Masters Theses

Date of Award

12-2002

Degree Type

Thesis

Degree Name

Master of Science

Major

Animal Science

Major Professor

F. Neal Schrick

Committee Members

Lannett Edwards, Patty Tithof, Jay Whelan

Abstract

Two studies were performed to determine effects of prostaglandin F2 α (PGF2 α) on continued embryonic development of pre-compacted (in vitro-produced) and compacted (in vitro-derived) bovine embryos. In Experiment 1, pre-compacted (16-32 cell) in vitro-produced embryos were randomly assigned to receive either: 1) control (KSOM-PVA; n= 168); 2) PGF-1 (1 ng/mL PGF in KSOM-PVA; n= 143); 3) PGF-10 (10 ng/mL PGF in KSOM-PVA; n= 168); 4) PGF-100 (100 ng/mL PGF in KSOM-PVA; n= 136), or 5) PGE-5 (5 ng/mL PGE2 in KSOM-PVA; n= 62). Following four days of culture (5.5% CO2, 7% O2, and 87.5% N2 at 38.5°C) in assigned treatments, development of embryos was determined by experienced technicians uninformed of treatments. Continued development of in vitro-produced 16- to 32-cell embryos (pre-compacted) to blastocyst was reduced by addition of PGF in culture media (PGF-1, 30.4%; PGF-10, 41.4%; PGF-100, 33.3%, and control, 51.8%; P=0.002). In addition, culture of embryos in medium containing 1 ng/mL of PGF had a more detrimental effect than addition of 10 ng/mL of PGF (P<0.05). Furthermore, blastocyst development did not differ between control and PGE2 treatments (P>0.10).

In Experiment 2, compacted (in vitro-derived) glycerol-frozen embryos were thawed and then allowed to regain normal morphology in holding medium for 30 min. Embryos were then sorted by stage of development and quality according with IETS guidelines for classification of bovine embryos. Compacted morula in the quality score of 1, 2 or 3 (n= 436) were randomly assigned to one of four treatments: 1) control (KSOM-PVA; n= 110); 2) PGF-0.1 (0.1 ng/mL PGF in KSOM-PVA; n=108); 3) PGF-1 (1 ng/mL PGF in KSOM-PVA; n=109); 4) PGF-10 (10 ng/mL PGF in KSOM-PVA; n=109). After 24 h in culture (5.5% CO2, 7% O2, and 87.5% N2 at 38.5°C), embryos were washed and placed in KSOM containing 0.5% BSA without PGF for an additional 48 h. Embryo development was evaluated by experienced technicians uninformed of treatments. Continued development of in vitro-derived compacted morula to blastocyst was not affected by addition of PGF to the culture medium (control, 68.1%; PGF-0.1, 58.9%; PGF-1, 61%, and PGF-10, 60%; P>010). However, hatching rates of embryos cultured with 0.1, 1 and 10 ng/mL PGF were significantly lower than controls (24, 29.1, 24.5, and 44.5, respectively; P=0.05). No significant differences in hatching rates between doses of PGF were observed. In conclusion, these studies suggest a direct negative effect of PGF on continued embryonic development of pre-compacted and compacted bovine embryos. Responses to PGF of pre-compacted (decreased blastocyst development) and compacted (decreased hatching rates) embryos may involve different mechanisms of action of PGF depending on stage of development. In compacted embryos, PGF may act through inhibiting completion of blastocoel formation and hatching; whereas, PGF may have more dramatic effects by altering compaction in pre-compacted embryos.

Files over 3MB may be slow to open. For best results, right-click and select "save as..."

Share

COinS