Effects of oocyte quality on development and transcriptional activity in early bovine embryos.
The objective of this study was to evaluate the effects of oocyte quality on in vitro development and the level of transcriptional activity in early bovine embryos. Cumulus-oocyte complexes (COC) were divided into six classes based on their cumulus investment and on the texture of the ooplasm. Embryos originating from oocytes with more than five layers of cumulus cells and with slight expansion of the cumulus and/or granulation in the ooplasm (class II) developed to the blastocyst stage as frequently as embryos originating from oocytes of class I which showed no signs of atresia (13.9 and 13.7% for classes I and II, respectively). Oocytes with fewer than five layers of cumulus cells and homogeneous ooplasm (class III) had lower cleavage (63.1%) than oocytes with more than five layers of cumulus cells (77.2 and 83.6% for classes I and II, respectively); however, development to the blastocyst stage was similar (12.7%). More advanced atresia such as the presence of granulations in oocytes with less than five layers of cumulus cells (class IV), the absence of cumulus (class V), or the presence of expanded cumulus with dark clumps (class VI) reduced cleavage (57.4, 35.9 and 56.3% for classes IV-VI, respectively) and blastocyst formation (3.5, 0.5 and 1.9% for classes IV-VI, respectively). We examined the effects of oocyte quality on the level of transcriptional activity in in vitro produced embryos at the 2-, 4-, 8-, 16-cell stage and in embryos remaining at the 8-cell stage while the majority had progressed to the 16-cell stage (8-cell delayed embryos) by labeling with 3H-uridine followed by RNA precipitation and scintillation counting. For each developmental stage, there was no significant effect of oocyte class on uptake and incorporation of 3H-uridine into embryos, with the exception of uptake at the 8-cell stage which was higher (P<0.05) in embryos from class V-VI oocytes. Labeled uridine uptake (in embryos from classes I-II and III oocytes) and incorporation (in embryos from oocytes of all classes) increased significantly at the 16-cell stage compared to earlier stages. Eight-cell delayed embryos originating from classes I to IV oocytes incorporated significantly more 3H-uridine than normally developing 8-cell embryos. In conclusion, these results expand on previous work showing that oocytes with early signs of atresia have good development potential. No differences in transcriptional activity in embryos originating from different classes were detected. However, the results obtained with 8-cell delayed embryos indicated that transcriptional activity was determined by the interval after fertilization rather than the number of cell cycles.
Bilodeau-Goeseels
Sylvie
S
Livestock Sciences Section, Agriculture and Agri-Food Canada Research Centre, Lethbridge, Alta, Canada. goeseelss@em.agr.ca
Panich
Paul
P
D000818:Animals; D001755:Blastocyst / Q000502:physiology; D002417:Cattle / Q000196:embryology*; D046508:Culture Techniques; D005314:Embryonic and Fetal Development; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary; D009865:Oocytes / Q000145:classification / Q000502:physiology*; D006080:Ovarian Follicle / Q000166:cytology; D012313:RNA / Q000096:biosynthesis; D014158:Transcription, Genetic; D014316:Tritium; D014529:Uridine / Q000378:metabolism
D014316:Tritium; D012313:RNA; D014529:Uridine