Impact of in vitro production techniques on the expression of X-linked genes in bovine (bos taurus) oocytes and pre-attachment embryos

• Published in: Molecular reproduction and development 2007-02, Vol.74 (2), p.144-153
• Main Authors: Nino-Soto, Maria I., Basrur, Parvathi K., King, W. Allan
• Format: Article
• Language: English
• Subjects: Animals; Base Sequence; Biological and medical sciences; Blastocyst - physiology; Bos taurus; Cattle; DNA Primers; DNA, Complementary - genetics; Early stages. Segmentation. Gastrulation. Neurulation; Embryology: invertebrates and vertebrates. Teratology; Embryonic Development; Fertilization in Vitro - veterinary; Fundamental and applied biological sciences. Psychology; gene expression; Morula - physiology; Oocytes - physiology; Polymerase Chain Reaction; reproductive biotechnology; RNA - genetics; RNA - isolation & purification; Transcription, Genetic; X Chromosome; XIST
• ISSN: 1040-452X; 1098-2795
• DOI: 10.1002/mrd.20575

Our previous studies showed that expression patterns of X‐linked genes in cultured cells are different from those of their tissues of origin. This investigation analyses the transcription pattern of the X‐linked genes BIRC4, GAB3, MECP2, RPS4X, SLC25A6, and XIST in bovine in vitro matured oocytes and in vitro fertilized embryos, and their in vivo counterparts. In vitro‐derived pools of mature oocytes and pre‐attachment embryos were obtained by: (a) TCM‐199/serum with bovine oviductal epithelial cells as co‐culture, and (b) synthetic oviductal fluid/BSA. Pools of in vivo‐derived morulae and blastocysts were provided by a commercial embryo transfer operation. Total RNA was extracted for quantification of gene‐specific transcript levels using real‐time quantitative PCR. Statistical analysis was performed using a mixed model factorial ANOVA with α = 0.05. The effect of the in vitro environmental conditions on X‐linked gene transcription was most evident during the fourth cell cycle, at the period of activation of the embryonic genome, and seemed to be less pronounced at later developmental stages, with the exception of BIRC4. The levels of X‐linked genes transcripts in in vivo‐derived embryos were lower relative to their in vitro counterparts for all genes tested. Finally, the pattern of expression of XIST in bovine oocytes and embryos was similar to that reported in humans. These results highlight the possibility that X‐linked gene expression analysis is a useful tool to monitor the impact of reproductive biotechnologies on the developmental potential of embryos and aid in their improvement. Mol. Reprod. Dev. © 2006 Wiley‐Liss, Inc.