Daigneault Laboratory

Focus - Environmental Influence on Postejaculatory Sperm Function Through Embryo Development

The laboratory specializes in utilizing large animal models to address emerging and existing reasons for infertility in animals and humans. We primarily use bovine gametes and embryos to address idiopathic subfertility that also serve as models for human embryo development. Existing projects in the laboratory equally span male and female reproductive biology with a concentric focus on basic functions that drive early embryo development for applied and translational agriculture and biomedical sciences. Assisted reproductive technologies are a large component of research (IVF, ET, AI, vitrification) spanning multiple species (bovine, equine, ovine, porcine). Complementary techniques include the use of CRISPR/Cas technologies (zygotic injection, electroporation) for gene editing to identify genomic requirements of early embryo development and pregnancy recognition including paternal contributions to blastocyst formation. In complement, the laboratory models gamete and embryo environmental interactions with a focus on contaminants and toxicants that alter sperm function and embryo development both structurally and at the genome/epigenomic levels. We also optimize and design semen extenders for multiple purposes in different species for commercial and research applications.

Existing Projects

• Functional characterization of genes required for early blastocyst formation and trophoblast function in cattle
• Roles of nuclear and mitochondrial proteins in sperm function and fertilization
• Impact of environmental exposure to extra-gonadal sperm on fertilization potential and embryo development
• Optimization of techniques and conditions to improve storage time in cooled stallion semen
• Development of customized bull semen extenders for specific applications (sex-sorting, long-term storage)
• Characterization of Florida Native Sheep seminal parameters

Gene Editing - CRISPR technologies

Infertility and embryonic failure are highly evident in both humans and cattle. The laboratory is building from previously published work to demonstrate conserved function in pluripotent genes as master regulators of embryonic development in cattle and humans. Delivery of CRISPR/Cas9 components into bovine zygotes is achieved to generate gene-edited embryos intended to understand basic biology and the genetic architecture required for successful establishment and maintenance of pregnancy.

Environmental Toxicology

Paternal exposure to environmental toxicants is a highly understudied area of research in large animals and humans. Our works aim to identify and determine consequences of environmental exposure of common contaminants to post-ejaculatory sperm function and subsequent embryo development. Using a bovine model and embryo continuum, sperm are exposed to physiological and environmentally relevant levels of toxicants to understand consequences on sperm function including motility, fertilization, and subsequent embryo development.

Semen Extenders/Diluents

Our laboratory optimizes and designs semen extenders primarily in bovine and equine for specific needs including transportation at ambient temperature, sex-sorting, increased velocity for timed-AI protocols, and prolonged maintenance of frozen-thawed sperm. Extenders are validated for compatibility in the laboratory using Computer-Assisted Sperm Analyses and submitted for Patent approval towards industry and research applications.

Interdisciplinary Involvement

• Reproductive and Perinatal Biology Research Program
• Animal Molecular & Cellular Biology
• UF Equine Sciences

Courses Taught

• Reproductive Physiology and Endocrinology in Domestic Animals – ANS3319C
• Applied Animal Reproduction – ANS4320C & ANS6312C