Browse > Article

Characterization and Functional Analysis of Obox4 during Oocyte Maturation by RNA Interference  

Lee, Hyun-Seo (Graduate School of Lift Science and Biotechnology, Pochon CHA University College of Medicine)
Lee, Kyung-Ah (Graduate School of Lift Science and Biotechnology, Pochon CHA University College of Medicine)
Publication Information
Clinical and Experimental Reproductive Medicine / v.34, no.4, 2007 , pp. 293-303 More about this Journal
Abstract
Objective: Previously, we identified differentially expressed genes between GV and MII stage mouse oocytes using ACP technology. When we study one of GV selective genes, Obox family, we found Obox4 mRNA expression in ovaries that has been reported as expressed exclusively in testis. Therefore, this study was conducted for characterization and functional analysis for Obox4. Methods: Expression of Obox4 mRNA was examined in gonads and oocytes by RT-PCR. To determine the role of Obox4 in oocyte maturation, Obox4 dsRNA was microinjected into the cytoplasm of GV oocytes followed by 16 h of incubation in the plain medium or by 24 h of incubation in the medium containing IBMX. After RNAi, phenotypes and maturation rates were observed, change in mRNA expression was evaluated, and chromosomal status was confirmed by orcein staining. Results: Obox4 has minimal expression in the ovary compared to that of the other family members. When oocytes were cultured for 16 h in M16 medium after RNAi, maturation rate was not changed significantly, compared with that of non-injected or buffer-injected control oocytes. Surprisingly, however, when oocytes were cultured for 24 h in M16 containing IBMX, in which oocytes were supposed to arrest at GV stage, Obox4 RNAi oocytes were advanced to MI and MII. Spindle structure was disappeared and the chromosomes were condensed in the oocytes after Obox4 RNAi. Conclusions: This is the first report on the expression of Obox4 in the ovary and oocytes. Results of the study suggest that Obox4 plays a crucial role in spindle formation and chromosome segregation during meiosis in oocytes. In addition, Obox4 may play an important role in cAMP-dependent signal cascades of GV-arrest in mouse oocytes.
Keywords
ACP-PCR; Obox family; Mouse; Oocyte maturation; RNAi;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Barnes FL, Sirad MA. Oocyte maturation. Semin Reprod Med 2000; 18: 123-31   DOI   ScienceOn
2 Cheng WC, Li HMH, Yeh TJ, Li H. Mice lacking the obox6 homeobox gene undergo normal early embryonic development and are fertile. Dev Dyn 2007; 236: 2636-42   DOI   ScienceOn
3 Elbashir SM, Lendeckel W, Tuschl T. RNA interference is mediated by 21- and 22-nucleotide RNAs. Genes Dev 2001; 15: 188-200   DOI   ScienceOn
4 Hammond SM, Bernstein E, Beach D, Hannon GJ. An RNAdirected nuclease mediates post-trasncriptional gene silencing in Drosophila cells. Nature 2000 16; 404: 293-6   DOI
5 Svoboda P, Stein P, Hayashi H, Schultz RM. Selective reduction of dormant maternal mRNA in mouse oocytes by RNA interference. Dev 2002; 127: 4147-56
6 Michaelis C, Ciosk R, Kim N. Cohesins: Chromosomal proteins that prevent premature separation of sister chromatids. Cell 1997; 91: 35-45   DOI   ScienceOn
7 Hutvagner G, Zamore PD. RNAi: nature abhors a double-strand. Curr Opin Genetics Dev 2002; 12: 225-32   DOI   ScienceOn
8 Siard MA. Resumption of meiosis: mechanism involved in meiotic progression and its relation with developmental competence. Theriogenology 2001; 55: 1241-54   DOI   PUBMED   ScienceOn
9 Jones S, Sgouros J. The cohesion complex: sequence homologies, interaction networks and shared motifs. Genome Biol 2001; 2: 0009.1-0009.12
10 Rajkovic A, Van C, Van W, Klysik M, Matzuk MM. Obox, a family of homeobox genes preferentially expressed in germ cells. Genomics 2002; 79: 711-7   DOI   ScienceOn
11 Mongillo M, Zaccolo M. A complex phosphodiesterase system controls -adenoreceptor signaling in cardiomyocytes. Biochem Soc Trans 2006; 34: 510-11   DOI   ScienceOn
12 Richard FJ, Tsafriri A, Conti M. Role of phosphodiesterase type 3A in rat oocyte maturation. Biol Reprod 2001; 65: 1444-51   DOI   ScienceOn
13 Thomas FH, Vanderhyden BC. Oocyte-granulosa cell interactions during mouse follicular development: regulation of kit ligand expression and its role in oocyte growth. Reprod Bio Endocrinol 2006; 4: 19   DOI   ScienceOn
14 Fire A. RNA-triggered gene silencing. Trends Genet 1999; 15: 358-63   DOI   ScienceOn
15 Gil J, Esteban M. Induction of apoptosis by the dsRNAdependent protein kinase (PKR): mechanism of action. Apoptosis 2000; 5: 107-14   DOI   ScienceOn
16 Chen J, Hudson E, Chi MM, Chang AS, Moley KH, Hardie DG, et al. AMPK regulation of mouse oocyte meiotic resumption in vitro. Dev Biol 2006; 291: 227-38   DOI   ScienceOn
17 Rajkovic A, Pangas SA, Ballow D, Suzumori N, Matzuk MM. NOBOX deficiency disrupts early folliculogenesis and oocytespecific gene expression. Science 2004; 30: 1157-9
18 Wianny F, Zemicka-Goetz M. Specific interference with gene function by double-stranded RNA in early mouse development. Nat Cell Bio 2000; 2: 70-5   DOI   ScienceOn
19 Yoon SJ, Chung HM, Cha KY, Kim NH, Lee KA. Identification of differential gene expression in germinal vesicle vs. metaphase II mouse oocytes by using annealing control primers. Fertil Steril 2005; 83: 1293-6
20 Bernstein E, Caudy AA, Hammond SM, Hannon GJ. Role for a bidentate ribonuclease in the initiation step of RNA interference. Nature 2001; 409: 363-6   DOI   ScienceOn