Premium
Cyclin B 1 expression in meiotically competent and incompetent goat oocytes
Author(s) -
Hue Isabelle,
Dedieu Thierry,
Huneau Denise,
Ruffini Sylvie,
Gall Laurence,
Crozet Nicole
Publication year - 1997
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/(sici)1098-2795(199706)47:2<222::aid-mrd14>3.0.co;2-1
Subject(s) - biology , cyclin b1 , cyclin b , cyclin , microbiology and biotechnology , cyclin a , complementary dna , cyclin d , cyclin a2 , cyclin d2 , messenger rna , cyclin e , cyclin e1 , cell cycle , gene , genetics , cyclin dependent kinase 1
To start determining the nature of meiotic incompetence in goat oocytes, we have examined the expression of one of the potential pre‐MPF subunits: the cyclin B 1 . We have been isolating a small DNA probe encoding the goat cyclin B 1 box to analyze the expression of the cyclin B 1 gene in competent and incompetent goat oocytes. This probe was easily obtained by polymerase‐chain‐reaction (PCR) on reverse‐transcribed mRNA from granulosa cells, using cyclin B specific primers derived from a bovine cDNA. The transcript corresponding to cyclin B 1 in goat granulosa cells is 1.8 kb. In situ hybridization analysis indicated that competent and incompetent oocytes contained cyclin B 1 mRNA, but also that active cyclin B 1 mRNA synthesis occured at the end of the growth phase, e.g., when oocytes progressed in the acquisition of meoitic competence. Western blot analysis, performed with a monoclonal anticyclin B 1 antibody, revealed in competent and incompetent oocytes a polypeptide of 65kDa corresponding to the goat cyclin B 1 protein. This pattern of cyclin B 1 expression further suggested that meiotic incompetence in goat oocytes could not be primarily correlated with a lack of cyclin B 1 protein as potential pre‐MPF subunit, but to a limiting amount of this protein. Mol. Reprod. Dev. 47:222–228, 1997. © 1997 Wiley‐Liss, Inc.