The Candidate will receive training while producing scholarly knowledge about felid oocyte cryobiology. Since many rare cat genotypes reproduce poorly, gamete cryopreservation combined with in vitro embryo production and transfer in recipient females can help to reproduce and manage these populations. Objectives are to: 1) investigate membrane biophysical properties of the oocyte;2) characterize and mitigate cryoinjuries caused by two-step freezing;and 3) apply a newly developed oocyte cryopreservation protocol to rare felid biomedical models to assist in population management. Because of the potential of this technology for 'rescuing'gametes from dying animals, the focus will be on the immature oocyte. Impact of cryoprotectant exposure, direct cooling and holding at a subzero temperature followed by direct plunging into liquid nitrogen will be characterized. Evaluation targets include biophysical membrane properties, intracellular ice formation, integrity of the cytoskeleton, cytoplasmic membrane and interactions with cumulus cells, and eventually developmental competence in vitro and in vivo. Findings will be applied to felid models while taking advantage of unique access to selected wild felid species to more rapidly understand mechanisms. This fundamental and comparative approach provides a multidisciplinary training opportunity that will allow the Candidate to advance as a physiologist with expertise in biophysics, cryobiology and in vitro culture of oocytes. The research benefit will be a thorough understanding of the biophysical properties of the immature cat oocyte and the factors regulating its ability to survive cooling-freezing-thawing and to develop into a viable embryo and offspring. The practical benefit will be another tool for addressing the always difficult task of managing small populations of reproductively compromised felids. Relevance to public health: Animal models are essential to studying mechanisms pertinent to understanding and improving the human condition through biomedical research. This multidisciplinary approach will provide new oocyte cryopreservation protocols useful to the management of cat colonies used as biomedical models for human diseases.
