Factors affecting the developmental competence of mouse oocytes grown in vitro: follicle-stimulating hormone and insulin.
This study was undertaken to test the hypothesis that FSH treatment of cultured oocyte-granulosa cell complexes promotes acquisition of competence to complete preimplantation embryo development. Oocyte-granulosa cell complexes were isolated from the preantral follicles of 12-day-old mice and cultured for 10 days in serum-free medium, supplemented with insulin (5 microgram/ml), transferrin (5 microgram/ml), and selenium (5 ng/ml) and containing a highly potent preparation of FSH (0-5 ng/ml). Oocytes were matured and fertilized in vitro and embryos cultured to determine the frequency of development to the blastocyst stage. There was no effect of FSH on oocyte size, general morphology, or competence to resume meiosis. However, addition of FSH to medium containing insulin had a deleterious effect on the percentage of mature oocytes competent to develop to the blastocyst stage. Deletion of insulin from the medium for culture of oocyte-granulosa cell complexes prevented the deleterious effect of FSH, but FSH still did not promote acquisition of competence to complete preimplantation development. Culture of oocyte-granulosa cell complexes with FSH resulted in elevated expression of LH receptor (LHR) mRNA by granulosa cells and stimulated the production of functional LHRs, whether or not insulin was present. However, FSH-induced expression of LHR mRNA reached a maximum steady-state level by 4 days of culture in the presence of insulin, but this level was not reached until 10 days of culture without insulin. Granulosa cells encompassing growing mouse oocytes in vivo do not express LHR mRNA. Thus, expression of LHR mRNA by granulosa cells closely associated with growing oocytes in vitro indicates inappropriate or ambiguous development. In conclusion, conditions occurring during oocyte growth can have profound detrimental effects on oocyte developmental competence to complete preimplantation development, even when oocyte growth, general morphology, and competence to resume meiosis appear unaffected.
Eppig
J J
JJ
The Jackson Laboratory, Bar Harbor, Maine 04609-1500, USA. jje@jax.org
O'Brien
M J
MJ
Pendola
F L
FL
Watanabe
S
S
D000818:Animals; D001755:Blastocyst / Q000502:physiology; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D005260:Female; D005307:Fertilization in Vitro; D005640:Follicle Stimulating Hormone / Q000494:pharmacology*; D006107:Granulosa Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D007328:Insulin / Q000494:pharmacology*; D051379:Mice; D009865:Oocytes / Q000187:drug effects / Q000254:growth & development*; D012333:RNA, Messenger / Q000378:metabolism; D011974:Receptors, LH / Q000235:genetics; D012643:Selenium / Q000494:pharmacology; D014168:Transferrin / Q000494:pharmacology
D016895:Culture Media, Serum-Free; D007328:Insulin; D012333:RNA, Messenger; D011974:Receptors, LH; D014168:Transferrin; D005640:Follicle Stimulating Hormone; D012643:Selenium