Alterations in the relative abundance of gene transcripts in preimplantation bovine embryos cultured in medium supplemented with either serum or PVA.
In preimplantation bovine embryos, the relative abundance of various developmentally important gene transcripts was determined by a semi-quantitative RT-PCR assay to analyze the effects of two medium supplements, serum or polyvinyl alcohol (PVA). Development to morula, blastocyst, and hatched blastocyst stages was higher (P < or = 0.05) in medium supplemented with serum than in medium supplemented with PVA. Connexin43 mRNA expression virtually disappeared from the 8-16 cell stage onward, but reappeared in the hatched blastocyst in serum-supplemented medium, whereas it was detected in PVA-derived embryos throughout development. No differences were found for plakophilin mRNA between both culture groups. Desmocollin II mRNA showed a sharp increase at the blastocyst stage in both groups with a higher transcription level in PVA-generated embryos. A significant difference in desmocollin III transcripts was detectable at the 8-16-cell stage between serum- and PVA-derived embryos. Transcripts for desmoglein 1 and desmocollin I were not detected at any preimplantation stage, irrespective of medium supplementation. The relative abundance of glucose-transporter-1 mRNA was significantly increased at the 8-16-cell stage in embryos produced in medium supplemented with PVA, but not serum. Heat shock protein and poly(A)polymerase mRNA were continuously expressed during preimplantation development in both culture groups. Although poly(A)polymerase mRNA was significantly elevated in PVA- over serum-derived embryos, heat shock protein mRNA expression was significantly enhanced in serum-generated embryos over PVA-derived embryos. Interferon tau mRNA showed a significant increase at the hatched blastocyst stage only in PVA-supplemented medium. These data suggest that alterations in mRNA expression are associated with culture environment. Timing and magnitude of the alterations varied among the different transcripts and were significantly affected by the presence of exogenous protein in a stage-specific manner, predominantly at critical developmental time points.
Wrenzycki
C
C
Department of Biotechnology, Institut für Tierzucht und Tierverhalten (FAL), Mariensee, Neustadt, Germany.
Herrmann
D
D
Carnwath
J W
JW
Niemann
H
H
D000818:Animals; D002417:Cattle; D003470:Culture Media; D047108:Embryonic Development / Q000502:physiology; D005314:Embryonic and Fetal Development; D005260:Female; D018507:Gene Expression Regulation, Developmental; D011142:Polyvinyl Alcohol; D011247:Pregnancy; D012333:RNA, Messenger; D012710:Serum Albumin, Bovine
D003470:Culture Media; D012333:RNA, Messenger; D012710:Serum Albumin, Bovine; D011142:Polyvinyl Alcohol