Role of cytosolic malate dehydrogenase in oocyte maturation and embryo development.
OBJECTIVE
To elucidate the function of cytosolic malate dehydrogenase (Mor2) in oocyte maturation and embryo development using RNA interference (RNAi).


METHODS
Experimental animal study.


METHODS
Research unit of university.


METHODS
Female 4-week-old (C57/BL6) mice.


METHODS
Isolation of immature germinal vesicle (GV) oocytes or fertilized pronucleus (PN) embryos, microinjection of Mor2 double-stranded RNA (dsRNA), and reverse transcription and polymerase chain reaction (RT-PCR) analysis to investigate Mor2-specific messenger RNA (mRNA) knockdown.


METHODS
Relative changes in mRNA levels after microinjection of Mor2 dsRNA and in rates of oocyte maturation and preimplantation embryo development.


RESULTS
Mor2 mRNA mostly was knocked down in germinal vesicle- and metaphase I (MI)-arrested oocytes, compared with metaphase II (MII)-developed oocytes, after microinjection of Mor2 dsRNA and in vitro culture for 16 hours. In vitro oocyte maturation was significantly decreased (34%), compared with noninjected (73.4%) and buffer-injected (67.5%) control groups. The rate of blastocyst development (48.1%) was lower in the Mor2 dsRNA-injected group than in buffer-injected control (88.2%).


CONCLUSIONS
In the present study, the function of Mor2 was analyzed with the aid of RNAi. On the basis of the data obtained, we propose that Mor2 is an essential factor for oocyte maturation and embryo development in mouse.
Yoon
Se-Jin
SJ
CHA Research Institute, Fertility Center, CHA General Hospital, Seoul, Korea.
Koo
Deog-Bon
DB
Park
Jung Sun
JS
Choi
Kyung-Hee
KH
Han
Yong-Mahn
YM
Lee
Kyung-Ah
KA
D000818:Animals; D003600:Cytosol / Q000201:enzymology*; D004247:DNA / Q000008:administration & dosage / Q000494:pharmacology; D047108:Embryonic Development / Q000502:physiology*; D005260:Female; D007267:Injections; D008291:Malate Dehydrogenase / Q000235:genetics / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D009865:Oocytes / Q000502:physiology*; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
D012333:RNA, Messenger; D004247:DNA; D008291:Malate Dehydrogenase; C514238:Mor2 protein, mouse