Establishment of ovarian reconstruction system in culture for functional genomic analysis.
In a previous study, we reported a list of genes expressed differentially in primordial and primary follicles [Park et al., Fertil. Steril., 83, 410-418 (2005)]. An innovative experimental system is required to evaluate the functions of these genes in folliculogenesis, particularly primordial-primary follicle transition. In this study, ovarian tissues were dissociated, and isolated cells were transfected using small interfering RNAs (siRNAs) for disrupting a specific gene, followed by ovarian reconstruction via calcium alginate encapsulation. The effects of RNA interference (RNAi) on follicular development were evaluated by a histological observation of the reconstructed ovarian tissue. Interestingly, follicular formation and development showed differences between control and experimental groups. Thus, even though this system includes some problems that need to be solved, ovarian reconstruction following the modification of gene expression of individual ovarian component cells could lead the way to methods of studying molecular mechanisms of primordial-primary follicle transition.
Park
Chang-Eun
CE
Graduate School of Life Science and Biotechnology, Pochon CHA University College of Medicine, CHA Research Institute, Fertility Center, CHA General Hospital, 606-13 Yeoksam-1-dong, Gangnam-gu, Seoul 135-081, South Korea.
Lee
Dongryul
D
Kim
Kyeoung-Hwa
KH
Lee
Kyung-Ah
KA
D000818:Animals; D018797:Cell Cycle Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000502:physiology; D002478:Cells, Cultured; D005260:Female; D051379:Mice; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000502:physiology; D009865:Oocytes / Q000502:physiology; D006080:Ovarian Follicle / Q000502:physiology*; D016133:Polymerase Chain Reaction; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000502:physiology; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000494:pharmacology; D014162:Transfection
D018797:Cell Cycle Proteins; D009687:Nuclear Proteins; D034741:RNA, Small Interfering; D011505:Protein-Tyrosine Kinases; C499348:Wee1 protein, mouse