1997 Fiscal Year Final Research Report Summary
Research on regulation mechanisms of development and differentiation of mouse primordial germ cells
| Project/Area Number |
08458241
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Developmental biology
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| Research Institution | National Institute of Genetics |
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Principal Investigator |
NAKATSUJI Norio National Institute of Genetics, Mammalian Development Laboratory, Professor, 系統生物研究センター, 教授 (80237312)
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| Co-Investigator(Kenkyū-buntansha) |
SHIRAYOSHI Yasuaki National Institute of Genetics, Mammalian Development Laboratory, Assistant prof, 医学部, 助教授 (90249946)
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| Project Period (FY) |
1996 – 1997
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| Keywords | mouse / primordial germ cells / sex differentiation / meiosis / testis / ovary / cell culture |
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| Research Abstract |
Mouse primordial germ cells appear at the base of allantois in the posterior extraembryonic region, migrate to the genital ridges, and differentiate into oocytes or prospermatogonia in the fetal ovary or testis. We carried out the following experiments to study mechanisms in the development and differentiation of fetal germ cells.
There have been no culture system of fetal germ cells after arrival at gonads, because they go into apoptosis and disappear in usual culture conditions. We developed novel methods of sexing embryos, isolation of germ cells with magnetic beads, and detection of meiotic cells with specific antibodies. As a result, we detected many germ cells entering into meiotic prophase in our culture system.
Mammalian sex-determination and differentiation is controlled by several genes, such as Sry, Sox-9, Dax-1 and Ad4BP/SF-1, but their upstream and downstream genes are largely unknown. In order to identify these genes involved in the sex-differentiation, we carried out the differential hybridization of the mouse embryonic gonad cDNA library using presumably male-specific probes. We identified nephgonadin, which encodes a basic helix-loop-helix motif. The earliest expression of nephgonadin was observed at 8.5 days post coitum (dpc) in the intermediate mesodermal tissues of anterior and posterior parts of the mouse embryo. The posterior expression continued until this region forms the urogenital ridge. From 13.5 dpc to 2 weeks postnatal, nephgonadin was expressed at higher levels in the male than female gonad. In adults, however, expression of nephgonadin was drastically decreased in the testis, while it was increased in the ovary. These sex- and stage-dependent expression patterns are similar to that of Ad4BP/SF-1, which is important for the development and sex-differentiation of the gonad.
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[Publications] Koshimizu, U., Taga, T., Watanabe, M., Saito, M., Shirayoshi, Y., Kishimoto, T.and Nakatsuji, N.: "Functional requirement of gp130-mediated signaling for growth and survival of mouse primordial germ cells in vitro and derivation of embryonic germ (EG) cells" Development. 122. 1235-1242 (1996)
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[Publications] Tamura, M., Kanno, Y., Chuma, S., Wakana, S., Saito, T., Shirayoshi, Y.and Nakatsuji, N.: "A basic helix-loop-helix transcription factor gene, nephgonadin, isolated by differential screening of the male and female fetal gonads shows a sex- and stage-dependent expression pattern in the gonad development." (Submitted for publication.).
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