Development and de novo protein synthetic activity of bovine embryos produced in vitro in different culture systems.
In vitro matured (IVM) and fertilized (IVF) putative Day 1 zygotes (Day 0 = IVF) were allocated randomly to culture in formulations based on Synthetic Oviduct Fluid (SOF) medium and identified on the basis of their contrasting principal supplements, which were 10% v/v steer serum (SS; n = 558) or 4 mg/mL crystalline BSA (SBSA; n = 531) or 3 mg/mL polyvinyl alcohol (SPVA; n = 607) in 9 replicates. SBSA and SPVA also contained 10 microg/mL non-essential amino acids, while the former was further supplemented with 20 microL/mL essential amino acids and the latter with 0.5 mmol/L sodium citrate and 5 ng/mL epidermal growth factor. Zygotes were cultured in 20 microL drops (4 zygotes per drop) until Day 8 in an atmosphere of 5% CO2, 5% O2 and 90% N2 at 39 degrees C and droplets were renewed every 48 hours. The incidence of zygote cleavage was lower (P < 0.05) in SS (mean +/- SEM = 61 +/- 3%) than in SBSA (76 +/- 3%) but not in SPVA (72 +/- 4%) up to Day 3. The SPVA generated a lower yield of blastocysts on Day 7 (12 +/- 2%; P < 0.001) and by Day 8 (21 +/- 4%; P < 0.01) than did SS (33 +/- 3%; 40 +/- 3%) and SBSA (30 +/- 3%; 37 +/- 4%). Cell numbers (n) and diameters (d) of blastocysts on Day 8 were greater (P < 0.001; Replicates 1 to 5) in embryos from SBSA (n, 156 +/- 9; d, 203 +/- 4 microm) than in those from SS (n, 81 +/- 4; d, 177 +/- 3 microm) and SPVA (n, 76 +/- 5; d, 167 +/- 3 microm). Embryos produced in SS incorporated less 3H-phenylalanine into PCA-precipitable protein (replicates 6 to 9; log10 dpm = 3.03 +/- 0.04) than did embryos cultured in SBSA (3.21 +/- 0.03; P < 0.001) or in SPVA (3.14 +/- 0.03; NS). In conclusion, blastocyst yield was poor in SPVA, but the embryos had metabolic activities similar to those of embryos produced in SBSA. Blastocyst yields from SS were not compromised but their capacity for de novo protein synthesis was reduced significantly.
Scottish Agricultural College, Animal Biology Division, Aberdeen, UK. mkuran@omu.edu.tr
M
M
Kuran
J J
JJ
Robinson
M E
ME
Staines
T G
TG
McEvoy
D000818:Animals; D001755:Blastocyst / Q000502:physiology; D001757:Blastomeres / Q000378:metabolism; D002417:Cattle / Q000196:embryology* / Q000502:physiology; D002951:Citrates / Q000502:physiology; D016895:Culture Media, Serum-Free; D005314:Embryonic and Fetal Development / Q000502:physiology; D004815:Epidermal Growth Factor / Q000502:physiology; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary*; D006651:Histocytochemistry; D008297:Male; D010649:Phenylalanine / Q000737:chemistry; D011142:Polyvinyl Alcohol / Q000494:pharmacology; D011247:Pregnancy; D014176:Protein Biosynthesis; D011506:Proteins / Q000032:analysis; D011897:Random Allocation; D012710:Serum Albumin, Bovine / Q000502:physiology; D000077559:Sodium Citrate; D015053:Zygote / Q000254:growth & development / Q000378:metabolism
D002951:Citrates; D016895:Culture Media, Serum-Free; D011506:Proteins; D000077559:Sodium Citrate; D012710:Serum Albumin, Bovine; D010649:Phenylalanine; D004815:Epidermal Growth Factor; D011142:Polyvinyl Alcohol