but exhibit sharply reduced efficiency in penetrating cervical mucus and hyaluronic acid gel. Cryobiology 62:15-21
Dong, Qiao-Xiang; Rodenburg, Sarah E; Hill, Dana et al. (2011) The role of low-density lipoprotein (LDL) and high-density lipoprotein (HDL) in comparison with whole egg yolk for sperm cryopreservation in rhesus monkeys. Asian J Androl 13:459-64
Duffy, Diane M; VandeVoort, Catherine A (2011) Maturation and fertilization of nonhuman primate oocytes are compromised by oral administration of a cyclooxygenase-2 inhibitor. Fertil Steril 95:1256-60
Brogan, Rebecca S; MacGibeny, Margaret; Mix, Scott et al. (2011) Dynamics of intra-follicular glucose during luteinization of macaque ovarian follicles. Mol Cell Endocrinol 332:189-95
Vandevoort, Catherine A; Mtango, Namdori R; Latham, Keith E et al. (2011) Primate preimplantation embryo is a target for relaxin during early pregnancy. Fertil Steril 96:203-7
Dong, Qiaoxiang; Tollner, Theodore L; Rodenburg, Sarah E et al. (2010) Antioxidants, Oxyrase, and mitochondrial uncoupler 2,4-dinitrophenol improved postthaw survival of rhesus monkey sperm from ejaculates with low cryosurvival. Fertil Steril 94:2359-61
Duffy, Diane M; McGinnis, Lynda K; Vandevoort, Catherine A et al. (2010) Mammalian oocytes are targets for prostaglandin E2 (PGE2) action. Reprod Biol Endocrinol 8:131

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