Altered composition of the cumulus-oocyte complex matrix during in vitro maturation of oocytes.
BACKGROUND
In vitro maturation (IVM) of mammalian oocytes has potential health benefits for patients undergoing assisted reproduction as an alternative to gonadotrophin treatment. This procedure is also useful for studying the process of oocyte and early embryo development. However, oocytes undergoing IVM have much lower competence than in vivo matured oocytes. Efforts to optimize IVM success have focused on replicating in vivo timing, hormonal milieu and cumulus cell responses associated with maturing oocytes. We have previously identified two extracellular matrix proteins, the protease Adamts1 and hyaluronan-binding proteoglycan Versican, produced by mural granulosa cells that selectively incorporate into the periovulatory cumulus-oocyte complex (COC).


METHODS
Murine COC were cultured in the presence of epidermal growth factor and/or FSH. mRNA and protein were measured by real time PCR and Western blot and compared to in vivo derived COC.


RESULTS
COCs from mice that underwent IVM for 6 or 20 h in the presence of epidermal growth factor, FSH or in combination had a > 10-fold reduction in mRNA (P < 0.05) for Adamts1 and Vcan when compared with in vivo matured COCs. Hyaluronan synthase 2 expression was up-regulated up to 8-fold (P < 0.05) over the unstimulated control, demonstrating successful induction of cumulus gene expression by the IVM conditions. While in vivo matured COCs showed abundant levels of these proteins, COCs that underwent IVM had neither detectable Adamts1, nor intact or Adamts1-cleaved Vcan. Human cumulus and granulosa cells matured in vivo contained abundant mRNA for Adamts1 and Vcan, demonstrating the potential relevance to human IVM.


CONCLUSIONS
These results indicate that extensively altered COC matrix composition is present during IVM and may contribute to the observed poorer competence of the derived oocytes.
Dunning
Kylie R
KR
Discipline of Obstetrics and Gynaecology, Research Centre for Reproductive Health, The University of Adelaide, Adelaide, SA, Australia.
Lane
Michelle
M
Brown
Hannah M
HM
Yeo
Christine
C
Robker
Rebecca L
RL
Russell
Darryl L
DL
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D006063:Chorionic Gonadotropin / Q000378:metabolism; D054885:Cumulus Cells / Q000166:cytology*; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods*; D005640:Follicle Stimulating Hormone / Q000378:metabolism; D005786:Gene Expression Regulation; D006107:Granulosa Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D008954:Models, Biological; D009865:Oocytes / Q000166:cytology* / Q000378:metabolism; D010053:Ovary / Q000378:metabolism / Q000473:pathology
D006063:Chorionic Gonadotropin; D005640:Follicle Stimulating Hormone