Developmental competence of heifer oocytes selected using the brilliant cresyl blue (BCB) test.
The aim of this study was to evaluate the usefulness of the brilliant cresyl blue (BCB) test in the selection of more competent heifer oocytes for in vitro embryo production (IVEP). IVEP from selected BCB heifer oocytes was compared to IVEP from morphologically selected heifer (control group) and cow oocytes. BCB staining determines the activity of glucose-6-phosphate dehydrogenase (G6PD), an enzyme synthesized in growing oocytes but with less activity in grown oocytes. Six hundred and fifty seven heifer cumulus-oocyte complexes (COC) were classified morphologically as Grade 1-3 and exposed to 26 microM of BCB and classified as: blue (or grown) oocytes (BCB+) or unstained oocytes or growing oocytes (BCB-). Grade 1-3 heifer oocytes showed significantly different percentages of BCB+ oocytes (78.6, 66.2, and 51.1%, respectively; P<0.05). The diameter of BCB+ oocytes was significantly higher than BCB- oocytes (152.6+/-5.8 microm and 147+/-5.9 microm, respectively; P<0.001). The percentage of BCB+ oocytes reaching the blastocyst stage was significantly higher than those of BCB- and control heifer oocytes (12.3, 1.6, and 5.2%, respectively; P<0.05), but lower than those of cow oocytes (30.0%; P<0.05). In conclusion, heifer oocytes selected by the BCB test (BCB+) are larger and more competent for IVEP than control heifer oocytes. However, fewer heifer oocytes selected using the BCB test develop to blastocyst stage compared to cow oocytes.
Pujol
Marc
M
Departament de Ciència Animal i dels Aliments, Facultat de Veterinària, Universitat Autònoma de Barcelona, Bellaterra, Barcelona 08193, Spain.
López-Béjar
Manel
M
Paramio
Maria Teresa
MT
D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000502:physiology; D002417:Cattle; D048429:Cell Size; D004396:Coloring Agents; D046508:Culture Techniques; D005314:Embryonic and Fetal Development; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary; D005954:Glucosephosphate Dehydrogenase / Q000032:analysis; D009865:Oocytes / Q000166:cytology / Q000201:enzymology / Q000502:physiology*; D010078:Oxazines; D020858:Tissue and Organ Harvesting / Q000379:methods / Q000662:veterinary*
D004396:Coloring Agents; D010078:Oxazines; C006797:Brilliant Cresyl Blue; D005954:Glucosephosphate Dehydrogenase