Selection of ovine oocytes by brilliant cresyl blue staining.
Sheep oocytes derived from the ovaries collected from the slaughterhouse are often used for research on in vitro embryo production, animal cloning, transgenesis, embryonic stem cells, and other embryo biotechnology aspects. Improving the in vitro culture efficiency of oocytes can provide more materials for similar studies. Generally, determination of oocyte quality is mostly based on the layers of cumulus cells and cytoplasm or cytoplasm uniformity and colors. This requires considerable experience to better identify oocyte quality because of the intense subjectivity involved (Gordon (2003), Madison et al. (1992) and De Loos et al. (1992)). BCB staining is a function of glucose-6-phosphate dehydrogenase (G6PD) activity, an enzyme synthesized in developing oocytes, which decreases in activity with maturation. Therefore, unstained oocytes (BCB-) are high in G6PD activity, while the less mature oocytes stains are deep blue (BCB+) due to insuffcient G6PD activity to decolorize the BCB dye.
Key Laboratory of Animal Biotechnology of Xinjiang and Key Laboratory of Grass Livestock Reproduction and Breed Biotechnology, Ministry of Agriculture, Animal Science Academy of Xinjiang Uygur Autonomous Region, Urumqi 830000, China.
Liqin
L
Wang
Jiapeng
J
Lin
Juncheng
J
Huang
Jing
J
Wang
Yuncheng
Y
Zhao
Tong
T
Chen
D000704:Analysis of Variance; D000818:Animals; D018929:Cell Culture Techniques; D048429:Cell Size / Q000187:drug effects; D002478:Cells, Cultured; D004396:Coloring Agents / Q000737:chemistry*; D004527:Egg Proteins / Q000235:genetics / Q000378:metabolism; D005260:Female; D005954:Glucosephosphate Dehydrogenase / Q000235:genetics / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism; D009865:Oocytes / Q000737:chemistry / Q000166:cytology* / Q000378:metabolism; D010053:Ovary / Q000166:cytology / Q000378:metabolism; D010078:Oxazines / Q000737:chemistry*; D016133:Polymerase Chain Reaction; D012756:Sheep; D013194:Staining and Labeling / Q000379:methods
D004396:Coloring Agents; D004527:Egg Proteins; D010078:Oxazines; C006797:Brilliant Cresyl Blue; D005954:Glucosephosphate Dehydrogenase; D005978:Glutathione