Title page for ETD etd-03242004-175322


Type of Document Master's Thesis
Author Klumpp, Angela Marie
Author's Email Address aklump2@lsu.edu
URN etd-03242004-175322
Title The Effect of Holding Bovine Oocytes in Follicular Fluid on Subsequent Fertilization and Embryonic Development
Degree Master of Science (M.S.)
Department Animal Science (Animal, Dairy, & Poultry Sciences)
Advisory Committee
Advisor Name Title
Robert Godke Committee Chair
Dale Paccamonti Committee Member
Stanley Leibo Committee Member
Keywords
  • bovine
  • in vitro fertilization
  • in vitro maturation
  • follicular fluid
Date of Defense 2003-12-12
Availability unrestricted
Abstract
The objective of Experiment 1 was to determine the effect of bovine follicular fluid (bFF) on nuclear maturation. Treatment A (Control) oocytes were stained with Hoechst-33342 immediately after aspiration from follicles, whereas, oocytes in Treatment B were held in bFF for 12 hours at 38¢ªC and then stained to determine nuclear status. No significant difference was detected between treatment groups. Results indicate that bFF inhibits resumption of meiosis. The objective of Experiment 2 was to determine the effect of bFF on embryonic development. Oocytes in Treatment A (Control) were placed into in vitro maturation (IVM) for 22 hours followed by in vitro fertilization (IVF). Oocytes in Treatment B were held in bFF for 12 hours at 22¢ªC, followed by IVM and then subjected to IVF. Significantly more (P<0.0001) oocytes cleaved, developed into blastocysts and hatched in Treatment A compared with Treatment B. Results indicate that a 12-hour holding period in bFF does not promote normal embryonic development. The objective of Experiment 3 was to determine the effect of decreased time and concentration of bFF on embryonic development. Treatment A (Control) oocytes were placed into IVM followed by IVF. Oocytes in Treatment B were held in bFF, oocytes in Treatment C were held in Lactated Ringer¡¯s Solution (LRS) and oocytes in Treatment D were held in a combination of bFF and LRS for 6 hours at 22¢ªC, followed by IVM then by IVF. No significant difference was detected between Treatments A and B when analyzing cleavage, blastocyst formation and hatching rates. However, significantly fewer (P<0.0001) embryos reached these stages of development in Treatments C and D. Nevertheless, there were significantly more embryos that developed to the blastocyst stage in Treatment D compared with Treatment C. Decreasing the amount of time that oocytes were held in bFF proved to be beneficial in supporting in vitro embryo production (IVP). These findings could be advantageous when attempting to rescue valuable gametes from deceased females.
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