title: A combination of FSH and dibutyryl cyclic AMP promote growth and acquisition of meiotic competence of oocytes from early porcine antral follicles.
abstract: Growing porcine oocytes from early antral follicles (1.2-1.5 mm in diameter) do not mature to metaphase II (MII, 4%) under culture conditions which supported maturation (MII, 95%) of fully grown oocytes from large (4-6 mm) antral follicles. We hypothesized that FSH and dbcAMP supported growth and acquisition of meiotic competence. Growing oocytes (113.0 ± 0.4 μm, mean ± SEM) were cultured for 5 d in medium supplemented with 1 mM dbcAMP, 0.01 IU/mL FSH or both; in these media, oocytes reached, 120.5 ± 0.4, 123.5 ± 0.4 and 125.7 ± 0.2 μm, respectively, after 5 d, and then were matured in vitro for 48 h. Oocytes remained enclosed by cumulus cells when cultured with FSH (82%) or both FSH and dbcAMP (80%), but not with dbcAMP alone (0%). Furthermore, oocytes cultured with FSH maintained trans-zonal projections of cumulus cells. Oocytes remained at the GV stage at higher rates when cultured with dbcAMP and FSH (99%), or dbcAMP (97%), than with FSH (64%), or without either (75%). Following in vitro maturation, oocytes reached MII after in vitro growth with dbcAMP (19%), FSH (11%), or both (68%). When oocytes were cultured with both FSH and dbcAMP, activation of Cdc2 and MAP kinases in growing oocytes was similar to fully grown oocytes. In conclusion, growing porcine oocytes grew and acquired meiotic competence in medium supplemented with dbcAMP and FSH; the former maintained oocytes in meiotic arrest, whereas the latter maintained trans-zonal projections of cumulus cells to oocytes during in vitro growth culture.
authors.affiliation: Graduate School of Agricultural Science, Kobe University, Kobe 657-8501, Japan. 064A251A@stu.kobe-u.ac.jp
authors.forename: Ilse Silvia
authors.initials: IS
authors.lastname: Cayo-Colca
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Yamagami
authors.forename: Thuy-Chau
authors.initials: TC
authors.lastname: Phan
authors.forename: Takashi
authors.initials: T
authors.lastname: Miyano
mesh_terms: D000818:Animals; D003994:Bucladesine / Q000494:pharmacology*; D016203:CDC2 Protein Kinase / Q000378:metabolism; D018929:Cell Culture Techniques; D003470:Culture Media; D054885:Cumulus Cells / Q000166:cytology; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005260:Female; D005640:Follicle Stimulating Hormone / Q000494:pharmacology*; D008540:Meiosis / Q000187:drug effects*; D009865:Oocytes / Q000166:cytology / Q000187:drug effects* / Q000254:growth & development; D006080:Ovarian Follicle / Q000166:cytology / Q000187:drug effects*; D013552:Swine / Q000502:physiology*
chemical_list: D003470:Culture Media; D003994:Bucladesine; D005640:Follicle Stimulating Hormone; D016203:CDC2 Protein Kinase; D048049:Extracellular Signal-Regulated MAP Kinases