Effects of oocyte maturation regimen on the relative abundance of gene transcripts in bovine blastocysts derived in vitro or in vivo.
Bovine embryos produced in vitro differ substantially from embryos produced in vivo in the mRNA expression patterns of genes important for development. Several factors in the in vitro production systems have profound effects on embryonic mRNA expression patterns. The effects of the type of maturation on the expression pattern of genes important for development in blastocysts produced in vitro have not yet been investigated. The aim of the present study was to investigate the effects of various maturational protocols on the relative abundance of a panel of six marker genes, indicative of compaction and cavitation, metabolism, stress susceptibility and RNA processing, in bovine blastocysts produced in vitro. Four groups of blastocysts were analysed by a sensitive semi-quantitative RT-PCR assay. Blastocysts were produced in vitro from oocytes of different origin from: (1) 3-8 mm follicles; (2) preovulatory follicles before the LH surge; and (3) preovulatory follicles 24 h after the LH surge. The first two groups were matured in vitro, whereas the third group had undergone maturation in vivo. A fourth group comprised blastocysts developed entirely in vivo. Expression of glucose transporter 1 was significantly (P < 0.05) higher, and expression of desmocollin 2 and plakophilin tended to be higher (P < 0.1) for in vivo (group 4) compared with in vitro blastocysts (group 1), whereas no differences were found for heat shock protein 70.1, E-cadherin and poly(A) polymerase. Expression of the six transcripts did not differ among blastocysts produced in vitro from oocytes of groups 1, 2 and 3. Results indicate that alterations in the relative abundance of these transcripts in blastocysts produced in vitro cannot primarily be attributed to the origin of the oocyte, but are likely to have been induced by post-maturation or fertilization culture conditions.
Department of Farm Animal Health, Faculty of Veterinary Medicine, University of Utrecht, Yalelaan 7, 3584 CL Utrecht, The Netherlands. H.Knijn@vet.uu.nl
H M
HM
Knijn
C
C
Wrenzycki
P J M
PJ
Hendriksen
P L A M
PL
Vos
D
D
Herrmann
G C
GC
van der Weijden
H
H
Niemann
S J
SJ
Dieleman
D000818:Animals; D001755:Blastocyst / Q000166:cytology*; D002417:Cattle / Q000196:embryology*; D005314:Embryonic and Fetal Development / Q000235:genetics; D005260:Female; D005307:Fertilization in Vitro; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D005819:Genetic Markers; D066298:In Vitro Techniques; D007986:Luteinizing Hormone / Q000097:blood; D009865:Oocytes / Q000166:cytology*; D016133:Polymerase Chain Reaction / Q000379:methods; D011247:Pregnancy; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013480:Superovulation; D014158:Transcription, Genetic
D005819:Genetic Markers; D007986:Luteinizing Hormone