[Growth differentiation factor-9 gene expression in in vitro cultured oocytes in mice].
OBJECTIVE
To explore the relation between oocyte maturation and growth differentiation factor-9 (GDF-9) gene expression.


METHODS
Ovariectomy was performed in 50 Kunming female mice of 10 days old, and the preantral follicles were isolated from the ovaries and cultured in medium drops for 12 days. Oocytes and somatic cells were mechanically isolated. The oocytes cultured in vitro for 2, 4, 6, 8, 10, and 12 days constituted the in vitro cultured group and the oocytes obtained from female mice of 12, 14, 16, 18, 20, and 22 days old served as the in vivo group. Semi-quantitative RT-PCR and agar gel electrophoresis were performed to quantify GDF-9 gene expression in each oocyte.


RESULTS
Follicle survival, antrum formation and maturation rate was 89.5%, 51.8% and 56.6% in the in vitro cultured follicles, respectively. GDF-9 gene expression on days 2, 4, 6, 8, 10, and 12 in in vitro cultured oocytes was 0.83-/+0.08, 0.52-/+0.09, 0.45-/+0.13, 0.49-/+0.09, 0.49-/+0.09, and 0.68-/+0.08, respectively; GDF-9 gene expression in in vivo grown oocytes of 12, 14, 16, 18, 20, and 22 days were 0.64-/+0.35, 0.48-/+0.10, 0.52-/+0.10, 0.66-/+0.08, 0.72-/+0.09, and 0.91-/+0.11, respectively. Between days 8 and 12, GDF-9 gene expression in in vitro cultured oocyte was significantly lower than that in in vivo grown oocytes (P<0.05).


CONCLUSIONS
MII oocytes can be obtained from in vitro culture of the preantral follicles. GDF-9 gene expression in the oocytes varies with their growth stages. Between days 8 and 12 of in vitro culture, GDF-9 gene expression in the cultured oocytes is different from that in in vivo grown oocytes.
Peng
Yu-hong
YH
Department of Obstetrics and Gynecology, Wuhan General Hospital of Guangzhou Command, Wuhan 430070, China. pyhlucy@yahoo.com.cn
Zhuang
Guang-lun
GL
Zhou
Can-quan
CQ
D000818:Animals; D000831:Animals, Newborn; D055430:Bone Morphogenetic Protein 15; D002470:Cell Survival; D002478:Cells, Cultured; D004587:Electrophoresis, Agar Gel; D005260:Female; D015870:Gene Expression; D055429:Growth Differentiation Factor 9; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics*; D051379:Mice; D009865:Oocytes / Q000166:cytology / Q000254:growth & development / Q000378:metabolism*; D006080:Ovarian Follicle / Q000166:cytology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors
C525902:Bmp15 protein, mouse; D055430:Bone Morphogenetic Protein 15; C525898:Gdf9 protein, mouse; D055429:Growth Differentiation Factor 9; D036341:Intercellular Signaling Peptides and Proteins