|Budget Amount *help
¥5,500,000 (Direct Cost : ¥5,500,000)
Fiscal Year 2000 : ¥1,400,000 (Direct Cost : ¥1,400,000)
Fiscal Year 1999 : ¥1,700,000 (Direct Cost : ¥1,700,000)
Fiscal Year 1998 : ¥2,400,000 (Direct Cost : ¥2,400,000)
This grant has been a great help for us to set up and carry out those projects listed below. Already, some results has been published, and the other results are close to be published.
1) Research on development of mice ovary.
1-1. Analysis of anomaly in development of follicles and abnormal proliferation of follicular granulosa cell in gdf-9 -/- mice ovarian development :
To further analyze the defect in ovarian follicular development and abnormal proliferation of ovarian granulosa cells of gdf-9-/-mice, we studied the expression pattern of various gene suspected to be directed by gdf-9 and have more direct effect giving rise to the lesion in ovarian follicular development. We showed that the level of the expression of c-kit ligand and hyaluronic acid synthetase might be suppressed in those mutant mice.
1-2. Generation of transgenic mice harboring DAX-1-cre :
We generated the transgenic mice harboring DAX-1-cre which can facilitate stroma cell specific excision of the target genes such as A
LK3 , by cre/LoxP mediated recombination in ovaries and testes in mice.
2) Analysis of its vivo function of otr and ot gene system by generating gene knockout mice.
2-1. Generation of otr gene conditional deficient (knockout) mice :
We generated mice whose otr gene was substituted with manipulated otr gene, surrounded by two LoxP sequences (floxed otr).
Progeny mice born from ^<flx>otr/^<flx>otr female mice died within 36 hr after birth. This phenotype was suspected to be caused by decreasing level in expression of otr gene in the mutant mice. The quantitation of the level of otr gene expression in these mutant mice were now on going.
2-2. Generation of otr gene (conventional knockout) mice :
By crossing ^<flx>otr/wt mouse with CAG-cre mice known as general deleter, we obtained conventional otr gene knockout mice. On the contrary to the first expectation, male and female otr-/- mice were so far viable, looked healthy and showed no obvious defect in their appearance. However, viability of progeny mice from the otr heterozygous mice were tend to be lower than those from WT parents.
2-3. The system of ot/otr in vivo has remarkable effect on neuronal activity in brain including behavior, learning and memory. After obtaining otr-/- mice at January 2001, we began collaborative research with Dr. Thomas Insel, the director of Center of Behavior and Neuroscience, Emory University, GA, USA.The major purpose of this collaboration is to analyze the abnormal behavior in otr-/- mice, and to use these mutant mice as a material to analyze the function of otr gene ectopically expressed by virus vectors in mice brain.
3) Functional Analysis of MTA-1 gene.
3-1 MTA-1 gene is showing strong correlation with metastasis and malignancy of cancer cell derived from reproductive organs such as ovary and mammary gland. We reported the effect of MTA-1 gene expression on cell proliferation, on the cell's metastatic ability and on the transcriptional activity of GATA enhancer element. Less