Gene expression and developmental competence of bovine embryos produced in vitro with different serum concentrations.
The objective of this study was to determine the developmental rates and relative abundance of Hsp 70.1 and Glut-1 transcripts in in vivo- and in vitro-produced (IVP) bovine embryos in media supplemented with bovine serum albumin (BSA) or different oestrous cow serum concentrations. In experiment 1, in vitro maturation and culture media were supplemented with 0.4% BSA or 1, 5, 10 or 20% of oestrous cow serum (ECS). The analysis of the expression of Hsp 70.1 and Glut-1 was carried out in individual days 7 and 8 embryos by a semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) assay. In experiment 2, in vivo-produced morulae were collected on day 7 of the oestrous cycle and employed for the comparison of the relative abundances of Hsp 70.1 and Glut-1 transcripts with IVP morulae produced using two protein sources (10% ECS or 0.4% BSA). No differences were observed in cleavage rate among groups, but blastocyst formation (27%) and hatching rates (78%) were significantly higher in IVP embryos produced with 20% ECS than the other groups (p<0.05). No significant differences were observed in the relative abundances of Hsp 70.1 and Glut-1 mRNA in days 7 and 8 blastocysts expanded blastocysts between groups. The abundances of mRNA for those genes were similar between IVP and in vivo-produced morulae. In spite of the alterations observed in embryonic development, the presence of serum at distinct concentrations did not appear to alter the relative abundance profiles of Hsp 70.1 and Glut-1 compared with controls or the BSA supplementation to the IVP media.
Oliveira
A T D
AT
Embryology and Biotechnology of Reproduction Laboratory, Faculty of Veterinary Medicine, Universidade Federal do Rio Grande do Sul, Av Bento Goncalves, Porto Alegre RS, Brazil.
Lopes
R F F
RF
Rodrigues
J L
JL
D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000378:metabolism; D002417:Cattle / Q000196:embryology* / Q000502:physiology; D003470:Culture Media / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D046149:Embryo Culture Techniques / Q000379:methods / Q000662:veterinary*; D005260:Female; D018507:Gene Expression Regulation, Developmental; D051272:Glucose Transporter Type 1 / Q000235:genetics / Q000378:metabolism; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D009028:Morula / Q000166:cytology / Q000378:metabolism*; D011247:Pregnancy; D012333:RNA, Messenger / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D044967:Serum / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000378:metabolism / Q000494:pharmacology
D003470:Culture Media; D051272:Glucose Transporter Type 1; D018840:HSP70 Heat-Shock Proteins; D012333:RNA, Messenger; C081884:heat-shock protein 70.1; D012710:Serum Albumin, Bovine