Messenger ribonucleic acid kinetics in human oocytes--effects of in vitro culture and nuclear maturational status.
OBJECTIVE
To study the effects of different nuclear maturational status (prophase I [PI] versus metaphase II [MII]) and in vitro culture on the kinetics of maternal messenger ribonucleic acid (mRNA) in human oocytes.


METHODS
Molecular biology on excess oocytes obtained from our clinical IVF program.


METHODS
The oocytes, classified as either PI or MII at collection, were used as such or cultured in vitro for an additional 24 hours. The relative levels of c-mos and cyclin-B1 were measured using semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR).


RESULTS
The mean levels of c-mos and cyclin-B1 transcripts were indistinguishable between the PI, MII, PI oocytes matured in vitro, PI oocytes failing to mature, and MII oocytes cultured for additional 24 hours. The variability in the levels of these transcripts increased during the vitro culture.


CONCLUSIONS
The level of c-mos and cyclin-B1 transcripts were not different in PI versus MII oocytes, therefore, differences seen in the clinical outcome of PI and MII oocytes may be unrelated to levels of these gene products. C-mos and cyclin B1 mRNA were maintained in vitro, thus degradation of maternal RNA is not activated in excess during the 24-hour culture.
Heikinheimo
O
O
Jones Institute for Reproductive Medicine, Department of Obstetrics and Gynecology, Eastern Virginia Medical School, Norfolk 23507, USA.
Toner
J P
JP
Lanzendorf
S E
SE
Billeter
M
M
Veeck
L L
LL
Gibbons
W E
WE
D001483:Base Sequence; D002467:Cell Nucleus / Q000502:physiology*; D004396:Coloring Agents; D046508:Culture Techniques; D019926:Cyclin B; D056744:Cyclin B1; D016213:Cyclins / Q000235:genetics / Q000378:metabolism; D004996:Ethidium; D005260:Female; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D009865:Oocytes / Q000378:metabolism* / Q000648:ultrastructure; D016133:Polymerase Chain Reaction; D017231:Proto-Oncogene Proteins c-mos / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*
C534699:CCNB1 protein, human; D004396:Coloring Agents; D019926:Cyclin B; D056744:Cyclin B1; D016213:Cyclins; D012333:RNA, Messenger; D017231:Proto-Oncogene Proteins c-mos; D004996:Ethidium