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Characterization of Oxytocin Receptors in Corpus Luteum Endothelial Cells

Characterization of Oxytocin Receptors in Corpus Luteum Endothelial Cells. Katy McHenry & Dr. Fredrick Stormshak Department of Animal Sciences Oregon State University. Background. Bovine Estrous Cycle = 21 days One egg matures Released as an oocyte

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Characterization of Oxytocin Receptors in Corpus Luteum Endothelial Cells

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  1. Characterization of Oxytocin Receptors in Corpus Luteum Endothelial Cells Katy McHenry & Dr. Fredrick Stormshak Department of Animal Sciences Oregon State University

  2. Background • Bovine Estrous Cycle = 21 days • One egg matures • Released as an oocyte • Granulosa cells of the empty follicle turn into the corpus luteum • Angiogenesis • Endocrine Gland

  3. Oxytocin • Smooth muscle contraction in uterus during childbirth • Roll in maternal instinct and building relationships? Stimulation of milk letdown

  4. Why??

  5. Hypothesis Bovine luteal oxytocin acts locally to stimulate the proliferation and migration of endothelial cells to form the vascular system of the developing corpus luteum.

  6. 1.Block oxytocin receptors Importance Corpus Luteum can not form Progesterone 2. Tumor angiogenesis and growth

  7. Procedure 1. Isolate the endothelial cells 2. Measure the binding of oxytocin to endothelial cell oxytocin receptors

  8. Isolating Endothelial Cells Remove corpus luteum eight days post-estrus • Check cows for estrus using vasectomized bull

  9. Dissociate corpus luteum using collagenase and DNAse • Tosyl-activated magnetic beads • Couple with lectin BS-1 • Add dissociated tissue and separate endothelial cells

  10. radioreceptor Assay for oxytocin binding [ H]-Oxytocin & 200 fold excess unlabeled Oxytocin • [ H]-Oxytocin 3 3

  11. Radioreceptor assay for oxytocin binding

  12. Results

  13. Results

  14. New Theory • Pericyte Cells • Stem Cells Smooth Muscle

  15. Results

  16. Summary • Oxytocin showed slight binding to receptors on endothelial cells in the corpus luteum. • Hypothesis was not strongly supported • Large amount of oxytocin binding to receptors on non-endothelial luteal cells • Possibly pericyte cells?

  17. Thank you • Dr. Fred Stormshak • Oregon State University, Department of Animal Sciences • Oregon State University, Honors College • Dr. Kevin Ahern • The Howard Hughes Medical Institute

  18. Questions?

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